ENZYMATIC REVERSE HYDROLYSIS OF HYDROPHILIC SUBSTRATES - PREPARATION OF AMPHIPHILIC COMPOUNDS

Abstract: The invention relates to methods for the production of amphiphilic products such as esters, sugar-esters, peptide-esters, glycolipids, glycoproteins, lipoproteins, peptides, and phosphates of alcohols, sugars, and nucleosides. The methods promote enzymatically catalyzed reactions between hydrophilic substrates such as glucerol, glucose, amino acids, and nucleosides, and second substrates such as free fatty acids, triglycerides, vinylesters, amino acids, and phosphates. The method is also applied to enzymatic reactions with saccharides and polyalcohols. The hydrophilic substrates are adsorbed to finely divided solid supports such as silica gel, diatomaceous earths, or activated chracoals in order to promote the dispersion of the hydrophilic substrates within hydrophobic substrates and solvents. Hydrophobic solvents such as n-hexane and t-butylmethylether may be included in the reaction mixtures. Reactions are conducted under non-aqueous conditions in order to promote reverse hydrolysis. Methods are provided for the production of isomerically pure 1,3-diglycerides. Further methods are disclosed for the production and specific precipitation of pure 1-monogycerides through the use of a reactor/separator system. Enzymes used in the methods include lipases from M. mihei and P. fluorescens, glycosidases such as beta - galactosidase, proteases such as chymotrypsin, and acid or alkaline phosphatases. Compositions are provided comprising alcohols, carbohydrates, amino acids, or peptides adsorbed onto solid supports such as silica gel.

Description: EN2YMATIC REVERSE HYDROLYSIS OF HYDROPHILIC SUBSTRATES - PREPARATION OF AKPHIPHILIC COMPOUNDS CROSS-REFERENCE This application is a continuation-in-part of co-pending U.S. Patent Application Serial Number 07/654,979 which was filed on February 13, 1991. TECHNICAL FIELD OF THE INVENTION The invention relates to methods for producing amphiphillc compounds through the biocatalyzed reaction of a hydrophilic substrat , adsorbed onto a solid support, with a second substrate, which may be hydrophobic. The invention relates to methods for producing Isomerically pure 1,3-diglycerides and 1-monoglycerides, sugar esters, amino acid esters, peptides, and glycolipids, as well as phosphates of alcohols, carbohydrates, and nucleosides. The invention provides compositions of an alcohol or a carbohydrate adsorbed to a solid support and compositions of a carboxyl-protected amino acid or a carboxyl-protected peptide adsorbed to a solid support in combination with an amino-protected amino acid or an amino-protected peptide adβorbed to the sane support. The invention also provides methods for the selective precipitation of l-monoglyceride products. Riπyr.PfflTKP OF THE INVENTION Many chemicals useful in industry are amphiphilic compounds which can be obtained by esterification or reverse hydrolytic reactions between hydrophilic compounds and second compounds, which may be hydrophobic. Illustrations of such reactions follow. 1) Esterification of a polyalcohol. glycerol + free fatty acid -> aono-g yceridβ + water 2) Esterification of a carbohydrate: glucose + free fatty acid -* sugar-ester + water 3) Esterification of an amino acid: peptide + free tty acid «♦ peptide ester + water The amphiphilic products of such reactions are used as surfactants, e ulsifiers, food additives, food components, and food substitutes as well as pharmaceuticals. Amino acid and peptide esters possess biological activities similar to those of lipoproteins found in nature, and are potentially useful as pharmaceuticals. The reactions above are depicted in the direction of reverse hydrolysis, which will dominate only under low water conditions, £.*. in hydrophobic, non-polar solvent. Since the hydrophilic compound is not miβcible with non- polar solvent, it is problematic to establish reaction conditions which are favorable to reverse hydrolysis. Moreover, most hydrophilic and hydrophobic compounds are immiscible in one another, and for many compound combinations there is no suitable solvent which would allow the mixing of both substrates. Consequently, esteri ications of many hydrophilic compounds, such as glycerol, ethylβnβglycol, polyols, and saccharides with long chain fatty acid residues, are difficult or impossible to achieve by traditional means in a batch or column reactor. For instance, the conven ional industrial process for the esterification of glycerol with fatty acids is conducted at 200-250'C in the presence of an inorganic catalyst, often a toxic metal. one class of amphiphilic products uβeful as e ulsifi β r β includes long-chain monoacylglycerols or monoglyce ides. Conventionally, they are produced by chemical alcoholysiε of the corresponding triglyceridβs with two equivalents of glycerol (see Figure 5B) . As in industrial •Ete ification, this type of reaction requires high temperatures (210-240 β c) and the use of tranβestβrification catalysts, usually toxic tin or lead compounds. The reaction product is an equilibrium mixture consisting of monoglyceride, glycerol as well as fatty acids and several contaminating by-products resulting from dehydration processes. In order to yield "pure" monoglycerides, as rβgioiβomerβ, expensive purification steps are required leading to product yields of only about 40-501 based on the starting glycerides. Alternatively, chemical ultistep processes are reported in the literature based on isopropylidene-glyceroi (Bear, E., Fischer, H.O. ., J. Am. Che . Soc. SI, 2031 (1945)) or glycidol (German Patent DE-OS 2.338.462 (1973)). In these methods, natural starting materials cannot be employed. All the above methods require the synthesis of suitable starting materials and/or protection/deproteetion steps. Thus, the above described conventional synthetic methods do not provide convenient means to obtain chemically or isomerically pure monoglycerides. A potential alternative 1B offered by biocatalyeiβ utilising enzymes (for an overview, see Wong, CH., science 244. 1145-1152 (1989)). In the chemical reactions discussed herein, both of the compounds changed in the reaction are considered substrates for the enzyme (Wong, supra_ . in order for enzymatic reverse hydrolysis to be made practical for industrial use, the above mentioned problems of substrate/solvent immisibility must be overcome. Catalysis of esterification reactions by the enzyme lipase has been described. It was reported that, in a reaction between glycerol and free fatty acids, mycelial Upases catalysed the ormation of ester bonds at positions 1 and 3 of glycerol to form mono- and diglycerides (Tahoun, M.K. j&Al., Fi-gym-- Mierob. Φeohnol. &, 429-432 (1966)). Reactions were carried out in aqueous medium, with dispersion promoted by stirring. Lipasβs have also been used in the esterification of saccharides with fatty acids (Bjδrkling, F. s£ al. f J.C.S. Chan. Conmun.. 934-935 (1989); AdelhOrst, . At Al., fiypthesis. 112-115 (1990)). The reactions were conducted without added solvent since the two substrates were somewhat soluble in one another. Water produced in the reactions was removed i-n vacuo. For the preparation of monoglycerides, enzymatic ultiβtep processes have been reported based on the starting compounds isopropylideneglycerol (Omar, I. ., Sacki, H., Nishio, N., Nagai, N., Blotechnol. Lett, ϋ, 161 (1989)) or glycidol (MUller, C, Austin, H., Posorskβ, ., Cruzalez, I., Novo Induatri Publication A-05991). These enzymatic methodβ have the same limitations as the chemical multi-step processes described above. In order to increase the rate of reaction and yield, lipase-catalyzed esterification in aqueous medium has been carried out in membrane bioreaotors. In the membrane bioreactor system, reverse hydrolysis is promoted through the use of microporous hydrophobic membranes which physically separate the glycerol/water/lipase solution from the fatty acid substrates (Yamanβ, T. t &1., Ann. y.Y. Aca . Bci. ϋl r 558-568 (1984); Hoq, M.M. fifc Al. . ϋ, 335-342 (1985)). The fatty acids penetrate the microporeβ of the hydrophobic membrane sufficiently to participate in the reaction at the interface of the membrane and the aqueous solution; the resulting amphiphilic products remain in the aqueous compartment. Another approach requires covalent linkage of substrate to a solid support in order to increase the exposure of substrate surface area to reactive groups. Enzymatic peptide synthesis has been promoted in aqueous medium by ester or amide bond linkage of amino acid substrate to activated silicagel (Koenneσke, A. fit l ' , Monatsh. Che . HS 331-337 (1982); Koennecke, λ. t Al-* Monatsh. Chem. 116. 111-117 (1985)). Lipase catalyzed reactions have also been carried out in reverse micellar systems or microemulsions. These systems were designed to promote the exchange of fatty acid side chains on triglycerides. The dispersion of triglycerides in polar solvent was achieved through e ulsification with surfactants. When the hydrophilic substrate glycerol was substituted for aqueous buffer in the emulsion system, no ester product was formed. (Kol berg, K. it £!• . 2QA££ ££, 1544-1548 (1968); Zakβ, λ., l&βfifi ϋ, 4β4 (1989)). Peptide synthesis was reportedly catalyzed by chymotrypsin in an organic solvent, hexane, in which neither the substrates nor the enyz e were soluble (KUhl, P. et fl.1.. Tetrahedron Lett.. H 5213-5215 (1990)). Product yields were inversely proportional to hexane concentration, and fell to zero at moderate concentrations of hexane. Although the authors could offer no conventional explanation for their results, they speculated that the undiβsolved substrates promoted the reaction through direct particle-particle contact. SHEET βycoβidβ transfer ie another type of reaction which may be βnzymatically catalyzed. Products of glycoside transfer include complex oligoβaccharideβ, glycoproteins , and glycolipids. Glycosyltransferases are not yet readily available, but the corresponding hydrolases, i-fi. glyσoβidases (Table 1, category 3.2.1) are commercially available. The glycosidasβ, 0-galactosidaβe, has been employed to catalyze reactions between saccharides and alcohols. The reported yields were low (25-35%) because the reactions were carried out in an aqueous medium in which the initially formed products were partly decomposed by the hydrol tic action of the enzyme. The reported procedures could not be carried out in non-aqueous medium because of __∑a__iscibility of substrates with non-polar solvents. The problem of enzyme insolubility in organic solvents has been addressed by the adsorption of peptidases and Upases to solid supports such as celite and silicagel (Ferjancic, A. fi A ., Aaial. Microbiel. Biotechnol. ££, 651-657 (1990),- Schuch, R. _&_£>, ftnnl. Microbtol. Biotechnol. 2£, 332-336 (1989) ; Eigtved, P. £& &λ- , Proc. World Conf. Biofcachnol. Fate and Oils. AOCS, 134-137 (1987); Ya ana a, S. __£__!., Methods in Ensvmol. 