Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
  • C12P7/00—Preparation of oxygen-containing organic compounds
  • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
  • C12P7/6436—Fatty acid esters
  • C12P7/6445—Glycerides
  • C12P7/6463—Glycerides obtained from glyceride producing microorganisms, e.g. single cell oil
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
  • A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
  • A23J1/18—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from yeasts
• • C—CHEMISTRY; METALLURGY
  • C10—PETROLEUM, GAS OR COKE INDUSTRIES; TECHNICAL GASES CONTAINING CARBON MONOXIDE; FUELS; LUBRICANTS; PEAT
  • C10G—CRACKING HYDROCARBON OILS; PRODUCTION OF LIQUID HYDROCARBON MIXTURES, e.g. BY DESTRUCTIVE HYDROGENATION, OLIGOMERISATION, POLYMERISATION; RECOVERY OF HYDROCARBON OILS FROM OIL-SHALE, OIL-SAND, OR GASES; REFINING MIXTURES MAINLY CONSISTING OF HYDROCARBONS; REFORMING OF NAPHTHA; MINERAL WAXES
  • C10G32/00—Refining of hydrocarbon oils by electric or magnetic means, by irradiation, or by using microorganisms
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
  • C12N1/005—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor after treatment of microbial biomass not covered by C12N1/02 - C12N1/08
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
  • C12N1/02—Separating microorganisms from their culture media
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
  • C12N1/26—Processes using, or culture media containing, hydrocarbons
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10S435/00—Chemistry: molecular biology and microbiology
  • Y10S435/8215—Microorganisms
  • Y10S435/822—Microorganisms using bacteria or actinomycetales
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10S435/00—Chemistry: molecular biology and microbiology
  • Y10S435/8215—Microorganisms
  • Y10S435/911—Microorganisms using fungi
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10S435/00—Chemistry: molecular biology and microbiology
  • Y10S435/8215—Microorganisms
  • Y10S435/911—Microorganisms using fungi
  • Y10S435/921—Candida
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10S435/00—Chemistry: molecular biology and microbiology
  • Y10S435/8215—Microorganisms
  • Y10S435/911—Microorganisms using fungi
  • Y10S435/921—Candida
  • Y10S435/923—Candida lipolytica
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10S435/00—Chemistry: molecular biology and microbiology
  • Y10S435/8215—Microorganisms
  • Y10S435/911—Microorganisms using fungi
  • Y10S435/933—Penicillium
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10S435/00—Chemistry: molecular biology and microbiology
  • Y10S435/8215—Microorganisms
  • Y10S435/911—Microorganisms using fungi
  • Y10S435/944—Torulopsis

