USE OF TYPE II ENDOGLYCOSIDASES IN BAKING FOR IMPROVING THE QUALITY OF DOUGH AND BAKED GOODS

Abstract: A method of improving dough characteristics is provided which indludes the steps of mixing dough with a Type II endoglycosidase to form a dough mixture, shaping the dough mixture, and baking the said shaped dough mixture to form a baked good. The method of the present invention preferably uses endoglycosidase H mixed with bread dough which imparts better make-up performance of the dough, superior crumb qualities, softness of bread and higher loaf volumes.

Description: USE OF TYPE II ENDOGLYCOSIDASES IN BAKING FOR TMPPΩVJNG THE CUALTΥ OF DOUGH AND BAKED GOODS Field of the Invention The present invention relates to the use of endoglycosidase baking for jjrtproving the characteristics of baked goods. More particularly, the present invention involves the use of Type II endoglycosidases in baking to impart superiorcrumb qualities, andto inprovemakeupperformance in dough, softness of bread, and volume. Background of the Invention The "Straight Dough" and the "Pre-Ferment" methods are two major conventional methods for the maniifacture of yeast raised bread products. For the straight Dough method, all of the flour, -water or other liquid, and other ingredients-whichmayinclude, butarenotli tedtoyeast, grains, salt, shortening, sugar, yeast nutrients, dough conditioners, andpreservatives are blended to form a dough and are mixed to partial or full development. The resulting dough is allowed to ferment for a period of time ranging f om 0 to 6 hours or more depending upon specific process or desired end-product characteristics. For example, "no-time" processing may call for virtually no fermentation time, or just enough time to convey the dough mass to the next processing stage. "Short-time" processing may require a somewhat longer fermentation (i.e., 10-60 nύnutes) . "Long-time" processing may require fermentation times over 60 minutes to achieve desired results. The next step in the process is the mechanical or manual -division of the dough into appropriate size pieces of sufficient weight to ensure achieving the targeted net weight after baking, cooling, and slicing. Some systems, particularly commonwith roll production, call fora continuous degassing step and possibly final developing step .immediately prior to dividing. The dough pieces are often then rounded and allowed to rest (Intermediate Proof) for varying lengths of time. This allows the dough to "relax" prior to sheeting and molding preparations. The time generally ranges from 5-15 minutes, but may vary considerably depending on specific process.ing and formulations. The dough pieces are thenmechanically ormanually formed into an appropriate shape. This is often accomplished by passing the dough through one or more sets of sheeting rolls to flatten the dough piece, followed by a curling operation to form an elongated dough cylinder. Numerous variations of the molding operation exist. In same cases, doughs are processedthrough "string" lines in which long dough cylinders are then cut into portions which,later become roll products. For pan bread, the molded dough pieces are transferred into greased pans. In the case of hearth bread, the dough pieces may be transferred onto screens, pans, sheets, or other appliances for further processing. The doughpieces are then given a fnal "proof" prior to baking. Thetimes varywidelydependingonprocess anddesired fnishedproduct. Times may range from zero to several hours. Typical ranges for most wholesale pan bread and rolls is 40 to 60 minutes, 90-100°F, and 85-95% relative humidity. The dough pieces are then baked at various times, temperatures, and steam conditions in order to achieve the desired endproduct. Loaves are then often cooled for up to anhour ormore prior to slicing (if desired) and packaging. Inthe PrB-Fermentmethod, yeast is combinedwithotheringredients andallowed to ferment for varying lengths of time prior to final mixing of the bread or roll dough. Baker's terms for these systems include "Water Brew", "Liquid Ferment", "liquid Sponge", and "Sponge/Dough". Apercentage of flour ranging from 0-100% is combined with the other