CN104195180B - The method of fermenting and producing high density yeast extract emulsion - Google Patents
The method of fermenting and producing high density yeast extract emulsion Download PDFInfo
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- CN104195180B CN104195180B CN201410388212.1A CN201410388212A CN104195180B CN 104195180 B CN104195180 B CN 104195180B CN 201410388212 A CN201410388212 A CN 201410388212A CN 104195180 B CN104195180 B CN 104195180B
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Abstract
The invention discloses a kind of method of fermenting and producing high density yeast extract emulsion, its step is as follows：1）Saccharomyces cerevisiae slant strains are inoculated into molasses fluid nutrient medium, seed liquor is obtained；2）Seed liquor is inoculated into molasses culture medium, 28 DEG C of temperature intermittently sways culture；3）By step 2）Seed liquor access fermentation tank in, 4~6h of fermented and cultured adds dilution molasses and nutritive salt, 24~25h of stream plus culture by technological requirement mean flow afterwards；4）By step 3）Fermentation liquid added water centrifuge washing through centrifuge, obtain the yeast extract production emulsifiable paste of solid content 60%~70%, then the emulsifiable paste is diluted with water to the emulsion of concentration 10~20%.The present invention uses cane molasses for the production method of culture medium carbon source culture yeasts extract emulsion, shortens the production time, improves the level of production of yeast extract emulsion, and it is 250~300g/L to obtain yeast cells emulsion concentration, and protein content is up to 55%.
Description
Technical field
The invention belongs to yeast extract and biological fermentation field, and in particular to a kind of fermenting and producing high density extraction from yeast
The method of thing emulsion.
Background technology
Yeast extract is used using the food yeast of rich in protein as raw material, using self-dissolving, enzymolysis, separation, dense
The modern biotechnology new and high technologies such as contracting, refine after the protein in yeast cells, nucleic acid etc. are degraded and form, and are a kind of pale brown
Color solubility paste or slight yellow powdery natural product.Main component is polypeptide, amino acid, flavour nucleotide, B family vitamin
And trace element, it is the natural nutrition source for being especially suitable for microorganism growth, because yeast extract is nutritious, processing characteristics is good
It is good, tend to play the effectively delicious taste of enhancing product, mellow sense in some food processings, while product saline taste, tart flavour are relaxed,
Covering smell etc. is acted on, and is a kind of excellent natural flavouring, has in food service industry and have been widely used.
The raw material of production yeast extract is essentially from brewer's yeast and Saccharomyces cerevisiae, foreign countries' production yeast extract at present
Beer waste yeast is used mostly, and China is then main to produce yeast extract with Saccharomyces cerevisiae.The yeast produced with waste beer yeast
Extract is mainly the recycling of waste yeast, cheap, but can not develop high-end yeast extract product, such as high egg
In vain, high nucleic acid, high glutamic acid etc..More stable, the protein with the yeast extract quality that molasses fermented Saccharomyces cerevisiae is produced
High with nucleic acid content, the composition of nutriment is comprehensive.General high-end yeast extract product using professional Saccharomyces cerevisiae come
Production, yeast extract changes to the raw material based on Saccharomyces cerevisiae.
Latest data shows that in recent years, the average annual growth rate of demand of Chinese yeast extract is higher than more than 10%, and in U.S.
The growth rate of state and European market is about 5%.At present the whole world yeast extract year production scale at 200,000 tons or so.
China's production yeast extract faces insufficient raw material, and production technology falls behind relatively, and production cost is generally higher, product
Quality scale is relatively low to have a long way to go with the developed country such as American-European.
The raw material Patents and document report for retrieving production yeast extract at present are as follows：
1st, Chinese patent, a kind of Yeast extract with high glutamic acid content and preparation method thereof application number：CN200810189433.0
Open (bulletin) number：CN101756151A applies for (patent right) people：Angel Yeast Co., Ltd makes a summary：The present invention is carried
A kind of yeast extract and preparation method thereof is supplied, this method includes the yeast culture with a kind of protein content more than 60%
Cultivated for raw material in the culture medium rich in nitrogen source and yeast milk is made, a certain amount of protease and glutamine are then turned into ammonia
Enzyme is added in the yeast milk, so that a kind of yeast extract is made by series of process.Paddy in the yeast extract
Histidine content is more than 10%, the preparation method provided according to the present invention, and content of glutamic acid can be made on a large scale more than 10%
Yeast extract.