Definitions

• the invention relates to a process for the production on hydrocarbons of edible micro-organisms in purified form, for example, yeast, bacteria of the orders of Pseudomonadales, Eub acteriales, Actinomycetales and Penicillium expansum.
• yeasts on hydrocarbons and in particular on petroleum hydrocarbons, as the carbonaceous substratum, generally gives yeasts which after washing and drying possess a peculiar taste which is sharp and rancid and which may interfere with their use in foods for human consumption.
• the traditional yeasts, cultivated on molasses, residuary liquors or bisulphite liquors also have a peculiar taste, different from that of yeasts cultivated on hydrocarbons, and certain processes have been proposed for attenuating this tastenotably to attenuate their bitterness, but none of these has succeeded completely.
• yeasts cultivated with hydrocarbons as the sole source of carbon have a distinctly higher lipid content than yeasts cultivated on the traditional substrate: molasses, residuary liquors, bisulphite liquors.
• Lipids consist essentially of fatty acids, esters (with greater or lesser degrees of oxidation) including fats and sterols. It has now been found that extraction of these lipids, wholly or in part, leads to a substantial reduction in this characteristic taste of yeasts cultivated on hydrocarbons or to the elimination of this taste.
• a process which comprises cultivating a microorganism in the presence of a feedstock consisting wholly or in part of straight chain hydrocarbons, separating from the product a fraction comprising the micro-organism and extracting said fraction by means of a solvent.
• Solvents which may be employed include ethyl alcohol, isopropanol, light hydrocarbons, including benzene and light platformate fractions, ethyl ether, acetone, chlorinated solvents and liquefied petroleum gases, such as butane and propane.
• micro-organism used herein we'include mixtures of micro-organisms.
• Micro-organisms which are cultivated as herein described may by yeasts, moulds or bacteria.
• a yeast is employed this is of the family Cryptococcaceae and particularly of the sub-family Cryptococcoideae; however, if desired there may be used, for example, ascosporogeneous yeasts of the sub-family Saccharomycoideae.
• Preferred genera of the Cryptococcoideae sub-family are Torulopsis (also known as Torula), Candida and Mycoderma.
• Preferred strains of yeast are as follows. In particular it is preferred to use the specific stock of indicated reference number;
• an aqueous nutrient medium and a supply of oxygen preferably in the form of air.
• a suitable nutrient medium for yeasts (and moulds) has the composition:
• a genus in the Actinomycetales order has the following composition:
• Another suitable nutrient medium for the growth of bacteria has the composition:
• the micro-organism will usually grow initially at a low rate of increase in cellular density. (This period of growth is referred to as the lag phase) Subsequently the rate of growth will increase to a higher rate of growth; the period at the higher rate of growth is referred to as the exponential phase and subsequently again the cellular density will become constant (the statitonary phase).
• a supply of the micro-organism for starting the next batch will preferably be removed before the termination of the exponential phase.
• the growth operation will usually be discontinued before the stationary phase.
• the micro-organism will usually be separated f'rom the bulk of the aqueous nutrient medium and from the bulk of the unused feedstock fraction.
• the growth of the micro-organism used is favoured by the addition to the culture medium of a very small proportion of extract of yeast (an industrial product rich in vitamins of group B obtained by the hydrolysis of a yeast) or more generally of vitamins of group B and/ or biotin.
• This quantity is preferably of the order of 25 parts per million with reference to the aqueous fermentation medium. It can be higher or lower according to the conditions chosen for the growth.
• the growth of the microorganism takes place at the expense of the feedstock fraction with the intermediate production of bodies having an acid function, principally fatty acids, in such manner that the pH of the aqueous mineral medium progressively diminishes. If one does not correct it the growth is fairly rapidly arrested and the concentration of the micro-organism in the medium, or cellular density, no longer increasees so that there is reached a so-called stationary phase.
• the aqueous nutrient medium is maintained at a desired pH by the step-Wise or continuous addition of an aqueous medium of high pH value.
• an aqueous medium of high pH value usually, when using moulds or yeasts and in particular when using Candida lipolytica, the pH of the nutrient medium will be maintained in the range 3-6 and preferably in the range 4-5. (Bacteria require a higher pH usually 6.5-8.)
• Suitable alkaline materials for addition to the growth mixture include sodium hydroxide, potassium hydroxide, disodium hydrogen phosphateand am: nionia, either free or in aqueous solution.
• the optimum temperature of the growth mixture will vary according to the type of micro-organism employed and will usually lie in the range 2535 C. When using Candida lipolytica the preferred temperature range is 28- 32 C.
• the take-up of oxygen is essential for the growth of the micro-organism.
• the oxygen will usually be provided as air.
• the air, used to provide oxygen should be present in the form of fine bubbles under the action of stirring.
• the air may be introduced through a sintered surface. However there may be used the system of intimate aeration known as vortex aeration.
• the micro-organism is preferably separated from the bulk of the liquid phase when possible by centrifuging and may be recovered as a cream or paste. However, in some cases separation will be accomplished by filtration or to some extent by decantation.
• This cream or paste which contains aqueous material may be treated by continuous solvent extraction or by successive washings with solvent followed by phase separation.
• the extraction is carried out in a stationary vessel equipped with paddle stirrer, preferably rotating at less than 10 revs per minute or in a vessel which rotates on a horizontal axis.
• the extract is withdrawn continuously and distilled, continuously or batchwise, at atmospheric or reduced pressure, and solvent continuously fed back to the extractor. troduced and withdrawn continuously or batchwise.
• the solvent extraction is effected while feeding solvent to the extractor at a periodically varying rate to create pulsations in the flow of said liquid stream.
• a device which imparts to it pulses whose amplitude and frequency are regulated experimentally at the most favourable value for each particular case.
• pulses are produced by any suitable processes already known, and preferably an alternating pump is used whose valves have been removed.
• the number of pulses lies between 1 and 60 per minute.
• the operation of the process under the action of pulses is further described in British patent application 2,234/ 63.
• a first extraction stage can be operated using a polar solvent, for example an alcoholic solvent, for example ethanol or isopropanol and then the partially purified micro-organism can be further treated in a second extraction stage using a hydrocarbon solvent, for example normal hexane or a light platformate fraction or benzene.