ingredients which may include but are notlimitedtowater, yeast, yeastnutrients anddough conditionersandallowed to fermentundercontrolledorambientconditions foraperiodoftime. Typical times range from 1-5 hours. The ferment may then be used as is, or chilled and stored in bulktanks or troughs for lateruse. The remaining .ingredients areadded (flour, characterizingingredients, additional additives, additional water, etc.) and the dough is mixed to partial or full development. The dαutgh is then allowed to ferment forvarying lengths of time. Typically, as some fermentation has taken place prior to the addition of the remaining ingredients, the time required is minimal (i.e., 10-20 minutes), however, variations are seen depending -upon equipment andproduct type. Follow.ing the second fermentation step, the dough is then treated as in the Straight Dough Method. As used herein the term "dough" or "doughmixture" describes a pliable mixture that minimally comprises a flour ormeal and a liquid, such as milk orwater. As used herein the term "dough ingredient" may include, but is not exclusive of, any of the follow.ing ingredients: flour, water or other liquid, grain, yeast, sponge, salt, shortening, sugar, yeast nutrients, dough conditioners and preservatives. Asusedherein, theterm "bakedgood" includes but is not exclusive ofall bread types, including yeast-leavened and chemically-leavened and white andvariety breads and rolls, english muffins, cakes and cookies, confectionery coatings, crackers, doughnuts and other sweet pastry goods, pie and pizza crusts, pretzels, pita and other flat breads, tortillas, pasta products, and refrigerated and frozen dough products. The use of enzymes to improve dough characteristics in baking is well known in the art. For example, amylases are used to hydrolyze theβ -1,4-glycosidic linkages in polysaccharides, such as in starch granules. Alpha-amylases can attack long starch chains at random at their interior and can dextrinize and liquefy starch. The starch chains canthen be furtherbrokendownbyβ -amylase intomaltoseandmaltotriose. Theuseofamylasesprovidesmaltose forenhanced yeast fermentation and .induces changes in dough characteristics, such as a decrease in water absorption capacity, a slackening of the dough, and the development of stickiness. Alpha-amylases also exert an .indirect effect on crust color by releasing sugar during the early stages of baking which causes crust browning. Alpha-amylase alsobrings aboutstarchdextrinizationdur.ingtheearlystagesofbaking, which results in an improved grain and a softer texture. .Alpha-amylases are endoglycosidases which are reactivewith specific internal linkages within an oligo- or polysaccharide substrate. Such endoglycosidases are referred to herein as Type I endoglycosidases. Additional Type I endoglycosidasesthathavealsobeenused inbakingarecellulase, glucoamylase and pentosanase (also known as hemicellulase or arabinoxylanase) . Proteases have also been employed inbakingto impart doughmellowing effects. Fungal proteases canbe added to the sponge to allowthe enzyme to act on flour proteinsduringthefermentationperiodthatpermits areductionindoughmixing time, improvedmachinability, fasterproofingduetobettergas retention, and in loaveswithbetter symmetry and improvedgrain andtexture characteristics. Bakincr Science and Technolociv. Vol. 1 , Pyler, E.J., Siebel Publisiing Co., Chicago, 111., pp. 132-182 (1988) . Other enzymes used in the baking industry include oxidoreductases such as lipoxygenasewhichcatalyzesthecoupledoxidationofcarotenepigmentsofflour (therebybleachingthem) andofunsatxirated fattyacidsbyatmospheric oxygen. Theenzymeisusedextensivelybybakersto improvethewhiteness ofcrumb color of bread. Pyler, et al. supra at p. 165; Sumner, J.B., and Sumner, R.J., j. Biol. Chem. 134. 