2nd, Chinese patent, the Application way application number of the extraction residue of yeast extract：CN201280006932.6, it is open
(bulletin) number：CN103338659A applies for (patent right) people：Xing Ren life sciences Co., Ltd. makes a summary：Obtain a kind of be applicable
In functional food or medicine etc., stay in grade, eliminate the glucose ceramide compositions of the impurity such as sterol glycocide.Make
For its raw material, using have edible experience, safe and easy purchase raw material, combined with inexpensive next life malaga glycosyl ceramide
Thing.The yeast residue after yeast extract will be extracted from torula etc. as raw material, it is carried out using organic solvents such as alcohols
Extract, so as to obtain the composition containing glucose ceramide.
3rd, Chinese patent, a kind of method for preparing yeast extract and β -1,3- glucans simultaneously, application number：
CN201010214448.5 is disclosed (bulletin) number：CN101897431A applies for (patent right) people：China Agricultural University makes a summary：
It is a kind of belong to yeast extract and beta-1,3-dextran preparation field while prepare yeast extract and beta-1,3-dextran
Method.This method includes the de- bitter removal of impurities of beer waste yeast, the enzymolysis of beer waste yeast and inactivation, the separation of yeast extract with
Dry, soda acid processing, washing and the drying and other steps of β -1,3- glucans.This method have raw material availability height, quality better, into
This is low, the features such as be easy to industrialization, realizes beer waste yeast recycling, and produce yeast extract and β -1,3- Portugals simultaneously
The purpose of two kinds of products of glycan.
4th, Chinese patent, the method application number of the mutton powder in non-meat source is prepared by de-fatted soybean dregs：
CN200810140581.3 is disclosed (bulletin) number：CN101322545A applies for (patent right) people：Henan capital food science and technology
Development corporation, Ltd. makes a summary：The method that the mutton powder in non-meat source is prepared by de-fatted soybean dregs, effectively solves non-meat source mutton powder
Production, to meet many-sided needs of problems, its technical scheme realized is, first by de-fatted soybean dregs, yeast extract and jade
The pure natural raw material in rice starch non-meat source is concentrated in twin-screw extruder, degraded, being reset and pyrolysis, is obtained
The expanded solid of micropore shape, then the expanded solid of micropore shape in leaching mode is equipped with other bastes, compound amino acid composition, use
Continuation mode is dried into belt drying machine, and non-meat source meat face powder is then made, further with other components, so as to
The flavorings such as the mutton powder of various local flavors are made, the present invention is the substitute of meat source meat face powder, and production capacity is high, cost is low, albumen contains
Amount is high, fragrance aromatic flavour is true to nature, can form the broad harmonious meat flavor from stewing to roasting, there is good economic and society
Benefit.
5th, Chinese patent, the method that zymosan, trehalose and yeast extract are prepared with beer waste yeast, application number：
CN200910036820.5, open (bulletin) number：CN101481719A, applies for (patent right) people：South China Science ＆ Engineering University;Guangzhou
Zhujiang River beer limited company makes a summary：One kind prepares zymosan, trehalose and extraction from yeast simultaneously using beer waste yeast
The method of thing.This method includes the de- bitter decontamination of beer waste yeast, the self-dissolving of beer waste yeast and enzymolysis and inactivation, and yeast is more
The separation and drying of sugar, the UF membrane of trehalose, concentration, crystallization and drying, the concentration of yeast extract and drying and other steps.Should
Method has raw material availability height, quality better, cost is low, the characteristics of be easy to industrialization, realizes from same industrial waste
Thing --- beer waste yeast produces zymosan, three kinds of products of trehalose and yeast extract simultaneously, improves industrial waste
The utilization rate of thing, has reached the purpose of discarded object maximum resource.
6th, Chinese patent, a kind of method application number that polypeptide and amino acid are extracted from beer waste yeast：
CN201110426167.0 is disclosed (bulletin) number：CN102550802A applies for (patent right) people：The grand peace biotechnology in Chengdu has
Limit company makes a summary：A kind of method that polypeptide and amino acid are extracted from beer waste yeast, beer waste yeast powder is preprocessed, height
Pressure homogenization, for the first time extract, digest, extracts for the second time, go out enzyme and press filtration separation after, be spray-dried obtain yeast polypeptides with
Amino acid powder.The present invention using brewery produce beer after accessory substance --- beer waste yeast as raw material, extract yeast polypeptides and
Amino acid powder, with pollution abatement and can realize the secondary conversion of resource, can also produce big economic benefit；The present invention is used
It is first high-pressure homogeneous, then extract twice, centre is aided with compound protease enzymolysis, can increase the permeability of yeast cell wall and carry
The dissolution rate of albumen and amino acid in high yeast cells, and then the pick-up rate of yeast extract is improved, increase containing for Yeast protein
Amount；The present invention can be used as functional feedstuff additive and food additives.
