Patent GB-1062005-A (GB)
Title: Improved process for biosynthesis and recovery of microbial cells
IPC: C12N1/26
Filed: 1965-06-08
Abstract: Protein-containing microbial cells produced in an aqueous biosynthesis bath comprising an inoculant of the micro-organism cells, a source of both carbon and hydrogen, and separate sources of oxygen and essential cell nutrients, are recovered by withdrawing an aqueous effluent (or aqueous froth product stream) containing the product microbial cells, adding to the effluent an essential cell nutrient-containing acid or a mixture of the acid with an essential cell nutrient-containing salt in sufficient amount to lower the pH of the effluent to a pH below that of the biosynthesis bath but above that at which the protein of the cells is degraded (preferably to between pH 2 and 6) - alternatively calcium chloride may be added alone without regard to the pH induced - heating the aqueous effluent to a temperature above the cell growth temperature but below that at which significant protein degradation occurs (preferably to between 45 DEG  and 100 DEG C) and thereafter separating the cells from the effluent to yield a cell component and a nutrient-containing medium. The separated cells may be heated as by spray-drying to render them non-viable. By the method bacterial cells of very small size e.g. from 0.5 to 5.0 microns and even smaller are coagulated whereby their separation from the effluent as by filtration, centrifugation and settling is facilitated.  The acidic nutrient-containing residium (preferably phosphorus-containing) is then returned to the biosynthesis bath and a basic nutrient-containing compound (preferably nitrogen-containing e.g. ammonium hydroxide) is added to the bath, either with the acidic residium or as a continuous direct addition, in amounts sufficient to maintain the pH of the bath at a suitable level for cell growth. The following species of micro-organisms are preferred for cultivation upon C1 to C30 hydrocarbon feeds containing less than 0.5 weight per cent of aromatics: Pseudomonas methanica, Nocardia corallina and Mycobacterium paraffinicum on C1 to C5 gaseous feeds; Pseudomonas fluorescens, P. desmolyticum and P. aeruginosa on C6 to C10 light naphtha feeds; Micrococcus cerificans, Pseudomonas aeruginosa, P. fluorescens, Norcardia opaca and N. rubra on C11 to C30 gas oils. Rhodotorula, Candida and Hansenula yeasts can also be employed. Alternatively, carbohydrate feeds may be used.  The nutrient-containing acid employed may be phosphoric acid or other specified phosphorus-containing acids; acetic, propionic or butyric acid; hydrochloric, sulphuric or nitric acid.  Suitable mixtures of nutrient-containing acids and salts include sodium and/or calcium chloride with phosphoric acid or acetic acid.  When the cultivation is performed for non-food purposes e.g. to produce intracellular and extracellular organic esters or acids, trichloro-acetic acid and the chlorides of zinc, tin and aluminium may be employed.  Suitable nitrogen-containing basic compounds include sodium nitrate, ammonia, ammonium hydroxide, ammonium nitrate, diammonium phosphate, urea, purine and asparagine. Examples are directed to the recovery of cells of Micrococcus cerificans ATCC 14987 cultivated in an aqueous biosynthesis bath containing 1.0 weight per cent of n-hexadecane.