531 (1940). As the demand for "natural foods" increases, the baking industry is searching formethods perceived as "natural" to obtain the softness, texture, whiteness and shelf-life characteristics of bread previously achieved by the use of additives such as emulsifiers, and other additives such as potassium bromate which is used as an oxidant for bread. Potassium bromate is used extensively as an improver for bakery flour. Breadbakers add it directly as a powder or tablet, or theymay use it indirectly as an ingredient inyeast food and dough conditioners. Most of thebreadproduced in NorthAmerica todayhas potassium bromate added in one form or another. Potassium bromate has several disadvantages in that when combined with combustible materials such as wheat flour, oil and grease it forms a highly flammable mixture that can ignite and explodeifconfned. Inaddition, ingestionofhighlevels ofpotassiumbromate havebeen foundto cause acute stomach cramps. Food Chemi«cal News, p.10, June 1, 1989. A "clean label" (i.e., a label free from ingredients viewed as "harmful chemicals" such as potassiumbromate) is therefore highly desirable since the baking industry .is searching for alternatives to the chemical additives they rely on currently. Enzymes are the natural additives that bakers will most likely turn to. The use of enzymes that will impart better machinability characteri.stics to dough, suchas bettermake-upperformance (i.e., betterdividing, roundingand molding characteristics) , andwill result inbreadwith fner andwhiter crumb quality and softer texture, will be useful in the baking industry. Type II endoglycosidases, as used herein, are a category of endoglycosidases which are capable of cleaving specific internal glycosidic linkages found in glycoproteins. These endoglycosidases cleave all orpart of the carbohydrate moietyframaglycoproteindependingonthelocationof thereactiveglycosidic linkage * Lntheglycoprotein. ExamplesofTypeIIendoglycosidases includeendo- β -N-a∞tylglu∞saminidases (Endo-D, Endo-H, Endo-L, Endo-CI, Endo-CII, Endo-F-Gal type and Endo-F) , endo-α-N-acetylgalactosaminidase, endo-β -N- galactosidases, peptide-N-(N-acetyl-β •• ■ •glucαsamiπyl) aspergineamidaseF (PNGaseF EC3* 5*> !• 52) andglyσopeptideN-glycosidase (PeptideN-glycosidaseEC3» ^ 2** 18) . See, e.g., Tarentino, A.L., et al. Biochem 24, 4665-4671 (1985) ; Arakawa, M. , et al., J. Biochem., 76, 307-317 (1974) ; Plummer, T.H. et al., J. Biochem.259, 10700-10704 (1984); Tarentino, A.L. et al., Biochem. and Biophys. Res. Cαmm. 67, 455-462 (1975) ; Trimble, R.B. et al. Anal. Biochem. 141. 515-522 (1984) ; Tarentino, A.L. , et al.. Methods in Enzymolocrv, 138, 770-778 (1987) ; Plummer, T.H., et al., J. Biol. Che .. 256. 10243-10246 (1981); and "Glycoprotein and Proteoglycan Techniques" by J.G. Beeley, Chap. 6, pp. 153-300, Elsevier, Amsterdam, New York, Oxford (1985) . In addition to having a specificity for theinternalglycosidiclinkagesofglycoproteins, atleastoneendoglycosidase (endo-β -N-a∞tylglucosaminidase H, i.e., Endo-H) has also demonstrated a specificity which produces the cleavage of lipid-liriked oligosaccharides (Chalifour, R.J. etal. Archives ofBiochem. and Biophys.229, 386-394 (1983)) and reportedly di-N-acet^lchitobiose linkages in oligosaccharides and glycoproteins (Tarention, A.L. et al. J. Biol. Chem.. 249. 811-817 (1974). Such Type II endoglycosidases, in general, have been used primarily for analyticalpurposes, e.g., thedeterminationofproteinorcarbohydratesequence and/orthe structureand functionof specificglycoproteins. See, e.g., HSieh, P. et al. J. Biol. Chem.. 258. 2555-2561 (1983); and Geyar, R. et al., Eur. J. Biochem.. 143. 531-539 (1984). TypeII endoglycosidases, however, havenotbeenusedinbakingto .ύπprcfvedough and baked goods. Ihe references discussed above are provided solely for their disclosure prior to the filing date of the .instant case, and nothing herein is to be construed as an admission that such references are prior art or that the inventors are not entitled to antedate such disclosure by virtue of prior invention or priority based on earlier filed applications. Summary of the Invention It is an object herein to provide methods ' utilizing Type II endoglycosidases alone or in combination with other enzymes to impixive doughmachinability and baked goods characteristics such as finer crumb quality, whiter bread crumb quality, softness of the baked good, and higher loaf volume. It is a further object herein to provide formulations containing Type II endoglycosidases useful in practicing such methods. In accordance