11$, 405-411 (1987)). Amounts of silicagel or celite sufficient to adsorb catalytic amounts of enzyme were either added to the reaction mixture, or, more commonly, were purchased with the e zyme prβ-adsorbβd onto the support. With stirring, the adsorbed enzyme was dispersed in the hydrophobic solvent. The hydrophilic substrate, however, remained ineffectively dispersed in the solvent which limited the rate of reaction and final product yield. The hitherto reported phoβphorylation of biologically important molecules like glycerol, dihydroxyaσetonβ, enolpyruvate, monosaccharides, and nucleosides was dependent on biological phospherylating agents, usually adencsine triphosphate (ATP) . These reactions reportedly are catalyzed by such enzymes as glycerol inase and hexokinase, end are carried out in aqueous medium, such processes are costly because ATP is expensive, is needed in molar amounts, and therefore must be recycled. Recycling of ATP involves a number of additional enzymatic steps and auxiliary reagents. In addition to limitations in yield and rate of reaction, the published methods leave unsolved the problem of producing isomerically and enantio-mβrieally pure products such aβ 1-monoglycerides. When glycerideβ are produced in the above described enzymatic methods, mixtures of mono¬ glycerides with di- and tri-glycerideβ are obtained. When monoglycerides are iβolated from the product mixture, they are always mixtures of regioiβo erβ and βtereoiso βrs. Monoglycerides are classfied according to their absolute configuration as follows. 1-sn-monoglyceridβ ■ (S) - 1 - monoglyceride 3-sn-ι.onoglycdridβ - (R) - 1 - monoglyceride 2-sn-raonoglycβride ■ achiral The "sn-" nomenclature is a classification which is exclusively used for glycerides. In the conventional production of monoglycerides as described above, generally mixtures of race ic 1-monoglycerides and achiral 2- glycerides are produced. Such mixtures could be termed as consisting of: (R.S) - 1 - monoglyceride + 2 - monoglyceride. Purification steps are required to yield chemically pure monoglycerides as mixtures of regioiso ers from which each regioisomer must be separated by tedious chromatogr&phic processes. These processes necessarily operate on a very small scale and are not practical for industrial production. SUMMARY Of THE INVENTION AND OBJECTS This invention relates to a method for producing an amphiphilic product through the enzyme catalyzed reaction of a hydrophilic substrate with a second substrate. According to the present invention, a hydrophilic substrate is dispersed in a second substrate with a selected enzyme so that the reactive groups of both substrates are contacted by the active site of the enzyme and an amphiphilic product is formed. The method comprises combining the hydrophilic substrate with sufficient solid support to substantially adsorb the hydrophilic substrate, and dispersing the Adsorbed substrate in the second substrate together with an enzyme which has an a finity towards both substrates and which catalyzes the formation of the product. If one of the substrates is a liquid, the reaction may proceed without added solvent. If neither substrate is a liquid, it is necessary to include a solvent in the method. It is therefore an object of this invention to provide methods to disperse a hydrophilic substrate in a non-polar or slightly polar solvent which is suitable for use in the processing of food, health care, and cosmetic products. An object of the invention is to provide methods for the production of pure regioiβomers of 1,3-diglycerides. A further object of the invention is to provide a method for the production of a sugar ester through the enzyme catalyzed reaction of a hydrophilic carbohydrate with a carboxylic acid derivative in a non-polar solvent. λ further object of the invention is to provide methods for the production and specific precipitation of selected 1-monogiycerides in chemically and isomerically pure forms. A further object of the invention is to provide methods for the production of esters of amino acidβ. A further object of the invention is to provide methods for the production of glycoβides through the enzymatically catalyzed transfer of glycoside moieties to a polyalcohol. A further object of the invention is to provide methods for the synthesis of peptides through the enzymatically catalyzed formation of peptide bonds. A further object of the invention is to provide methods for the enzymatically catalyzed synthesis of phosphates of alcohols, carbohydrates, and nucleosides. A further object of the invention is to provide compositions comprising alcohols or carbohydrates adsorbed onto a solid support such as silica gel. A further object of the invention is to provide compositions comprising first and second hydrophilic substrates, both adsorbed to a solid support. In such a composition, the first substrate is a carboxyl-protected amino acid or carboxyl-protected peptide with a free primary amino group, and the second substrate is an amino- protected amino acid or an amino-protected peptide with a carboxylie acid ester. BttTEF PBggRIPTION OF THE DRA INSS Figure 1 depicts the effects of different enzyme preparations on the rate of the esterification of glycerol with vinyllaurate. Figure 2 depicts the effects of different methods of adsorbing glycerol to solid support on the rate of esterification. Figure 3 depicts the effect of the order of mixing substrates, solid support, and enzyme on the rate of esterification of glycerol. SUBSTITUTE SHEET Figure 4 depicts the effects of dif erent methods of adsorbing βorbitol to solid support on the rate of esterification of sorbltol. Figure 5 schematically depicts the reactions which yield l-aonoglycerides. Figure € is a schematic illustration of a reactor/ separator system which may be used in the produotion of a pure l-aonoglyceride from a reaction which does not yield water or alcohol as a by-product and thus goes to completion without removal of products. Figure 7 is a schematic illustration of a reactor/ separator system which may be used in the production of a pure 1-monoglyceridβ from a reaction which yields water or alcohol as a by-product. The inclusion of molecular sieves to remove water or alcohol promotes the reaction in the desired direction of esterification. DETAILED INSCRIPTION OF THE PREFERRED EMBODIMENTS in the method of the invention, an enzymatically catalyzed reverse hydrolysis reaction is promoted between two substrates by adsorbing the hydrophilic substrate to a solid support and dispersing the adsorbed hydrophilic substrate in the second substrate with a selected enzyme. The hydrophilic substrate adheres to the solid support through a non-covalβnt interaction, possibly an electrostatic attraction, between the polarized hydroxyl group or amino group of the hydrophilic substrate and the polarized regions of the solid support. The second substrate contains a donatable leaving group (£.£. an acyl group) which probably combines transiently with the enzyme to form an acyl/enzyme intermediate (Wong, ___iE____) . It is postulated that, through mixing of the acyl/enzyme intermediate with the adsorbed hydrophilic substrate, the hydroxyl group or amino group of the hydrophilic substrate is brought in contact with the active site of the enzyme and with the donatable leaving group from the second substrate to form an amphiphilic compound. In an alternative embodiment related to peptide synthesis, both substrates contain a hydrophilic region, and both substrates are mixed together prior to adsorbtion onto the solid support. peftofaionff The term "hydrophilic substrate" refers to a water soluble compound containing a hydroxyl group or a primary amino group having the general structure: z can equal OH or N_ϊ 3 . x and Y can equal H, methyl, acyl, allyl, alkyl, aryl, or a inb groups. X and Y can also equal methyl, acyl, allyl, alkyl, and aryl groups which have an H substituted by a halogen or by sulphate, phosphate, nitro-, or oyano- groups. Examples of hydrophilic compounds are alkanols, cycloalkanolβ, benzyl alcohols, aHylic alcohols, diols, triols, polyols, sugar alcohols, inositols, halogen substituted alcohols, hydroxycarboxylic acids, amino acids, peptides, saccharides, synthetic and natural polymers with hydroxyl groups. The term "second substrate having a donatable leaving group" refers to compounds such as carboxylic acids, esters, anhydrides, amino acids with protected or unprotected side chains, peptides, proteins, phosphate esters, glycosides, and substantially hydrophobic alcohols. The donatable leaving group is a chemical moiety for which the enzyme to be employed has an affinity at the enzyme's active site, which can be detached from the second substrate, and which can be covalently linked to a hydroxyl or amino site on the first substrate. The donatable leaving group may be a chemical moiety such as an acyl group, an a ino acid residue, a peptidyl group, a protein residue, a phosphoryl group, a glycosidyl residue, or an alkoxide group. The term "amphiphilic product" refers to a compound which has both hydrophilic and hydrophobic groups. Examples of amphiphilic products include esters, sugar esters, peptides, peptide esters, glycosides, nucleosides, glycolipi≤β, glyooproteins, lipoproteins, phosphates, and sulphates. The term "ester" refers to the product of the reaction between a polyol, a carbohydrate, an amino acid, or a peptide and the donor of an acyl group such as a long chain fatty acid. The hydrophilic oities in the ester are polarized groups such as unreacted hydroxyl groups and the hydrophobic moities are, for instance, long carbon chains which have replaced a hydroxyl group on the starting substrate polyalcohol or carbohydrate. Examples of esters are mono-, di-, and triglycerides, which are the products of reactions between glycerol and donors of carbon chains such as free carboxylic acids, carboxylic esters and carboxylic vinylesters. The term "glycoside" refers to a compound which contains at least one sugar residue. Examples of glycosides are oligosacch& ides, glycoproteins, nucleosides, and glycolipids. The term "enzyme" refers to a proteinaceous catalyst capable of accelerating reverse hydrolysis reactions selected from the list in Table 1. Additional examples for enzymes useful in practicing this invention are found in the following references: Barman, T.E., Enzvme Handbook. Springβr-Verlag, N.Y. Vol II, pp. 602-S49, 1985; Sigma Chemical Company Catalog, St. Louis, HO, pp. 863- 869, 1991. The appropriate enzyme for a desired reaction product is chosen for its af inity and specific activity tovards both the hydrophilic substrate and the second substrate. The appropriate enzyme recognizes one or more of the donatable leaving groups of the second substrate according to the regioβpeci ic position and inherent characteristics of the leaving group, *.g. position 1, 2, or 3 of a triglyceride, chain length, degree of saturation. The appropriate enzyme also has an affinity for and reactivity towards the hydroxyl or amino group of the hydrophilic first substrate based on the size and local electrical charge of ohemical moieties surrounding the group. The enzyme may be obtained from a microbial source, or from plant or animal cells or tissues. The enzyme may be purified or included in a crude, relatively Impure mixture of several different types of enzymes. The enzyme preparation may be in any form such as a lyophilized powder, or covalently linked or adsorbed onto a solid support. Table .