• a polar solvent for example an alcoholic solvent, for example ethanol or isopropanol
• a hydrocarbon solvent for example normal hexane or a light platformate fraction or benzene.
• solvent a mixture. of hydrocarbon in major amount with a polar solvent in minor amount.
• azeotropic mixture of hexane with isopropanol or ethanol is used. If desired both extraction stages can be operated in continuous manner.
• the yeast may be in- By the use of any alcohol in the first stage of a two stage extraction as hereinbefore described, the water content of the micro-organism cream or paste is considerably reduced. As a result the micro-organism containing material which is fed to the second stage has a sufficiently low content of water to ensure that the non-aqueous contaminants which are still present are miscible with the solevnt used in the second extraction stage.
• Each extraction stage may consist of either one or more substages consisting of washing with the solvent used in the stage followed by separation.
• the amount of ethanol or isopropanol which is used should be 1.5-3 times the volume of water which is present in the cream or paste of the micro-organism.
• two washings with ethanol or isopropanol may be employed using in the first washing a volume of solvent equal to the volume of water in the cream or paste and in the second washing a smaller amount of ethanol or isopropanol for example, one half of the amount used in the first washing.
• the cream or paste is allowed to drain, for example by filtering and part of the residual solvent is then preferably removed by vacuum filtration.
• the amount of solvent used in the (or each) washing will usually be 2-20 times the volume of the resulting dry micro-organism.
• the final stage employed for the removal of solvent is evaporation, suitably under reduced pressure and suitably in a stream of inert gas, for example, nitrogen or super-heated steam.
• inert gas for example, nitrogen or super-heated steam.
• the composition of the second stage solvent which will in any case acquire polar solvent from the first stage ex-' traction, can be stabilised.
• Build up of polar solvent can be avoided in the course of a distillation stage, in which the second stage solvent is recovered by the removal of separate streams consisting of (a) polar solvent for recycle to the first extraction stage and (b) a mixture of hydrocarbon and polar solvent for recycle to the second stage.
• the extract obtained by the second extraction stage is distilled to recover (a) overhead a mixture of hydrocarbon, polar solvent and water for recycle to the second extraction stage and (b) a bottoms fraction containing polar solvent, water and the extracted materials; this fraciton is preferably blended with the extract obtained in the first extraction stage before this is fed to distillation whereby all contaminants recovered by solvent extraction are removed as a bottoms fraction in this distillation stage.
• the polar solvent is ethanol or isopropanol.
• the second stage solvent is an azeotropic mixture.
• Optimum contact time will usually vary inversely with the temperature of extraction. It will usually be undesirable to use a temperature above 70 C. since higher temperatures will lead to some degradation of the product.
• the cream or paste of micro-organism is subjected to partial drying before solvent extraction it will then usually be possible to operate the first extraction stage with only a single washing with an alcoholic solvent and with the use of a smaller amount of solvent than would be required if no drying had taken place. If the extent of drying is considerable a first extraction stage using a polar solvent, for example an alcoholic solvent, is not necessary; in this case the single stage extraction process can be operated by the use of a solvent which is entirely hydrocarbon or which is a mixture of hydrocarbon and polar solvent, for example an alcohol or ketone or chlorinated hydrocarbon.
• a yeast which has been freed from the whole or part of its lipids and the contaminating hydrocarbons by one of the methods described herinbefore and whose taste has been improved is a new industrial product of value for human nutrition.
• the lipid extract which has been recovered by the evaporation of the solvent is also a new industrial product which can be used either as such or as a raw material for the separation of its sterols, fatty acids (either before saponification or after) or of its other constituents.
• EXAMPLE 1 A yeast cream of the strain Candida lipolytica was prepared as described in copending US. application S.N. 243,961 with reference to the diagram which accompanied said specification. It will be understood that the material used in the present example is the yeast cream obtained after washing with surface active yeast and with water and prior to the drying stage described with reference to said diagram.
• Yeast cream containing 2 parts by Weight of dry yeast and 8 parts by weight of water was charged to the extractor with 16 parts of ethanol. The mixture was maintained at 60 C. for 30 minutes while rotating the drum.
• Solvent was drawn off, finally under vacuum. There was thus obtained a mixture of 2 parts of yeast and 2 parts of solvent which consisted of /3 water and /3 of ethanol, together with some remaining contaminants.
• EXAMPLE 2 The yeast cream described in Example 1 was continuously extracted in a drum having its axis vertical and having a paddle stirrer which rotated at 10 revs/minute. In separate runs the solvent was respectively ethanol; ethanol followed by normal hexane; and isopropanol.
• the extract was continuously distilled for the recovery of lipids and hydrocarbons and the solvent recycled to the extractor.
• solvent used in the first-mentioned solvent extraction is an alcohol.
• Extract, Wt. (grams) 72 200 100 By Way of comparison then is provided the following description of Experiments 1 and 2 which do not constitute operation according to the invention.
• Example 1 The process described in Example 1 was repeated except that the yeast employed was a yeast Saccharomyces cerevisial cultivated in the known manner on a substratum of beet molasses. It is found that no perceptible improvement in the characteristic taste of this yeast is obtained.
• a process which comprises cultivating a straight chain hydrocarbon-consuming micro-organism selected from the group consisting of yeasts, bacteria of the orders Pseudomonadales, Eubacteriales and Actinomycetales; and Penicillium expansum in the presence of a feedstock consisting at least in part of a straight chain hydrocarbon, separating from the treated feedstock a fraction comprising the micro-organism and aqueous medium, reducing the proportion of water in said fraction by solvent extraction using a polar solvent and thereafter subjecting the raifinate, containing the micro-organism and a reduced proportion of aqueous medium so obtained to solvent extraction with a solvent containing in major amount a hydrocarbon the aforementioned solvent extraction.
• a straight chain hydrocarbon-consuming micro-organism selected from the group consisting of yeasts, bacteria of the orders Pseudomonadales, Eubacteriales and Actinomycetales; and Penicillium expansum in the presence of a feedstock consisting at least in part
• yeast is Candida lipolytica.