with the objects of the invention, methods are provided for improving dough characteristics comprising the steps of mixing a Type II endoglycosidasewithdoughingredientsto formadoughmixture, andbakingsaid dough mixture. TheTypeHendoglycosidasesusedhereincanincludeendoglycosidaseH (Endo-H) , orotherendo-β-N-acetylglucOsaιιιinidasessuchasEndo-F, Endo-L, Endo-C, Endo- CH, Endo-F-Gal type, Endo-F, and PNGase. The Type II endoglycosidase of the present invention can also include endo-«-N-a∞tylgalactosaminidase and the endo-β-N-galactosidases. Detailed Description of the Invention I the present invention, a method of improving bread dough characteristics is provided which lends better machinability characteristics to bread dough, allowing forbettermake-up performance, andresulting inbreadwith finer and whitercrumbquality. Themethodofthepresentinventionalsoimpartssoftness to the bread and higher loaf volume. In accordance with the objects of the invention, methods are provided for improvingdoughcharacteristics especiallybreaddoughcharacteristics. Since every bread making process is different, there is no one particular standard or best method for making bread and conditions vary according to ingredients and desired qualities. However a typical method with workable ranges which will .improve the bread dough upon incorporation of a Type II endoglycosidase comprises the steps of mixing bread formula ingredients to full development with a Type II endoglycosidase to form a dough, allowing the dough to rest for about 0 to 60 minutes at about 4°C to about 40 e C and about 50% to about 90% relativehumidity, preferably for about 40 minutes at about 30°C and about 75% relative humidity; scaling the dough into individual pieces of about 400-600 grams each, preferablyabout 430 grams each; proofingthedoughpieces forabout 0 to about 20 minutes, preferably for about 10 minutes prior to molding, at about 20°C and 50% relative humidity, molding and panning the dough pieces, proofing the άaj . for about 0 minutes to about 90 minutes at about 20°C to about 40°C and about 70% to about 90% relative humidity, preferably for about 40 minutes to about 45 minutes at about 36°C, and about 92% relative humidity; and baking the dough at about 150°C to about 350°C for about 7 to 30 minutes, preferably at about 218°C for approximately 18 minutes, to form bread. The method for improving bread quality involves the use of a Type II endoglycosidase which is capable of cleav.ing all or part of a carbohydrate moiety from a glycoprotein. Apreferred amount ofthe Type II endoglycosidase is from about 10 ppm to about 1,000 ppm. A preferred Type II endoglycosidase is Endo-H in a concentration of between about 25 ppm and about 100 ppm. Other TypeII endoglycosidases canbeusedwhich canbe selected fromthegroup consisting of Endo-F, Endo-L, Endo-CI, Endo-CII, Endo-F-Gal type, and PNGase. Additional Type II endoglycosidases can be selected from the group consisting of endo-β-N-acetyl galactosaminidase and endo-β -N-galactosidases. Themethod of the present invention can further include mixing bread dough with other enzyme preparations such as cellulase preparations containing arabinoxylanase .in addition to the Type II endoglycosidase. The Endo-H enzyme (EC 3.2.1.96) used in the examples herein was obtained commerciallyor fromE. coli orB. subtilis hoststransformedto expressEndo-H from Streptσmvces plicatus. Measurement of dough development and viscosity over time was performed by use of a farinograph. A farinograph is a physical dough-testing apparatus, used extensively by bakers and millers since the 1930's. The farinograph gives an indication of the relative mixing and absorption properties of the flour -and any additives to be used in the bread formula. Tracings made by a farinograph are called a farinogram. The followingExamples are illustrativeofthemethodof thepresent invention. Example 1 Endo-Hconcentrationin∞nceritratedbrothwas40g/liter. Enzymeconcentration was determined fcy using a standard enzyme preparation and assayed by using β -galactosi-dase labeled N-glycopeptide, (Boehringer Mannheim Bioch«3mica, Indianapolis, IN) . The standard was developed from enzyme with specific