__: Enzymes which catalyze reverse hydrolytic reactions. Numbering and classification according to the convention of the International Union of Biochemistry (IUB) .Enzyme Nomftnclat-tra 197ft. Academic Press, New York 1979) . 2. TrftT.»f β rft6« 2.3 Acyltranβ erases 2.3.1 λcyItransferases 2.3.2 Aminoacyltransferases . Glycoβyltrans erases 2.4.1 HexosyItransferases 2.4.2 PentosyItransferases 2;7 Transferring phosphorus-containing groups 2.7.1 Phoβphotransferases with an alcohol group as acceptor 2.7.2 Phosphotransferases with a carboxyl group as acceptor Tablβ 1; (Continued) 2 * 7.3 phosphotranβ erases with a nitrogβneouβ group as acceptor 2.7.4 Phosphotranβferases with a phoβpho- group as acceptor 2.7.5 Phosphotransferases, apparently intramolecular 2.7.6 pyrophosphotrans erases 3. Hvdrolases 3.1 Acting on ester bonds 3.1.1 Carboxylic ester hydrolases 3.1.2 Thiolester hydrolases 3.1.3 Phosphoric monoester hydrolases 3 * 1.4 Phosphoric dlester hydrolases 3.1.5 Triphosphoric monoester hydrolases 3.1.6 Sulphuric ester hydrolases 3.2 Acting on glycosyl compounds 3.2.1 Glycoside hydrolases 3.2.2 Hydrolyβing N-glycosyl compounds 3.2.3 Hydrolyβing S-glycosyl compounds 3.3 Acting on ether bonds 3.3.1 Thioether hydrolases 3.4 Acting on peptide bonds (peptide hydrolases) 3.4.1 α-Amino-acyl-pβptide hydrolases 3.4.2 Peptidyl-a ino-acid hydrolases 3.4.3 Dipeptide hydrolases 3.4.4 Feptidyl-peptide hydrolases 3.5 Acting on C-N bonds other than peptide bonds 3.5.1 In linear amides 3.5.2 In cyclic amides SUBSTITUTESHEET Iftull_l: (Continued) 3.5.3 In linear amidines 3.5.4 In cyclic amidines 3.5.5 in cyanides 3.5.99 In other compounds 3.6 Acting on acid-anhydride bonds 3.6.1 In phoβphoryl-containing anhydrides 4.0 __X_L______ 4.1 Carbon-carbon lyaβes 4.1.1 carboxy-lyaβes 4.1.2 Aldehyde-lyaβes 4.1.3 Xetoacid-lyases 4.2 Carbon-oxygen lyases 4.2.1 Hydro-lyaβeβ 4.2.99 Other carbon-oxygen lyases 4.3 Carbon-nitrogen lyases 4.3.1 Ammonia-lyases 4 * 3.2 Amidine-lyases 4.4 Carbon-sulphur lyases 4.5 Carbon-halide lyases 4.99 other lyaβes The term "solid support" refers to a finely divided material having a large surface area compared to its volume, which is capable of adsorbing hydrophilic compounds. The support material is non-reactive with the substrate chemicals employed and is insoluble in the solvent, when a solvent is used. Examples of solid supports capable of adsorbing hydrophilic materials are silica gelsi diatomaoeous earths, clays, zeolites, activated charcoals, carboxymethyl cellulose and other substituted celluloses such as cellulose esters, neutral or basic ion exchange resins, porous glass beads, and neutral or basic aluminumoxide. λdeorption onto solid supports makes the hydrophilic substrates diβpersablβ through mixing in hydrophobic substrates. Alternatively, the solid support may be compressed into a stationary column onto which the hydrophilic substrate is adsorbed and through which the hydrophobic substrate is circulated. The solid support provides the basis for the prinicple of improving contact of the hydrophilic substrate with reactive groups within the icroenvironment of the enzyme/substrate interphase. The term "polar solvent" refers to a liquid compound in which one region of the molecule has higher electronegativity than a different region of the same molecule. Generally, hydrophilic compounds are soluble in polar solvents. Very polar solvents such as dimethylfor amidβ, dimethylβul oxide, dimethylacetamide, or pyridine are frequently unsuitable for use in esterification because they are toxic, difficult to remove and/or recycle. Residual concentrations would render the synthesized product unsuitable for food, cosmetic, and health care applications. Also, many enzymes are inactivated by polar solvents. The term "non-polar solvent" refers to a liquid compound in which the centers of positive and negative charge essentially coincide or cancel each other BO that no dipcle moment is produced, ∑n this application, the term "non-polar solvent" also includes those liquid compounds in which the centers of positive and negative charge substantially coincide so that only a small dipole moment is produced, £..&. slightly polar solvents. As a general rule, hydrophobic compounds are soluble in non-polar solvents, while hydrophilic compounds are not soluble in them. Examples of non-polar organic solvents are simple aliphatic hydrocarbons (n-hexane) , and aromatic hydrocarbons (toluene) . Examples of βlightly polar organic solvents are ethers, ketσneβ, and ββtβrs. Other examples of slightly polar solvents are chlorinated hydrocarbons ( ethylenechloride) , cyclic ethers (tetrahydrofuran) , βtβrically hindered alcohols (t- butylmethylβther) , and acetonitrile. Non-polar solvents offer advantages over very polar solvents in that the non- polar or βlightly polar solvents can be removed completely by conventional means; thus they are suitable for use in food, pharmaceutical, and cosmetic preparations. The above listed non-polar and slightly polar solvents do not inactivate Upases, as do some highly polar solvents. According to the method of the present invention, an ester can be prepared rapidly and conveniently by mixing the following: a hydrophilic substrate a solid support in an amount βufficient to adsorb the hydrophilic βubβtrate a second substrate having a donatable leaving group an enzyme catalyst a non-polar or βlightly polar solvent, if necessary or desirable for the specific reaction. Suitably, the first substrate is glycerol and the second substrate is a free carboxylic acid (fatty acid) which, upon reverse hydrolysis, will yield an amphiphilic ester product plus water. The second substrate may also be a methyl- or alkyl-ester which, upon reverse hydrolysis, will yield an amphiphilic ester product plus an alcohol. These types of reactions are termed "reversible", since the products could participate in the reverse reaction (hydrolysis of the desired ester product) if water or alcohol were allowed to accumulate. The invention may be utilized either with or without the removal of by- products. If water or alcohol is not removed from the product mixture, the reaction will go to equilibrium, which, through reverse hydrolysis or alcoholysis, will yield a certain amount of desired ester product. Preferably, however, water or alcohol is removed from the reaction mixture by the inclusion of a drying agent such as molecular sieves or anhydrous sodium sulphate, in order to allow the continuation of reverse hydrolysis. Alternatively, water or alcohol may be removed by azeotropic diβtillation in which the solvent mixture is empirically determined to yield selective distillation of water or alcohol. The method of the invention is also utilized in "irreversible" reactions in which no water or alcohol is produced, suitably, a second substrate such as a vinyl- carboxylic ester or an isopropβnyi-ester is reacted with a first substrate such as glycerol to form the desired ester product plus acetaldehyde or acetone. This type of reaction is termed "irreversible" because the acetaldehyde or acetone products cannot participate in a hydrolytic reaction. Thus the reaction proceeds in the desired direction of reverse hydrolysis without removal of the by-product. In both the above examples, the "donatable leaving group" on the second substrate is a chain of carbon atoms. According to the present invention, the second substrate having donatable leaving groups is mixed with the hydrophilic substrate adsorbed to solid support, in a low water medium, with the appropriate enzyme. The mixture is agitated at a temperature and for a time sufficient to produce a high yield of amphiphilic product. The amount of solid support used iε proportional to the amount of hydrophilic substrate. When the hydrophilic substrate is an alcohol or a carbohydrate, the ratio of solid support to substrate is preferably in the range of about 0.3:1.0 to about 10.0:1.0, more preferably 5.0:1.0, most preferably 1.0:1.0 (gsg). in the method related to peptide synthesis, both the first and second substrates are hydrophilic, and both substrates are adsorbed to the solid support, in this oaβe, the ratio of solid support to the combined substrates is preferably in the range of about 0.6:1.0 to about 10.0:1:0, more preferably 5.0:1.0, most preferably 1.0:1.0 (g:g). The ratio of solid support to hydrophilic substrate is selected to assure that at least about 10%, more preferably about 50%, most preferably about 100% of the hydrophilic substrate is adsorbed to solid support. The method of the invention is thus distinguished from previously published methods in which the amount of solid support used was only sufficient to adsorb the catalytic amount of enzyme used. For instance, Schuch et al ( su p ra ) reported adding only 5% (w/w) of immobilized enzyme preparation. Thus, even if it is assumed that the entire amount of the support upon which the enzyme was immobilized was available for adsorption of the hydrophilic compound, the proportion of support to hydrophilic compound was no more than 1:20 in the method of Schuch. In one embodiment of the invention, the hydrophilic substrate may be mechanically mixed with the solid support prior to mixing with the second substrate and the biocatalyst. In a second alternative, the hydrophilic substrate may be disolved in a minimum amount of a polar solvent such as methanol and mixed with the solid support. The hydrophilic substrate adsorbed to solid support is then isolated from the polar solvent by filtration or by evaporation, then mixed with the second substrate and the biocatalyst. In a third alternative, the hydrophilic substrate is mechanically mixed with the solid support and the enzyme to form a slurry, which is then packed into a column through which the second substrate is circulated. The method of the invention may be practiced with any order of mixing. Thus, the hydrophilic βubstrate may be pre-adsorbed to solid support as described above. Alternatively, the hydrophilic substrate, the second substrate, and the non-polar solvent, if necessary, may be pre-mixed to form an emulsion prior to addition of solid support and enzyme. The appropriate enzyme is chosen, according to the desired reaction, from the group listed in Table l. The enzyme preparation may be free in the form of lyophilized powder, or may be preadsorbed onto a different or the same type of support as the