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Cited By (15)

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Citations (5)

• 1962
  • 1962-08-03 FR FR906076A patent/FR1340225A/fr not_active Expired
• 1963
  • 1963-06-25 GB GB18179/66A patent/GB1049066A/en not_active Expired
  • 1963-06-25 GB GB25210/63A patent/GB1049065A/en not_active Expired
  • 1963-06-25 GB GB18180/64A patent/GB1049067A/en not_active Expired
  • 1963-07-24 US US297213A patent/US3268419A/en not_active Expired - Lifetime
  • 1963-07-29 ES ES290663A patent/ES290663A1/es not_active Expired
  • 1963-08-02 BR BR151393/63A patent/BR6351393D0/pt unknown
• 1964
  • 1964-08-22 OA OA50285A patent/OA00220A/xx unknown
• 1968
  • 1968-01-08 CY CY42168A patent/CY421A/xx unknown
  • 1968-01-08 CY CY42268A patent/CY422A/xx unknown
  • 1968-01-08 CY CY42068A patent/CY420A/xx unknown
  • 1968-12-31 MY MY19686A patent/MY6800006A/xx unknown
  • 1968-12-31 MY MY196864A patent/MY6800064A/xx unknown
• 1969
  • 1969-12-31 MY MY1969222A patent/MY6900222A/xx unknown

Patent Citations (5)

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