activity of 25 Uhits/mg. One unit is the amount of enzyme requiredto release 1 micrαmole of (3H)-dansyl-Asn-(Glc Nac) 4 -(Man) 6 at pH 5.5 at 37"C in 1 min (Tarentino et al. , supra) . The constant doutghwei«ght method (AACC 54-21) was used. In this method, dough weight is kept constant at 480 grams. Hardredwinter (HEW) patentwheat flour wasused. Theamounts of flourandwaterwereadjustedaccordingly. Alldoughs utilized 297.5 grams of flour and enough water plus enzyme to bring the total weightto 480.0 grams. This .createda doughofapproximately 61.35% absorption (not corrected to 14% moisture in flour) . Farinograph studies of Endo-H at 500 ppm and a control were done. Results of the data from the farinograms are presented in Table 1 below. DT = departure time MTI = mixing tolerance index Stab. •*** • stability Endo-H = * endoglycosidase H PT = peak time The main objective of the farinogram curve is to achieve optimization on the 500 Bϋ (Brabender unit) line (i.e., the industry standard correlated to best bake-out performance) . A curve that peaks below the line is σver-hydrated, a curve over the line is under-hydrated. In this study, the control curvewas optimized on the 500 line and absorption (%water inthe dough) was not changed for anyofthe enzymetreatments toallow the curve to reflect any changes in water hydration properties caused by the treatments. Departure time is the time required for the curve to leave the 500 line. The longer the DT, the stronger the flour. However, a decrease inviscosity during the running of the farinograph may result in a short DT as was seen with the Endo-H treatment. Stability of flour is indicated by several parameters: DT, MTI, stability and the 20 minute drop line. These values give an indication of the tolerance of the flour to the stresses of mixing. Less tolerant flours will have shorter DTs and stabilities, higher MIT values, and greater 20 minute drop lines. Another cause of "weakness" in farinogram stability values would be a drop in viscosily. Stability parameters were lower in the 500 ppm Endo-H treatment indicating a decrease in the viscosity of the dough. Peak time (PT) is indicative of dough development time; how long it takes for the gluten to reach optimum -elasticity. PT for the Endo-H treatment was increased, possibly indicating a strengthening effect on flour proteins. The 20 minute drop line indicates the reduction in dough viscosity with time as seen in the Endo-H treatment. Inthis example, theuse ofEndo-H reducedtheviscosity and increasedthe time required formaximum elasticity to develop. Based on this apparent mellowing effect on the dough, it was theorized that extensibility and thus sheeting characteristics of the doughs would be improved fcy the addition of Endo-H. Example 2 Aseries ofwhitebreadtest doughswere run todeterminethe effects ofEndo-H. For these tests, a verybasic liquid ferment white bread dough was used. The only other ingredients were flour, corn syrup, salt, shortening, dairy blend, yeast food, and oxidative additives (i.e., potassium bromate and iodate) . 1. Control — ■ Above Formula 2. 100 ppm Endo-H * plus above formula 3. 500 ppm Endo-H * plus above formula * Dough water was reduced in Tests 2 and 3 to compensate for liquid added by the enzyme preparation. The test doughs as set forth above were mixed to full development, about 13 minutes. Floor time (i.e., rest time) was 40 minutes at 30°C and 75% relative humidity. Doughs were scaled into 430 grampieces. Intermediate proofingwas for 10 minutes at ambient conditions priortomolding. Doughpieces were then sheeted, molded, and panned into 32 square inch pans. Proofing was done to a constantheight (3/4" abovepanlip) at about 36 β C and 92% relativehumidity. The doughs were thenbaked for 18 -minutes at 218°C. The results are set forth in Table 2. TABLE 2 Height in Λ Dough Feel vessel After Dough Feel Proof Grain Loaf Softness ** me Score Vol. C24 hrs.) It was noted that at 500 ppm, Endo-H produced a dough which rose less in the fermentation vessel, and had a very shiny, wet surface. Proof was normal indicating good fermentation. Loaf volume was higher than the control, and grain was nearly perfect. The texture, or