hydrophilic substrate, or may be covalently attached to support. Suitably, the enzyme is selected from among a family of Upases synthesized by microorganims such as Hueor mihel. Pseudomonas fluorescens. Pftjzopus deleaar. and £eni£i i___B cycloplum. A lipase is an enzyme which, in the presence of water, catalyzes the hydrolytic breakdown of fats. In a low water environment, certain lipases catalyze the reverse reaction, reverse hydrolysis, to yield ester products. Individual lipases have selective activity for regioisomeric and enantiomeric forms of the hydrophilic and the hydrophobic substrate. For instance, a 1,3- βpeci ic lipase from Mucor ihei can promote the production of esters formed exclusively at the 1 and 3 positions of glycerol. Alternatively, a less specific lipase, or a mixture of lipases, may be used in order to obtain triglycerides. Any lipase may be used provided the enzyme exhibits the required substrate speci icity or tolerance towards the position, the chain length, and the degree of saturation of the donatable acyl group. The selectivity of the enzyme varies from type to type and determines which types of products are formed. Certain lipases are capable of catalyzing the esterification of amino acids or peptides. The activities of specific lipases may be predicted from descriptions in the volume j_lpaBttB. «d. Borgβtrom, B. and Brock an, H. ., Elsevier, Amsterdam (1984). The enzymes known as glycosidases, j_.fl. 0-galactosidaβa, are capable of catalyzing the transfer of glycoεyl groups from glycosylated compounds to polyalcohols. The physical conditionβ for the reaction, such as temperature, stirring, and pressure, may be tailored aocording to desired rates and energy input. The temperature used is suitably between about -48*C and about 100 β C, and the reaction typically is run for between about 3 hours and about 3 days. The lower temperature limits are determined by the freezing point of the solvent or of the mixture. For instance, since the freezing point of hexane is about -50*C, the invention method using hexane would necessarily be conducted above -50 *C. The optimum temperature and time depend on the specific reaction and the enzyme selected, as illustrated by examples hereinafter. For industrial production, it may be desirable to run the reaction under less than optimal conditions because of savings in energy, or because it may be desirable to leave the reaction vessel unattended for a longer period of time. Thus, the method of the invention promotes convenient and rapid enzymatic reverse hydrolysis, producing high yields of commercially useful amphiphilic products. The progress of the reactions are measured by conventional means such as chro atography (thin layer, gas, high performance liquid, and the like). After complete or desired conversions of substrates to products have been achieved, the solid support and the enzyme are removed by conventional means such as simple filtration or centrifugation. Reaction products are isolated with high chemical purities by removal of the organic solvent through conventional means such as evaporation. * -22- In another embodiment of the invention a 1-mono-glyceride is selectively produced and precipitated. Suitably, a 1- onoglyceride is produced by the direct esterification of glycerol with a free carboxylic acid or with an alkyl or 5 vinyl ester (Figure 5A) . Alternatively, a 1-monoglyce id* may be produced by the esteri ication of glycerol with carboxylic acid residues from the alcoholysiβ of a triglyceride; the alcoholysiβ is catalyzed by the selected enzyme and occurs concomltantly with the esteri ication 10 reaction (Figure SB) . A suitable apparatus for performing this embodiment of the invention is illustrated is Figure 6. Central to this embodiment is the selection of a solvent or solvent mixture which, when cooled, promotes the selective 15 precipitation of the desired l-monoglyceride product. In this embodiment, the hydrophilic first substrate is glycerol, which may be adsorbed onto solid support. The glycerol is mixed in & reactor vessel 1 with a selected enzyme, the acyl donor, and the selected solvent. The 20 mixture is stirred in the reactor 1 at a temperature between about -_8 C C and about 100 e C. The temperature for the reactor is selected, according to the enzyme employed, to allow the enzymatic esteri ication activity to produce the desired l-monoglyceride and by-products. The mixture 25 is drawn through a filter disc 2 and transferred through a connector 3 by means of a pump 4 to a separator 5 which is cooled to a temperature which promotes the selective precipitation of the 1-monoglyceride. Under the employed conditions, all other products and unreacted substrates, 30 including 2-mono-, di- and triglycerides, are soluble and remain in the circulating solution. They are drawn through another filter disc 6 and transferred throught a retur connector 14 to the reactor 1 where they may again serve as acyldonors for the esterification of glycerol. 35 By selecting the optimum solvent mixture and the optimum temperature for selective precipitation, 1-monoglycerideβ which are chemically and isomerically pure can be obtained in very high yield without any further purification β tep. When the acyl donor is a vinyl or isopropenyl ester or a triglyceride, water or alcohol is not a product of the reaction and the reaction is irreverβible. n this case, the reaction will prooeed to completion in the desired direction of l-monoglyceride synthesis without removal of produots. When the acyl donor is a free carboxylic acid or a methyl- or alkyl-ester (Figure 5A; R' = H, methyl, alkyl) , water or an alcohol is a by-product of the reaction; the reaction is reversible and will go only to equilibrium if products are net removed. Figure 7 illustrates an apparatus in which water or alcohol by-products are removed in the practice of the invention. Molecular sieves 7 of the appropriate pore size are included in the connector 8 to remove water or alcohol and thus promote the reaction in the desired direction of esterification (reverse hydrolysis) . suitably, water is removed by molecular sieves of a pore size of 3 Λ, methanol by a pore size of 4 A. All other components in Figure 7 are equivalent to those in Figure 6, i.£. reactor 9, filter disc 10, pump 11, separator 12, filter disc 13, return connector 16. The molar ratios of glycerol and the corresponding acyldonor may be varied to produce variable mixtures of mono- and diglycerideβ in the reaction mixture. In order to maximize the yields of the deβired monoglyce ides, the stoichio etrically appropriate molar ratios of glycerol to acyldonors are employed (a»~S»- 1 ftole glycerol:l mole donatable acyl group) . here economically desirable, a molar excess of one substrate may be used. Any lipase may be used that is stable in the selected solvent and that is active towards the acyldonor employed. f a free fatty acid or its ester iβ the acyldonor, a lipase iβ selected that has specificity for the carbon chain characteristics of the acyl group, j_.fi- chain length, degree of saturation. For instance, lipases from A. niger and 6. candidum are essentially inactive towards lauric acid or laurate esters (see Example 3). If laurate is the acyl donor of choice, the lipase iβ suitably chosen from &____________ . R. delemar. or P, ftluprffgp? na., all of which are active towards laurate. If di- and triglycerides are to be employed as acyldonors, the regiospecificities of the lipases are considered. For instance, the lipase from fø_. mihei has specific activity for the acyl groups at the 1 and 3 positions of di- or triglycerides and leaves the acyl group at the 2 position unreacted. For efficient and quantitative consumption of the acyl donor, preferably lipases with activity also on the 2-position of glycerides are employed. Examples include lipases derived from yseudomonas species or from Penicilllum cvclopium. The optimum solvent mixture and optimum temperatures for reaction and precipitation are empirically determined for each desired ono-glyceride product. Suitably, when the desired product is a l-monoglyceride, the solvent iβ a mixture of t-BuOMe:n-hexane in the proportion of about 1:100 to about 1:10, prefereably 1:1 (vol:vol:). Alternatively, when a suitable acyldonor is employed, the solvent may be 100% n-hexane. The solvent composition is entirely dependent on the product desired. For production of l-monoglyceride, the temperature in the reaction vessel is between about -48*C to 100*C, more preferable 20*0 to 60"C, most preferably 25*c to 35*C. For specific precipitation of l-monoglyceride, the temperature in the separator is between about -49 β C to 15•C, more preferably between about -10*c to 10 β C, most preferably 2*0. SUBSTITUTESHEET λ further embodiment of the invention provides a method for the production of a peptide-ester through the lipase catalyzed reaction of a peptide or an amino acid with a vinyl-ester. Suitably, the alpha-amino groups of the amino acids are protected by the covalently bonded Z-group (Z = carbobenzyloxy) . The amino aoid or peptide is adsorbed to solid support and reacted with a donatable source of acyl group such as a vinyl-carboxylic acid. Through the catalytic action of the lipase, the carboxylic acid residue is covalently bound to the hydroxyl residue of the amino acid or peptide to form an amino-acid-ester or peptide ester. A further embodiment of the invention provides methods for the enzymatic production of glycosides such as oligosaccharldea, glycoproteins, and glycolipids. Suitably, a hydrophilic first substrate containing a glyccsyl moiety is adsorbed to solid support and mixed with a second βubstrate such as a polyalcohol, and a glycosidase, such as β-galactosidaβe. The enzyme catalyzes the transfer of glycosyl moities from the first substrate to the hydroxyl groups of the second substrate to form the desired glycoside product. A further embodiment of the invention provides methods for the enzynatic synthesis of peptides. The hydrophilic first substrate in these reactions is a carboxyl-protected amino acid or peptide with a free primary amino group. The term "carboxyl-protected n refers to an amino acid or peptide in which each carboxyl-group is covalently linked to a group such as a tert-a yl ester, a benzyl ester, or an alkyl ester, which prevents the carboxyl group from participating in the reaction. The term "free primary amino group 0 refers to an unsubβtituted amine such a R-NH-, where R refers to an organie residue bonded by a carbon-nitrogen bond to the amine. -36- In the peptide synthesis embodiment, the second substrate is an amino-protected amino aoid or amino-protected peptide with at least one carboxylic acid ester. The term "amino-protected" refers to an amino acid or a peptide in which each amino group is covalently linked to a protecting group such as an acetyl, an amide, a tert- butyloxycarbonyl, or a trifluoroacetyl group, which prevents the amino