silkiness of the grain was similar to what one would expect in a sponge/dough for this base foπnula with no emulsifiers or conditioners. At 100 ppmEndo-H, rise in the fermentationvesselwas normal, andmake-upwas good. Loafvolumewas higher thanthe control, but less than the 500 ppmtest. The grain was nearly as good as the 500 ppm test. Based onvisual observation of testversus control loaves, it was demonstrated that as the Endo-H concentration increased, loaf volume and crumb quality improvied. Thus, use of Endo-H irπproved dough handling properties, such as extensibility and make-up performance, and allowed for improvements in loaf volume and final product quality. Example 3 A bread test dough is run to determine the effects of Endo-F. The formulas, baking conditions and procedures used in this example are identical to those in Example 2 except for the replacement of Endo-H with the enzyme Endo-F and a change in enzyme concentration level. Test Dcucths 1. Control = Base and Liquid Ferment Formulas same as in Example 2. 2. Test — Above formula plus 50 ppm Endo-F. The feel of the Test dough is slightly more extensible than the Control and proofs to the same heitght as the Control, but in slightly less time. * Loaf volume for the Test bread is higher than the control and the grain and crumb quality of the Test bread is better in cσmpari≤on to the Control. Example 4 The effects of ENGaseF on dough andbreadquality are studied in this example. The formulas, baking conditions andprocedures usedherein again are the same as those disclosed in Example 2 except for the replacement of Endo-H with the enzyme INGaseF and a change in enzyme concentration. Test Doughs 1. Control = Base and Liquid Ferment Formulas same as in Examples 2 and 3. 2. Test = 50 ppm PNGaseF plus above formula. The Test dough feels slightly softer and JLs more extensible than the control andtheTestdoughproofsto equalheight oftheControl in slightlyless time. The Test bread loaf volume is higher than the Control and the grain and crumb quality of the Test bread is better than the control in that the grain is smoother than the Control and the crumb is softer than the Control. Example 5 A series of white bread doughs, havingthe base formula set forth below, were tested qualitatively using a short time straight dough process to de-bermine the effects of various doses of Endo-H to dough and bread quality. The dose range used was 0-1000 ppm Endo-H. One kilogram of flour was used per dough. Each dough yielded three loaves of bread. The other ingredients were water, salt, sugar, yeast, shortening, and a commercial bread improverpreparation (Serrol BF28 from Serrol Ingredients, Australia) containing eraulsifiers, oxidants and enzymes. This commercial preparationdoesnot containtype II endoglycosidaseenzymes. Thebase formula and Endo-H concentrations were as follows: Base formula grams flour 1000.0 water 620.0 salt 20.0 sugar 10.0 yeast 12.5 shortening 10.0 bread improver 12.5 Test Doughs 1. base formula (control) 2. base formula + 25 ppm Endo-H 3. base formula + 50 ppm Endo-H 4. base formula + 100 ppm Endo-H 5. base formula + 500 ppm Endo-H 6. base formula + 1000 ppm Endo-H - 40 g formula water, to compensate for the softening effect of the Endo-H at this dose. All ingredients were added to a high speed mixer (energy input set at 50mH) andmixed for 30 seconds on "slow" speed, 2.5 minutes on "fast" speed. As shown inTest 6, less liquidwasutilizedatthis dosagetocompensate forthegreater softening effect of the Endo-Hat thehigher concentrations. Doughs were given 5 minutes rest out of the mixer at about 34 β C. Scale weight was 480g. After molding, the dough pieces were given an intermediate proof of 10 minutes at 34.5°C, molded again, panned, and set in the proofer for approximately 50-60 minutes, about 38°C and 80-90% relative humidity. Loaves were proofed to constant height. The loaves were baked at 240 β C for 20 minutes. The visual and tactile characteristics of the resultant doughs and loaves of breadwerecomparedqualitatively. ThequalitativetestingindicatedthatEndo- H imparted favorable characteristics to white bread at all dose levels. The maincharacteristicwasthe softeningeffectondoughs. This