group from participating in the reaction. The first substrate and the second substrate are dissolved together in a polar solvent, mixed with a solid support such as silica gel, and both substrates adsorb to the solid support The polar solvent iβ then removed under vacuum. The combined firβt and second substrates and solid support are then mixed with a non- polar solvent such as n-hexane and a peptidase such as alpha-chymotrypsin. The enzyme catalyzes the aminolysls of the carboxylic acid ester to make a peptide bond to form the desired peptide product. A further embodiment of the invention provides methods or the synthesis of a glycerol phosphate. Suitably, a hydrophilic substrate such as glycerol is adsorbed to a solid support such as silica gel and mixed with a non- polar solvent such as t-butylmethylether. To this mixture iβ added a phosphate donor such as a phosphate ester of an alcohol or of a phenol, or a carboxylic acid-phosphoric acid mixed anhydride. The reaction is catalyzed by a phosphatase such as acid phosphatase from potato. The phosphatase catalyzes the transfer of the phσsphσryl moiety to the primary alcohol functions of glycerol to form the glycerol phosphate. This method may also be applied to the enzymatically catalyzed synthesis of phosphates of other alcohols, of sugars, and of nucleosides. Examples of phosphate group donors which may be employed are n-buty phosphate, ethoxycarbonylphoβphate, and vinylphosphate. Examples of other hydrophilic substrates which may be employed are dihydroxyacetone, glucose, and thymidine. The enzyme catalyst may be an alkaline phosphatase, for instance, from E. coll. such reactions may produce, for instance, dihydroxyacβtone-l-phosphate, D-glucoβe-6-phosphate, or thymidine-5'-phosphate, respectively. A further embodiment of the invention provides compositions of hydrophilic substrates adsorbed to a solid support. When the adsorbed substrate is an alcohol or a carbohydrate, the ratio of substrate to solid βupport iβ preferably in the range of about 1.0:0.3 to about 1.0:10.0, more preferably 1.0:5.0, most preferably 1.0:1.0 (g:g) . The composition may also comprise two substrates adsorbed to the solid support, as in a composition used for peptide synthesis. In this case, the ratio of the combined substrates to solid βupport is preferably in the range of about 1.0:0.6 to about 1.0:10.0, more preferably 1.0:5.0, most preferably 1.0:1.0 (g:g). In the compositions, the hydrophilic substrate is dispersed in a thin layer of molecules on the surface of a finely divided solid βupport such as silica gel to provide a large exposed surface area of substrate. In compositions having two substrates adsorbed to the solid support, the substrateβ are also dispersed in thin molecular layers on the surface of the βupport. it is postulated that this dispersion allows the adsorbed substrate(β) to undergo rapid two-dimensional diffusion from the hydrophilic support through lipophilic interfaces to reach the enzyme active site. Preferably, the surface of the solid βupport is fully saturated with the hydrophilic substrate(s) in order to present the maximum number of reactive molecules to the enzyme, thus speeding the reaction. On the macromolecular level, the new compositions, comprising one or two hydrophilic substrateβ adsorbed to the solid βupport, take the form of a free- flowing powder. SUBSTITUTE SHEET The actual ratio of substrate(s) to solid βupport in a given composition can be determined using methods well known in the art of organic chemistry. For instance, the composition can be heated at S00*c for several hours, and the loss of weight determined in comparison to a control (solid support without substrate) . The following examples illustrate specific embodiments of the present invention. EXAMPLE I This example demonstrates the ef icacy of various types of solid support. One gram of glycerol was mechanically mixed with 1 g of silicagel (230-400 mesh, Merck) , or other solid βupport (see Table 2) until a homogeneous, free flowing powder was obtained. The glycerol adsorbed to solid support was then mixed with 0.4 gm of vinyllaurate as acyldonor, and 100 g of immobilized preparation of lipase from Mueor ipjftf j (Lipozyme, Novo) , in 10 ml of t-butylmethylether (t-BuOMe) . As a control, the same amounts and types of substrates, biocatalyst, and solvent were mixed without added solid βupport. The mixtures were stirred for the indicated number of days (see Table 2) at 25'C at atmospheric pressure. The enzyme was removed by filtration and the products were recovered from the organic phase by evaporation of the organic solvent. The products were analyzed qualitatively by thin-layer chro otography (TLC) , followed by quantitative analysis using gas chromotography (GC) . The control mixtures (i.fi. without solid support to adsorb glycerol) remained biphaslc and the enzymes precipitated at the intβrphase leading to a gel which proved impossible to recover. Thus, no conversion was observed in the control mixtures without added support after 7 days. Results were expressed as the percent of vinyllaurate converted to mono- di- and triglycerides (Table 2) . The proαuct composition was a consequence of the regioselectivity of the biocatalyst, which in this case, the lipase from Mucor mlhei. was relatively selective for the one and three hydroxyl groups of glycerol, reβulting in a preponderance of mono- and diglyceride producta. As βhown in Table 2, the most efficient supports were silicagel (Aeroβil) , Floriβil, and activated charcoal. However, all the supports listed are potentially uβeful. Tablβ 2. (Continued) Enzymic esterifications of glycerol on different in order to determine the effects of different solvents on the rates of conversion of glycerol to products, the experimental protocol of Example 1 was repeated, using silicagel as solid support, and using the solvents listed in Table 2. The reactions were carried out at 25*C for 24 hours, after hich the percent molar conversion of vinyllaurate to productβ waβ meaβured as in Example 1. The activity was calculated as % ol&r conversion divided by time. Results are listed in Table 3 as relative activity, based on the reaction rate with n-hexane as 100%. SUBSTITUTE SHEET λβ shown in Table 3, a vide variety of solvents were suitable for the reaction. Due to their high relative activity, n-hexane, toluene or t-BuθMβ may be the solvents of choice for many types of reaetionβ. The other listed solvents may be useful in oases where the acyldonor iβ insoluble (or little soluble) in hexane, toluene, or t- BuOMe. .able 3. Esterifications in organic solvents EXAMPLE 3 These experiments were designed to determine which types of lipase preparations would be sufficient to catalyze an esteri ication reaction according to the method of the invention. EXAMPLE 3A_ Glycerol (0.92 g, 10 mmol) was mixed mechanically with 1.0 g Silicagel (Merck 230-4007 mesh) until the glycerol liquid was completely absorbed. The tfree flowing, dry powder was mixed with 10 g t-BuOMβ and 5.42 g (24 mol) vinyllaurate (molar exceββ) to which waβ added either 150 g of crude lipase from Paaudomonas fitioreseens (Amano Pharmaceutical Co., Kagoya, Japan) or 324 mg of Lipozyme. The reaction catalyzed by Lipozyme waβ nearly complete after 3 hours. Products from the Lipozyme reaction were analyzed by TLC. After removal of the solid components by filtration and during evaporation of the solvent 4.9 g (100%) of crude l.3-dilaurin crystallized out. Recrystallization from n-pentane produced 4.6 g (95%) of pure 1.3-dilaurin, white σryβtalβ mp 57°C. Percent yields were calculated based on molar conversion of glycerol to product. The reaction mixture from P. fluoreβcenβ was filtered after 30 hourβ, the filter was washed twice with 20 ml tBuOMe and the solvent was removed in vacuo (rotovapor) . The resulting crude product mixture (4.7 g, 96%) was separated by LC on Si0 3 producing 0.5 mmol (5%) Trilaurin, 8.54 mmol (85%) 1.3-ϋilaurin and . mmol (11%) onolaurin. Percent yields were calculated based on molar conversion of glycerol to products. EXAMPLE 3B: in order to determine the time dependence of eβterifica ion reactionβ on the βpecific enzymes employed, the following experiments were carried out using the same conditions aβ outlined in Example 1 (molar excess of glycerol) with the exception that the following enzyme preparations were used: RhJZPpUfl d-άST-flE, £a___4i____ eylindraeea. Panieillium evclooium (Amano Pharmaceutical Co., Nagoya, Japan). The mixtures wre βtirred at room temperature at 750 rpm and samples were drawn at the times Indicated in Figure 1 for analysis (TLC, EtjO:n-hexane, 1:1, I--development, verified by GC) . -34- The reaction catalyzed by lipase from p. delemar was essentially complete by 45 hours, as only traces of the original starting amount of vinyllaurate remained. The lipase preparations from P. cvclopiitm and c. cvllndracea 5 were somewhat less efficient as the reactions were about 80% to 90% complete after 100 hours. SXΛKP E 4 This experiment was designed to test whether the method of the invention could be scaled up for industrial use. 10 Glycerol (46 g, 0.5 mol) was mechanically adsorbed on 46 g silicagel and suspended in 1 liter of tBuOKe to which 226 g vinyllaurate had been added. After addition of l.o g Lypozyme, the mixture was stirred at room temperature for 48 hours. After removal of the solid components by 15 filtration and evaporation of the solvent, 226 g of crude glycerides (85% 1,3-dilaurin) were obtained. The giycerides were dissolved in n-hexane and the solution waβ filtered over 300 g silicagel. The reβulting solution was cooled to -30 β C resulting in 160 g (70%) of 1,3-dilaurin; 20 the remaining products were recovered from the mother liquids. By recryβtallization from n-hexane, 1,3-dilaurin was obtained in isomerically pure form (see Example 1 above) . 25 This example demonstrates the preparation of glycerides from various vinylesterβ. Glycerol (10 mmol) was adsorbed to l gm of silicagel, then suspended in 10 ml tBuOMe. Vinylester (vinylvalerate, caprylate, laurate, or palmitate) was added in the amount 30 of 20 mmol, after which 50 mg of Lipozyme was added. The mixtures were stirred for 24 hours at room temperature. After removal of the enzyme and silicagel, the solvent was evaporated to yield crude products. The crude productβ were dissolved in n-hexane, filtered over silicagel and cooled to -30 β c. The resulting mixture contained 85-90% of 1,3-diglycerides with >95% purity. EXftrø___E.6 This experiment waβ designed to test the effect of inclusion of molecular sieves to remove water from the reaction mixture during the direct esterification of glycerol with fatty acids. Carboxylic acids [caprylio acid, C8; lauric acid, C12; palmitic acid, C16; 20 mmol each] and 10 mmol of glycerol preadsorbed to 1 gm of silicagel were suspended in 20 ml of tBuOMe. After addition of 50 mg immobilized lipase from Mucor mihei and 3 g 3 A molecular sieves, the mixture waβ stirred for 48 hours at room temperature. After removal of all solids by filtration, the organic phase was washed with NaHCOj-solution to remove traces of the fatty acids. After the separation steps described in Example 3, 1,3-diglycerides were obtained typically in yields from 65-85%. BXWE 7 This experiment was designed to demonstrate the esterification of ethylene glycol. Ethyleneglycol (0.621 g, 10 mmol) was mixed mechanically with 1 g Silicagel (Merck 230-400 mesh) until the liquid was completely absorbed. The free flowing, dry powder was mixed with 12 g t-BuOMe and 4.75 g (21 mmol) vinyllaurate to which 150 mg of crude lipase from Faeudomonaa sp. was added. The mixture was stirred at room temperature at 750 rp for 8 hours after which the reaction was complete. TLC analysis showed no trace of starting material, .