allowedthedoughs to mold-up better, which in turn produced a bread with finer crumb qualities which felt softer to the touch and appeared whiter to the eye. There was a dose response in the series of test doughs with the bread and dough in that he 25 ppm to 100 ppm Endo-H containing doughs gave the most favorable characteristics. The dough in Test 3 at 50 ppm Endo-Hwas determined to give the best results in that it produced a soft, pliable dough with no tackiness detected at the molder. Example 6 A series of tests was run in a short-time straight dough white bread formula. The purpose was to test the effects of combining a Type II endoglycosidase enzymewithacommercial cellulasepreparation, suchas Cytolase 300, ondough andbread quality. The cellulase preparation used in this example, cytolase 300, was obtainedfrcmGenencorInternational, SouthSanErancisco, California. The base formula and procedure used were as follows: *Added as separate solutions (solution water is included in the absorption indicated above) . Absorption: same for all tests (i.e., 62% water) Mix Time: 16 minutes Floor Time: 20 minutes Scaling Wt.: 430g for 32 in. pan Bench Time: 10 minutes Proof to 3/4" above pan lip Bake 425 β F for 18 minutes Test Doughs 1. Control: Base formula 2. Base formula + 100 ppm Endo-H 3. Base formula + 50 ppm cellulase preparation 4. Base formula + 100 ppm Endo-H + 50 ppm cellulase preparation 5. Base formula + 50 ppm Endo-H + 50 ppm cellulase preparation om ensate l test doughs containing enzymesweremore extensible thanthe control. Test Doughs 4 and 5, containingthe-mixtureof enzymes, required slightly lessproof time and had a slightly softer crumb 24 hours after baking than the other doughs. Loaf volume was also highest in test dough # 5. The best results in terms of overall ease of makeup and finished product attriisutes was obtained from Test Dough #5 (50 ppm Endo-H + 50 ppm cellulase preparation) . Example 7 Endo-H is evaluated at 800 ppm in a chemically leavened whole wheat cracker formula set forth in Table 3 below. The Test dough formula in column 2 is the sameas theControldoughformulaincolumn1 except fortheadditionofEndo-H. Themixtimes are 4minutes. Thedoughwateris determinedbyfeel forproper viscosity. The water in the formula includes 20 milliliters of water usedto dissolvethe aπmoniumbicarbonate. Floortime (dough rest time) is from about 30-90 minutes at roam temperature and humidity in a stainless steel bowl, covered lightly with plastic wrap. Sheeting is done by hand with a rolling pin. The crackers are cut cut, dockered, and salted by hand. The crackers are baked at 385°F for 13 minutes. Following baking, oil is brushed on surfaces. The dough from Test formula 2, con-taining the Endo-H, is slightly more extensible than the Control dough (formula 1), allowing for easier sheeting. Thus, the crackers from Endo-H dough are more uniform in overall texture in --comparison to crackers from the control dough. Dough Ingredients Whole Wheat Pastry Flour Hard Whole Wheat Flour Water Wheat Gluten Honey Shortening Sodium Bicarbonate Ammonium Bicarbonate Salt Monocalcium Hiosphate Endo-H Conditions Dough Temperature pH Bake Time

Claims: WHAT IS CLAIMED IS: 1. AdoughmixturecomprisingaTypeIIendoglycosidasealoneorincombination with additional enzymes. 2. A dough mixture comprising at least about 10 ppm of a Type II endoglycosidase. 3. A dough mixture of Claim 2 wherein the Type II endoglycosidase is at least about 25 ppm. 4. The dough mixture of Claim 2 wherein the Type II endoglycosidase is f om about 25 ppm to about 100 ppm. 5. The dough mixture of Claim 2 wherein the Type II endoglycosidase is from about 50 ppm to about 100 ppm. 6. The dough mixture of Claim 2 wherein the Type II endoglycosidase is from about 25 ppm to about 1000 ppm. 7. ThedoughmixtureofClaim1whereintheTypeII endoglycosidase is selected from the group consisting of Endo-H, Endo-F, Endo-L, Endo-CI, Endo-CII, an Endo-F-Gal type and HJGase. 8. ThedoughmixtureofClaim7whereintheTypeII endoglycosidase is at least about 10 ppm Endo-H. 9. Thedoutghmixtureof Claim7 whereintheTypeII endoglycosidase is at least about 50 ppm Endo-H. 10. A bread or roll dough mixture comprising at least about 25 ppm Endo-H. 11. A bread or roll dough mixture comprising about 10 ppm to about 1000 ppm of a Type II endoglycosidase. 