£. quantitative conversion of substrates waβ achieved. Ethylβneglycoldilaurate in the amount of 4.0 gm (91%) was isolated. . 36 _. EXAMPLE 8 This example demonstrates the production of polyesterβ. Ethyleneglycol (0.621 g, 10 mmol) adsorbed on 1 g of Si02 (silicagel) waβ reacted with 2.0 g (10.09 mmol) of divinyladipate in 10 g of t-BuOMe in the presence of 150 mg of crude lipase from PBeudomonas fluorttscens. After 8 days the mixture was analyzed and found to contain a mixture of polyesters. EXMffLE 9 This experiment waβ designed to assess the dependence of the reaction rates on the method of preparing the adsorption of hydrophilic substrate. Percent adsorption was determined by collecting a sample of the silicagel/glycerol complex and (a) extracting the glycerol from the silicagel by washing with water, and quantifying the amount of glycerol by conventional chromotographic or enzymated methods, or alternatively by (b) drying at room temperature under vacuum and determining the weight increase. Preparation 1.1. As described in the above examples, glycerol was mechanically mixed with an equal amount of silicagel (230-400 mesh, Merck) until a homogeneous, free flowing powder was obtained. Preparation 1.2. The glycerol waβ diββolved in a minimum quantity (lg/10 ml) methanol (HeOH) to which an equal amount of silica gel waβ added. After stirring for two hours the silica gel waβ isolated by filtration and the filter cake dried in air. only about 70% of the original starting amount of glycerol remained adsorbed on the silicagel in this preparation. Preparation 1.3. The glycerol waβ mixed with a minimum quantitiy of acetonitrile (lg/ Oml) and adsorbed onto silicagel. The βilioagel waβ isolated as in 1.2 above. nly about 65% of the original starting amount of glycerol remained adsorbed on the silicagel. Preparation 1.4. The glycerol was mixed with MeOH and adsorbed onto silicagel as in reparation 1.2 above. The KβOH waβ removed from the βuspension on a rotavapor in vacuum. Preparation 1.5. This preparation waβ the same as Preparation 1.4, with the exception that acetonitrile was used in place of MeOH. All glycerol preparations were then esterifled using 50 mg of an immobilized preparation of lipase from Mucor ihei (Lipozyme, Novo) as biocatalyst and 2.0 g vinyllaurate as acyldonor, in 8.0 ml t-BuOMe with 0.5 gm glycerol adsobed on silicagel. The reactions were monitored via the consumption of vinyllaurate (Figure 2). As shown in Figure 2, all the above preparations yielded satisfactory results. Preparation 1.4 (adsorption from MetOK) produced the most homogeneous βilicagel/glycerol preparation with the best handling characteristics. it should be noted that a hydrophobic solvent such as t- BuOMβ was not essential for the esterification in this example since the acyldonor, vinyllaurate, was a liquid. The adsorbed glycerol, together with the biocatalyst, may be dispersed directly in the liquid second substrate. The rate of conversion, however, may be different. EXAMPLE 10 This experiment was designed to assess whether it waβ necessary to pre-adsorb the hydrophilic βubstrate before conducting the reaction. Preparation 2.1. This was the control preparation in which the glycerol waβ pre-adβorbed on the βilicagel aβ in Example 9, Preparation 1.1. Preparation 2.2. In a 10 ul flaβk 0.47 g (5 mmol) of glycerol, 2 g (10 mmol) of vinyllaurate and 8 ml of t-BuOKe were thoroughly mixed. To the formed emulsion, 0.5 g of βilicagel were added and the mixture waβ stirred for about 1-2 minuteβ. Then 50 mg of Lipozyme waβ added and the resulting mixture waβ stirred for a total of 24 hourβ. At regular intervale, samples were drawn and analyzed. Preparation 2.3. As in Preparation 2.2, the glycerol, vinyllaurate, and t-BuOME were mixed to form an emulsion. In this case, the Lipozyme waβ added and stirred for 1-2 minutes prior to adding the βilicagel. The reβultβ (Figure 3) βhowed that rates of oo verβion were comparable among all three preparations. Preparation 2.1 waβ the fastest; however, when βilicagal was added later (2.2 and 2.3) the reaction rate was satisfactory due to adsorption of glycerol in situ onto the silicagel. This experiment was designed to determine whether solid, hydrophilic alcohol could be esterifled using the method of the invention. Preparation 3.1. Sorbitol was mechanically mixed with an equal amount of silicagel using a mortar and pestle until a free flowing, homogeneous appearing mixture waβ obtained. Preparation 3.2 Sorbitol waβ diββolved in a minimum quantity of 99% MeOH to which an equal amount of ailioagel was added. After stirring for 2 hours, the silicagel was recovered by filtration and dried, only 45% of the original starting amount of sorbitol remained on the βupport. Preparation 3.3. Sorbitol waβ dissolved in a minimum quantity of 99% MeOH to which an equal amount of βilicagel waβ added. The solvent waβ then removed under vacuum uβing a rotavapor. The residue, a free flowing powder, waβ used directly in the esterification reaction. Sorbitol (0.8 g, corrected for the amount remaining after adsorption), 0.4 g vinyllaurate, 50 mg immobilized lipase from Mucor mihei (Lipozyme, Novo) and 8 ml t-BuOMe were mixed and stirred for a total of 24 hours. At regular intervals samples were drawn from the mixture and analyzed by TLC (Figure 4). From all three adsorbed preparations, esterification was observed. Clearly the fastest reaction waβ observed with the material prepared according to method 3.3 (adsorption from MeOH and solvent removal under vacuum) . EXAMPU- 12 This experiment was designed to assess the effect on esterification of the ratio of solid support to glycerol * Glycerol (0.92 g, 10 mmol) waβ adβorbed on varying amounts (0, 0.3, 0.6 and 1 g) of βilicagel (Merck 230-400 mesh) and esterifled as in Example 1 in t-BuOMe with 5.42 g (24 mmol) of vinyllaurate as acyldonor. AB esterification catalyβt, 150 mg of lipaβe derived from Pβeudomonaβ fluoreβcenβ were employed. After an incubation time of 30 hours all reaction mixtures were analyzed by TLC and worked up for quantitative product isolation. Only the samples containing 0.6 and 1 g of βilicagel showed complete conversion of glycerol to ester product. The resultβ are summarized in Table 4. For efficient conversion, it iβ preferable to uβe a βilicagel:glycerol ratio greater than about 0.6:10 (g.mmol) . ______________* EXAMPLE P.3 This experiment was designed to aββess whether a solid carbohydrate, glucose, could be esterifled according to the method of the invention. D-gluoose (0.412 g, 2.29 mmol) adsorbed on 0.588 g of silicagel was added to a mixture of 20 ml t-BuOMe and 30 mmol of either vinylacetate, vinylbutyrate, or vinyllaurate as acyl donor. After addition of 200 mg lipase derived from Pseudomonas luorescens the mixtureβ were βtirred for 14 dayβ. The reaction mixtureβ were ther. iltered and the filter cakeβ were washed thoroughly with fresh t-BuOMe. The solvent waβ removed on a rotavapor and the reβidue dried on a high vacuum line. From the reactionβ using vinylacetate as acyl donor, 370 mg of a yellow syrup were obtained and separated by chromotography yielding three fractions: (1) 75 mg syrup RF-0.96 (front); d-glucoβe-2,4,6- triacetate. (2) 185 mg syrup RF-0.79; d-glucσβe-2,6-diacetate. (3) 100 mg colorleββ cryβtalβ, RF-0.54; D-glucoβe-6- acetate. (Solvent for chromatography: BtOλo: 2-propanol:^0-600:270:130; Nuclear Magnetic Resonance [NKR] analysis conducted at 400 MHz.) From the reaction using vinylbutyrate aβ acyldonor, 580 mg of a colorless syrup was obtained. Separation by chromatography on βilicagel yielded 60 mg crystalline material, RF=0.69; d-glucose-6-butyrate. From the reaction using vinyllaurate as acyl donor, 5.8 g of a colorless liquid remained, conβisting mainly of excess vinyllaurate and lauric acid. TLC analysiβ revealed that mainly product(β) of low polarity with RF-0.94 had been formed. After a few dayβ of βtanding at room temperature, the mixture crystallized and the cryβtalβ were isolated by filtration. NM analyaiβ demonstrated the formation of D-glucoβe-6-laurate. Aβ a control, esterification of D-glucose without solid support was attempted. 1.8 g (10 mmol) of anhydrous D-glucoβe waβ added to a mixture of 4.6 ml (50 mmol) vinylacetate in 20 ml of t-BuOMe. After addition of 300 mg lipase derived from Pβeudomonaβ fluoreβcenβ the mixture waβ βtirred at room temperature for 14 dayβ. After that the reaction mixture waβ filtered and the filter cake waβhed with fresh t-BuOMe. The solvent waβ removed on a rotavapor and the residue dried on a high vacuum line, obtained were 60 mg of a colorleββ syrup, corresponding to a yield of lees than 5% based on conversion of D-glueose. In summary, the resultβ from thiβ experiment indioated that D-glucose oould be eβterified according to the method of the invention. The main productβ were D-glucoββ-6- acetate, o-gluooβe-2, 6-diacetate, and D-glucoβe- ,4,6- triacetate. The yieldβ ranged from 30% to 50%. EXAMPLE 14 This example demonstrates the production of isomerically and chemically pure 1-monoglyceride by direct esterification of glycerol with free carboxylic acid. In this context, the term "chemically pure" refers to a preparation of l-monoglyceride substantially free of 2-monoglycerides, diglycerideβ, and triglycerides. The term "isomerically pure" refers to a preparation of 1- onoglyceride substantially free of 2-monoglycerides. The amounts and types of substrates and silicagel used are indicated in Table 5. The glycerol was preadsorbed onto the silicagel as in Example 3, then mixed in the reactor vessel (Figure 7,7) with the indicated carboxylic acid and 100 mg of Lipozyme βuβpended in 80 ml of a 1:1 mixture of t-BuOMe/n-hexane. The reactor 7 waβ connected with the separator 11 via a column illed with 3 A molecular sieves 6 to remove water produced in this reaction. The reaction mixture was stirred in the reactor 7 at a temperature of 25*C at atmospheric pressure. The separator (Figure 7,11) was cooled to 2*C. The solution was circulated continuously from the reactor 7 (maintained at 25*C) to the separator 11 (maintained at 2*C), and back to the reactor 7 at a rate