12. The dough mixture of Claim 1 wherein the additional enzyme is cellulase. 13. A method of improving the characteristics of a dough or a baked good comprising the steps of: a) mixingaTypeU endoglycosidasewithdoughingredients to formadough mixture, and b) baking said dough mixture to form a baked good. * 14. The method of Claim 13 wherein the Type H endoglycosidase is capable of cleaving all or part of a carbohydrate moiety from a glycoprotein. 15. ThemethodofClaim13 whereintheTypeIIendoglycosidasecomprisesEndo-H. 16. Themethodof Claim15whereinthe Endo-Hcomprises at leastabout 10 ppm. 17. The method of Claim 16 wherein the Endo-H comprises between about 25 ppm and about 1000 ppm. 18. ThemethodofClaim13 whereintheType II endoglycosidase is selectedfrom thegroup consistingofEndo-F, Endo-L, Endo-C, Endo-dl, Endo-F-Galtype, and PNGase. 19. ThemethodofClaim13 whereintheTypeII endoglycosidase is selectedfrom the group consisting of endo-β -N-acetylglucosaminidase, endo-«-N- acelylgalac±csaminidase and endo-β-N-galactosidase. 20. ThemethodofClaim13whereincellulaseismixedwiththedoughingredients in step (a) . 21. Amethod of improvingbreador roll dough orbread orroll characteristics (Comprising the steps of: a) mixing a Type II endoglycosidasewithbread or roll dough ingredients to form a dough mixture; b) shaping or panning the dough mixture; c) proofping the dough mixture, and d) baking the dough mixture to form bread or rolls. 22. ThemethodofClaim21whereintheType II endoglycosidase is selectedfrom thegroup consistingofEndo-H, Endo-F, Endo-L, Endo-CI, Endo-CII, Endo-F- Gal type, Endo-F, and INGase. 23. Amethod of improving bread or roll dough orbread or roll characteristics ccπprising the steps of: a) mixingdoughingredientswithaType II endoglycosidaseto formadough mixture; b) allowing the dough mixture of step (a) to rest for between about 1 to 20 minutes at about 4°C to 40°C; c) shaping or panning the dough mixture; d) proofing the dough mixture a second time for between about 1 to about 90 minutes, and e) baking the dough mixture to form bread or rolls. 24. A method of iitproving dough characteristics comprising the steps of: a) mixing a Type II endoglycosidase with a first portion of dough ingredients to form a sponge; b) allowing the sponge to ferment; c) mixing the sponge from step (b) with a second portion of dough ingredients to form a dough mixture, and d) baking the dough mixture to form bread or rolls. 25. A method of improving dough characteristics cαπprising the steps of: a) mixing a Type II endoglycosidase with a first portion of dough ingredients to form a sponge; b) allowing the sponge to ferment; c) mixing the sponge from step (b) with a second portion of dough ingredients to form a dough mixture; d) shaping or panning the dough mixture, and e) baking the dough mixture to form bread or rolls. 26. Amethod ofijtrprovingbreaddoughcharacteristics comprisingthe steps of: a) mixing a first portion of dough ingredients to form a sponge; b) alloWping the sponge to ferment; c) mixing the sponge with a Type II endoglycosidase and a second portion of dough ingredients to form a dough mixture; d) shaping or panning the dough mixture from step (c) , and e) baking the dough mixture to form bread or rolls. 27. The method of Claim 26 wherein the Type II endoglycosidase is Endo-H. 28. Themethod of Claim 27 whereinthe Endo-H is at least about 25 ppm in the dough mixture. 29. Amethod of iπprovingbreaddoughcharacteristics comprisingthe steps of: a) mixing a first portion of dough ingredients to form a sponge; b) allowing the sponge to ferment; c) mixingthe spongewith aType II endoglycosidase and a secondportion of dough ingredients to form a dough mixture; d) shaping or panning the dough mixture from step (c) ; e) proofing the dough mixture, and f) baking the dough mixture to form bread or rolls.

Inventors: LAD PUSHKARAJ J, MULLINS M MARGARET, LAD, PUSHKARAJ, J., MULLINS, M., MARGARET
Applicants: GENENCOR INT, GENENCOR INTERNATIONAL, INC.
Application year: 1992
Main classification: A21D8
IPC codes: A21D8