of 1 a / in. Two filter diβcβ, one in the outlet from the reactor 8 and one in the outlet from the separator 12, were employed to retain solid materials, λ precipitate waa obβerved forming in the cooled separator 11. At the times indicated in Table 5, the reaotion waβ terminated and the product precipitated in the separator 11 waa collected by simple suction filtration. The preolpitate was washed with two volumes of cold n-hexane and analyzed by gas chromotography of the trimethylsilylether (TMS) derivative. The TMS derivative waa produced by reacting a hydroxylgroup with trimethylβilylchlorlde to produce the volatile TMS derivative for analyβiβ by gas chromatography. The pure l-monoglyceride productβ were obtained with iβomerio and chemical purities greater than 95%. Aβ βhown in Table 5, column 1, when glycerol waβ used in excesβ, 90% yield waβ obtained within 48 hours (percent yield baβed on molar converβion of lauric acid) . When βtoichiometric amountβ of reactanta were used (columns 2 and 3), the reactionβ progreββed more slowly; a yield of 86% waβ obtained by 120 hours (Table 5) . TABLE 5 1 » lauric acid β * βtearie acid EXMP E S Thiβ experiment demonβtrateβ the production of chemically pure -monolaurin by eβterifioation of glycerol with vinyllaurate. The amountβ of substrates and silicagel are indicated in Table 6. As in Example 3, the glycerol was preadsorbed to the silicagel, then mixed in the reactor (Figure 6,1) with the vinyllaurate and loo mg of Lipozyme in 80 ml of a 1:1 mixture of t-BU0Mβ/n-hexane. The reactor 1 was connected with the βeparator 5 aβ βhown in Figure 6. Thiβ system differβ from that uβed in Example 14 only in the elimination of the molecular sieves in the connector. Molecular sieveβ are not neceββary in this reaction, since water or alcohol iβ not a by-product. The βolution waβ βtirred in the reactor l at a temperature of 25*c at atmospheric preββure. As in Example 14, the separator 5 waβ cooled to 2*C and the βolution waa circulated through the system at a rate of 1 ml/min for the indicated time. The precipitate waβ collected, washed, and analyzed as in Example 14. Aβ shown in Table 6, the pure 1-monolaurin product waβ obtained in yields of 27% to 90% with iβomeric purity greater than 97% aβ determined by gas chromatography analysiβ of the TMS derivativeβ. TABLE 6 EXAMPLE 16 This experiment waβ designed to test the method of the invention in the larger scale production of l-monolaurin through the enzymatic esterification of glycerol with methyllaurate. Glycerol (23 g, 250 mmol) waβ adsorbed on 25 g of βilicagel as in Example 1. Mβthyllaurate (53 g, 0.25 mol) and 500 mg Lipozyme were suspended with the adsorbed glycerol in 500 ml of a 1:1 mixture of t-BuOMe/n-hexane in the reactor depicted in Figure 6. Molecular sieves of pore βize 4 A were employed to remove the methanol by¬ product, thereby promoting the reaction in the desired direction of reverβe hydrolysis. Aβ in Example 14, the βeparator waβ cooled to 2*C end the βolution waβ circulated through the system. The precipitate waβ collected from the βeparator and analyzed aβ in Example 14. After 48 hours, 17.8 g (26% yield) 1-monolaurin waβ collected froa the separator. After 144 hourβ, 45.9 g (67% yield) of thiβ material was produced with >99% iβo eric and chemical purity. EXAMPLE X7 Thiβ example demonβtrateβ the production of pure 1-monolaurin by esterification of glycerol with the triglyoeride trilaurin aβ acyldonor. The experiment waa conducted aβ in Example 15, with the exception that inβtead of Lipozyme, 100 mg of immobilized lipaβe from P. fluoreaoana was used (Amano Pharmaceutical Co, Ltd., Nagoya, Japan). Lipaβe from P. fluoreβeana has activity towards all three positions of the triglyoeride and thus effectively removes all the aoyl groups from the acyldonor. In contraβt, the lipase from M. mihei (Lipozyme) exhibits βpeoificity towardβ the 1 and 3 poβitionβ of the triglyoeride and would not be able to e fectively remove the .aoyl group from the 2 poβition of trilaurin. The amountβ of βubβtrateβ and βolid βupport are indicated in Table 7. As βhown in Table 7, yieldβ of between 21% and 80% were obtained with an iβomeric purity of l-aonolaurin greater than 95%. When glycerol was used in molar excess (column 1, Table 7) , the reaction proceeded rapidly to 80% yield within 48 hours. When stoichiometric amounts of reactantβ were employed (column 2, Table 7), the reaction proceeded more slowly to 75% yield after 96 hourβ. SUB TIT T TABLE 7 EXAMPLE 18 This example demonstrates the production of pure l-monopalmitin by the esterification of glycerol with methylpaImitate. The experiment was conducted as in Example 15 using 5.4 g (20 mmol) of methylp Imitate, 1.85 g (20 mmol) of glycerol adsorbed on 1.9 g of βilicagel and 100 mg of Lipozyme in 100 ml of the solvent mixture used in Example IS. After 48 hours, 2.31 g (35% yield) of l-monopalmitin with >97% isomeric purity waa obtained. EXAMPLE 19 This example demonstrates the uβe of solid βupport in a column as an alternative to the methods of preparation of adsorbed substrate employed in Example 9. Glycerol (3.7 g, 20 mmol) waβ mechanically mixed with 4 g of βilicagel and 100 mg of Lipozyme. The mixture waβ packed into a glaββ column (8 mm x 15 cm). λ βolution of 9.0 g vinyllaurate in 20 ml of t-BuOMe waβ circulated via a membrane pump through the column at a rate of 5 ml/mln for a total of 24 hσura. The column waβ then flushed with 20 ml freβh t-BuOMe and all organic phases were removed. The remaining viβcouβ material waa dissolved in n-hexane and flaβh chromatographed through a βhort silica gel colu n. The βolution waβ cooled to -20*C, which cauβed crude 1,3 dilaurate (about 95% pure) to crystallize out. After recryβtallization from n-hexane, 8.2 g (90% yield) 1,3-dilaurin was obtained as white crystals with a chemical purity of greater than 99%, aa determined by gaβ chromatography. EXAMPLE 20 This example demonstrateβ the production of a peptide- ester. The hydrophilic βubβtrate employed waβ Z-L-ββrinβ (Sigma Chemical Co., St. Louis, MO). The "Z N moiety iβ the carbobenzyloxy protection group which iβ covalently linked to the alpha-amino group of βerine in order to prevent the formation of zwitterions and in order to provide only one nucleophilic group for acceptance of the acyl group. -L-βerine (0.4785 g, 2 mmol) waβ adsorbed on 2 g ailicagβl uβing acetone aβ βolvent aβ deβcribed in Example 9, preparation 1.4. The reβulting free-flowing powder was mixed with 20 mol t-BuOMe and 10 mmol of vinylvalerate (1.28 g) or 10 mmol of vinyllaurate (2.26 g) . To thiβ mixture waβ added l g of Lipozyme. The mixture waβ shaken at 350 rpm at 30*C for 24 hours, after which all βolidβ were removed by filtration. From the reβulting organic phase, the βolvent waβ removed by evaporation (rotavapor) and the resulting residue was flash chromatographed on silicagel to remove excesβ vinylvalerate or vinyllaurate (n-hexane:t-Bu Me, 1:1, followed by acetone ethanol, 4:1). After removal of all βolvent the βolid reβidue waβ recryβtallized from n-hexane. Obtained from the reaction with vinylvalerate: 0.576 g (94 % yield) of Z-L-βerine valerate as colorless crystals, m.p. 188-189*0; l«] 2 .? - -5.0" (c« l. CHCl . Obtained from the reaction with vinyllaurate: 0.776 g (92% yield) of z-L-βerine laurate as colorless powder, .p. 160-162 » C; EXAMPLE 21 Thiβ example demonstrates the production of a glycoside, l-0-/J-D-galatopyranosyl-n-dodecanol. The hydrophilic substrate, o-nitrophenylgalacto-pyranoside (10.2 g, 32 mmol) iβ dissolved under ultraβonication in 150 ml of methanol warmed to 50°-60*C. The dissolved substrate iβ adsorbed onto 30 g of silica gel aβ described in Example 9, preparation 1.4. After removal of the βolvent, the material iβ dried under vacuum to produce a free-flowing powder. The material iβ then mixed with 320 ml of n-hexane and 17.6 g (96 mmol) of n-dodeσanol. The enzyme 0-galaotoβidaβe (2750 U, Sigma Chemical Co., St. Louie, MO) is added and the mixture iβ βtirred at room temperature for 24 hourβ. The βolid materialβ are removed by filtration. From the organic phaβe, 11.0 g (60 mmol) of n-dodecanol and 4.4 g (31 mmol) of o-nitrophenol are recovered. The solid material iβ thoroughly extracted with 3 x 200 ml of methanol and all βolid materials are removed by filtration. After removal of the βolvent, 9.46 g (26 mmol) of 1-0-0-D-galactopyranøay -n-dodecanol is obtained as semi-solid oily material. EXAMPLE 22 Thia example demonstrates the synthesiβ of N-acetyl-L- tyroβine-L-phen lalaninamide. N-Acetyl-L-tyrosine ethyleβter monohydrate (lmmol; 269 mg] and L-phenylalaninamide hydrochloride [lmmol; 201 mg] are diββolved in 20 al MeOH. To thiβ βolution iβ added 5 g of silica gel, and the MeOH ia removed under vacuum. The _ _ thua obtained material iβ mixed with 0 ml n-hexane, N jCOj.10 HjO [a mmol; 572 g], and 20 mg alpha- chy otrypβin. The mixture iβ βtirred for 24 hourβ at room temperature, and the βolvent iβ then removed by vacuum. The reβidue iβ then extracted with hot acetonitrile. HPLC analyβiβ of the reβulting βolution indicateβ a converβion of greater than 90% at thiβ point. The βolution iβ concentrated by removing the remaining βolvent under vacuum and the residue iβ recryβtallized from acetonitrile. The reβulting product iβ N-acetyl-L- tyroaine-L-phenylalaninamide in the amount of 259 mg (70%). EXAMPLE 23 Thiβ example demonstrates the enzymatically catalyzed βyntheβiβ of N-acetyl-L-tyroβinβ-L-leucinamide. N-λcetyl-L-tyroβine-ethyl eβter monohydrate (1 mmol; 269 mg) and L-leucinamide hydrochloride (l mol; 167 mg) are βuβpβnded in 20 ml t-Bu Ke. To thiβ βuβpenβlon, Na 3 CC_.10 HjO (2 mmol; 572 mg) , βilicagel (5 g), and alpha- cnymotrypβin (20 mg) are added. The reβulting mixture iβ βtirred for 24 hours at room temperature. After removal of the βolvent, the reβidue iβ extracted with hot acetonitrile. HPLC analyβiβ indioateβ a oonverβion of ■ greater than 95%. The iβolated dipeptide (80%) iβ identified by comparison with a known sample. gXWLE 34 Thiβ example de onβtrateβ the enzymatically catalyzed βyntheβiβ of glycerol-l-phoβphate. Glycerol (10 mmol; 0.92 g) iβ adsorbed to an approximately equal amount of βilica gel (1.0 g) and then βuβpended in 20 ml of t-BuOKe. To thiβ mixture iβ added p- nitrophenylphoβphate (10 mmol) followed by aoid phosphatase from potato (1000 U; 200 mg). The mixture iβ βtirred at room temperature for 24 hourβ. The βolvent iβ UB TITUTE re oved by vacuum, the reβiduβ iβ thoroughly extracted with water (3 X 150 ml) , and all βolidβ are removed by filtration. The resulting agueouβ βolution iβ concentrated under vacuum to 75 ml and treated with a βolution 21 mmol of BaCl 2 .2HjO to form a precipitate. To the βuβpenβion iβ added 500 ml of 95% ethanol and the precipitate iβ allowed to βettle overnight. The βuβpenβion iβ filtered and the βolid material iβ dried over CaS0 4 . Glycβrol-1-phoβphate (8.5 mmol) iβ obtained containing 94% of the title compound aβ the barium βalt.

Claims: WHΛT fg Cfaft-JHBD IS> 1. λ method for producing an amphiphilic compound comprising mixing a hydrophilic firβt βubβtrate containing at leaβt one hydroxyl group, a βolid βupport capable of 5 adβorbing said firβt βubβtrate, a second βubβtrate having at leaβt one donatable leaving group, and an enzyme, the ratio of βaid βolid βupport to βaid firβt substrate being about 0.3:1.0 to about 10.0:1.0 (g:g), βaid firβt βubβtrate adhering to βaid βolid βupport, βaid enzyme 10 catalyzing a reaction between βaid hydroxyl group and βaid leaving group to form an amphiphilic compound. 2. The method of Claim l further comprising adding an organic βolvent βeleoted from the group conβiβting of n-hexane, toluene, t-butylmethylether, ethylene oxide, 15 ethyllβobutylketone, vinylacetate, ethylacetate, t-butanol, acetone, diohloromethane, tetrahydrofuran, and acetonitrile. 3. The method of Claim 1 wherein βaid amphiphilic compound iβ an eβter and βaid firβt βubβtrate iβ βeleoted 20 from the claββ of compounds conβiβting of alcohols* and carbohydrates. 4. The method of claim 1 wherein βaid βecond βubβtrate is βelected from the claββ of compounds oonβiβting of carboxylic acids, esters, anhydrides, and triglycerides. 25 5. The method of Claim 1 wherein βaid enzyme iβ a reverse hydrolaβe. 6. The method of Claim 5 wherein βaid reverse hydrolaβe iβ a carboxylic eβter hydrolaβe. 7. The method of Claim 3 wherein βaid firβt βubβtrate iβ 30 βelected from the group conβiβting of glycerol, ethylene glycol, βorbitol, and glucoβe. 8. The method of Claim 4 wherein βaid βeoond βubβtrate iβ βelected from the group conβiβting of vinyllaurate, vlnyladlpate, and vinylacetate. 9. The method of Claim 4 wherein βaid βecond βubβtrate iβ βelected from the group conβiβting of caprylic acid, lauric acid, and palmitic acid. 10. The method of Claim 1 further compriβing the βtep of mechanically mixing βaid irβt βubβtrate with βaid βolid βupport before adding βaid βecond βubβtrate and βaid enzyme. 11. The method of Claim 1 further compriβing the steps of dissolving βaid irβt βubβtrate in a polar βolvent to form a βolution, mixing βaid βolution with βaid βolid βupport, and isolating βaid βolid βupport with βaid firβt βubβtrate adβorbed thereon before adding βaid βecond βubβtrate and βaid enzyme. 12. The method of claim 11 wherein βaid polar βolvent is βelected from the group oonβiβting of water, methanol, acetonitrile, and acetone. 13. λ method for producing an amphiphilic compound 'compriβing mixing a hydrophilic firβt βubβtrate containing at leaβt one hydroxyl group, a βolid βupport capable of adβorbing βaid firβt βubβtrate, and an enzyme to form a mixture, packing βaid mixture into a column, and circulating a βecond βubβtrate having at leaβt one donatable leaving group through βaid column, the ratio of βaid βolid βupport to βaid firβt βubβtrate being about 0.3:1.0 to about 10.0:1.0 (g:g) , βaid firβt βubβtrate adhering to βaid βolid βupport, βaid enzyme catalyzing a reaction between βaid hydroxyl group and βaid leaving group to form an amphiphilic compound which iβ eubstantially free of βaid βolid βupport. 14. The method of Claim 1 wherein βaid amphiphilic compound iβ a phosphate, βaid firβt βubβtratβ iβ βelβcted from the group conβiβting of alcohols, carbohydrates, and nucleosides, and aaid enzyme iβ a phosphatase. 15. The method of Claim 14 wherein βaid firβt βubβtrate iβ glycerol. 16. The method of claim 14 wherein βaid βecond βubβtrate iβ βelected from the group conβiβting of phosphate esters of alcohols, phosphate esters of phenols, and carboxylic acid-phoβphoric acid mixed anhydrides. 17. The method of Claim 16 wherein βaid amphiphilic compound iβ glycerol-l-phoβphate and βaid βecond βubβtrate iβ βelected from the group conβiβting of p-nitrophenylphoβphate, n-butlyphoβphate, vinylphoβphatβ, and methylcarbonylphoβphate. 18. The method of Claim l further comprising incubating βaid reaction mixture under sufficient physical conditionβ and for a time βufficient to achieve at leaβt about 10% converβion of βaid leaving groupa to βaid amphiphilic compound. 19. The method of Claim 1 wherein βaid βolid βupport iβ seleoted from the group conβiβting of βilica gelβ, diatomaceouβ earths, clays, activated charcoals, carboxy ethycβlluloβe, cellulose eβterβ, ion exchange resins, and insoluble polysaccharides. 20. λ method for producing a peptide compriβing, mixing a hydrophilic firβt βubβtrate having a free primary amino group, a βecond βubβtrate having at leaβt one donatable leaving group compriβing a carboxylic acid eβter, a βolid βupport capable of adβorbing βaid firβt and βaid βecond substrates, and a peptidaβe, the ratio of βaid βolid βupport to βaid firβt and βecond substrates being about 0.6:1.0 to about 10.0:1.0 (g:g) , βaid firβt and βaid βecond substrates adhering to βaid βolid βupport, βaid peptidaβe catalyzing a reaction between βaid free primary amino group and βaid carboxylic acid eβter to form a peptide. 21. The method of Claim 20 wherein βaid firβt substrate iβ βelected from the class of compounds oonβiβting of carboxyl-protected amino aoldβ and carboxyl-protected pβptideβ. 22. The method of Claim 18 wherein βaid βecond substrate iβ βelected from the class of compounds conβiβting of amino-protected amino acids and amino-protected peptides. 23. The method of Claim 20 wherein aaid peptidaβe iβ alpha-chymotrypβin. 24. The method of Claim 20 wherein βaid firβt βubβtrate iβ L-phenylalaninamide or L-leuβinamide and βaid βecond βubβtrate iβ H-acetyl-L-tyroβine ethylester. 25. λ method for producing and selectively precipitating an isomerically and chemically pure l-monoglyceride compriβing a) mixing glycerol βubβtratβ, a βecond βubβtrate, and a irβt organic βolvent to form a firβt mixture, βaid βecond βubβtrate having at leaβt one donatable leaving group, and βaid firβt organic βolvent being capable of promoting selective precipitation of a specified 1- monoglyσβride; b) combining βaid firβt mixture with an enzyme to form a βecond mixture, βaid enzyme catalyzing a reaction between a hydroxyl group of βaid glyoerol and βaid leaving group to form a l-monoglyceride; c) incubating βaid βecond mixture at a temperature βufficient to yield diββolved l-monoglyceride product; end d) cooling the mixture of step (c) to a temperature at which βaid 1-monoglyceride selectively precipitates. 26. The method of Claim 25 wherein after step (d) at leaβt βome by-productβ remain in a βolution, βaid by- productβ being capable of βerving aβ third βubβtrates by donating one or more acyl groups in βaid reaction and further compriβing the steps of β) recycling βaid βolution to a reactor veβββl, βaid reactor veββel having a βufficient amount of glycerol, βaid enzyme, and a temperature βufficient to allow for continuation of. βaid reaction to yield a third reaction mixture compriβing l-monoglyceride and by- productβ/ f) continuing βaid reaction; g) repeating steps (d), (e), and (f) until at least about 80% conversion of acyl groups to l-monoglyceride and precipitation of l-monoglyceride are obtained. 27. The method of Claim 26 wherein one by-product of βaid reaction iβ water or an alcohol, βaid water or alcohol being removed from βaid βecond mixture by βanβ of a drying agent. 28. The method of Claim 26 further compriβing adding to βaid firβt mixture a βolid βupport capable of adβorbing βaid glycerol, the ratio of βaid βolid βupport to βaid glycerol being about 0.3:1.0 to about 10.0:1.0 (g:g), βaid glyoerol adhering to βaid βolid βupport. 29. The method of Claim 27 wherein βaid βecond βubβtrate iβ a free carboxylic aoid. 30. The method of Claim 29 wherein βaid free carboxylic acid iβ lauric acid or atearic acid. 31. The method of Claim 25 wherein βaid βecond βubβtrate iβ a methyl eβter. 32. The method of Claim 31 wherein βaid methyl eβter iβ βthyllaurate or methylp Imitate. 33. The method of Claim 25 wherein βaid βecond βubβtrate iβ a triglyceride. 34. The method of Claim 33 wherein βaid triglyoeride iβ trilaurin. 35. The method of Claim 25 wherein βaid firat mixture iβ maintained at a temperature from about 48*C to about lOO'C. 36. The method of Claim 25 wherein βaid temperature of step (d) iβ from about -49*C to about 10°C. 37. The method of Claim 36 wherein βaid temperature is 2»C. 38. The method of Claim 25 wherein βaid βolvent iβ a mixture of t-butylmethylether;n-hexane in the proportion of about 1:100 to about 100:1 (volivol). 39. The method of Claim 38 wherein βaid proportion iβ 1:1 (vol.vol) . 40. The method of Claim 1 wherein said firβt substrate iβ βelected from the claββ of compoundβ conβiβting of amino acidβ, peptideβ, and proteinβ. 41. The method of Claim 40 wherein βaid amphiphilic compound Iβ an amino acid eβter and βaid firβt βubβtratβ 1B an amino acid. 42. The method of Claim 41 wherein βaid amino acid iβ Z- L-eβrine. 43. The method of Claim 42 wherein βaid βecond βubβtrate iβ vinylvalerate or vinyllaurate. 44. The method of Claim 1 wherein βaid amphiphilic compound iβ a member of the group conβisting of oligoβaccharideβ, glyooproteinβ, and glycolipidβ. 45. The method of Claim 44 wherein βaid firβt βubβtrate iβ a βaccharide and βaid enzyme lβ a glycoβidaβe. 46. The method of Claim 45 wherein βaid βaccharide iβ lactoβa. 47. The method of Claim 45 wherein βaid glycoβidaβe iβ 0-galactoβidaβe. 48. The method of Claim 44 wherein βaid βecond βubβtrate iβ a polyalcohol. 49. The method of Claim 48 wherein βaid polyalcohol iβ iβopropylidβnβglycβrol. 50. λ compoβition compriβing an alcohol or a carbohydrate adβorbed to a βolid βupport in the ratio of about 1.0:0.3 to about 1.0:10.0 (g.g) . 51. The compoβition of Claim 50 wherein βaid alcohol ia a polyol, 52. The oompoβition of Claim 51 wherein βaid polyol Iβ βelected from the group conβiβting of glycerolβ, glycolβ, erythrotol, and βorbitol. 53. The composition of Claim 50 wherein βaid carbohydrate la gluooβe or galactoβe. 54. The compoβition of Claim 50 wherein βaid solid βupport iβ βelected from the group conβiβting of βilioa gelβ, diato aceouβ earths, clays, activated charcoals, earboxymethycβllulose, cellulose esters, ion exchange reβinβ, and insoluble polyβaccharideβ. 55. The compoβition of Claim 50 wherein said ratio is 1:1 (g:g). 56. A composition compriβing glycerol adβorbed to βilica gel in the ratio of about 1.0:0.3 to about 1.0:10.0 (g:g). 57. Die composition of Claim 56 wherein said ratio is 1:1 58. λ compoβition compriβing a hydrophilic firβt βubβtrate adsorbed to a aolid support and a second βubβtrate adsorbed to βaid βupport, βaid firβt substrate having a free primary amino group and being selected from the class of compounds conβiβting of carboxyl-protected amino acids and carboxyl-protected peptides, and said second substrate having at leaβt one carboxylic acid ester and being βelected from the class of compounds consisting of amino-protected amino acids and amino-protected peptides, the ratio of said solid βupport to said first and βaid βecond substrates being about 0.6:1.0 to about 10.0:1.0 (g:g). 59. The composition of Claim 58 wherein said solid support is βelected from the group consisting of βilica gβlβ, diatomaceouβ earths, clays, activated charcoals, σarboxy ethycelluloβe, cellulose eβterβ, ion exchange reβinβ, and insoluble polyβaccharideβ.

Inventors: BERGER MATTHIAS, LAUMEN KURT E, SCHNEIDER MANFRED P, BERGER, MATTHIAS, LAUMEN, KURT, E., SCHNEIDER, MANFRED, P.
Applicants: BERGER MATTHIAS, LAUMEN KURT E, SCHNEIDER MANFRED P, BERGER, MATTHIAS, LAUMEN, KURT, E., SCHNEIDER, MANFRED, P.
Application year: 1992
Main classification: C12P1
IPC codes: C12P1, C12P7, C12P9, C12P13, C12P19, C12P21