7th, Chinese periodical, the research Weng Xueqing of L-phenylalanine engineered strain high density fermentation, Fujian Mai Dan biologies group
Co., Ltd Foochow research center, fermentation science and technology communication 2012, summary:High density fermentation is one of Modern Fermentation Engineering new
Technology, can significantly improve the yield of colibacillus engineering body and gene engineering product.Fermentation for recombination bacillus coli comes
Say, realize high density fermentation, the volume of bioreactor and the separation expense of reduction biomass can be correspondingly reduced, so as to reduce
Production cost, reaches the purpose for improving production efficiency.Herein by the design of uniform Optimal Experimental to L-phenylalanine Escherichia coli
Research is optimized in the main component of engineering bacteria seed culture medium, and characterizes optimization by 30L automatic fermenters, as a result
Display：Ammonium sulfate 3.0g/L, magnesium sulfate 7.0g/L, yeast extract 13g/L have the synchronism of preferable engineering bacteria growing, plant
Sub- OD6100.190 is reached after diluting 100 times, is further to realize that high density fermentation production lays the first stone.
8th, Chinese periodical, the research of high-concentration culturing YE emulsions, Deng Chunming Chen Jia spring Ma Xingzhou, Guangxi light industry
2009, Guangdong Dan Baoli Yeast Co., Ltd, summary:In the culture of yeast extract emulsion, by adding in cane molasses stream
A certain proportion of fruit glucose syrup is added in liquid glucose to reduce the osmotic pressure of zymotic fluid, mitigates hyperosmosis to yeast cell growth
Inhibitory action, to reach the purpose of high-concentration culturing yeast cells.By optimization of orthogonal test technological condition for fermentation, in experiment
In achieve effect of the Yeast Cultivation at concentrations up to 320g/L.
By document finding disclosed above, the emulsion for producing yeast extract is generally beer waste yeast and a small amount of bread ferment
It is female.There is bitter taste more brewer's yeast emulsion, removal completely is difficult to during production yeast extract.And high density fermentation production bread ferment
The report that mother is used to produce yeast extract is few.
The content of the invention
The invention aims to solve the problem of above-mentioned prior art is present there is provided one kind using Saccharomyces cerevisiae and with
Sugar refinery molasses are the method that culture medium produces high density yeast extract emulsion.
The method of fermenting and producing high density yeast extract emulsion of the present invention, comprises the following steps：
a）Saccharomyces cerevisiae slant strains are inoculated in 100mL molasses liquid seed culture mediums, at 28 DEG C of temperature, interval
Sway culture 24h or slow-speed of revolution 100rpm shaken cultivations 24h.
b）By step a）Seed liquor 100mL be seeded to the 3L equipped with molasses liquid seed culture medium by 10% inoculum concentration
In triangular flask, 1000mL seed liquors are prepared.At 28 DEG C of temperature, culture 24h or slow-speed of revolution 100rpm shaken cultivations are intermittently swayed
24h。
c）By step b）Seed liquor by 15% inoculum concentration under the flame ring aseptic inoculation to being equipped with fermentation medium
In 15L fermentation tanks so that initial incubation liquid cumulative volume about 6200mL.Adjust 28 DEG C of temperature, pH3.5~4.5,0.1~0.3vvm
A small amount of aerlbic culture 4h~6h.Start to dilute molasses and nutritive salt, stream plus condition of culture by technological requirement flow feeding afterwards：Training
It is 28 DEG C to support temperature, pH3.5~4.5, and feed supplement stream adds 24h~25h under 1.0~2.0vvm of throughput.
d）By step c）The fermentation liquid of acquisition is through centrifuge 5000r/min, centrifuge washing 10min, and add water centrifuge washing
3~5 times, the yeast extract yeast cake emulsifiable paste of solid content 60%~70% is obtained, then the yeast cake emulsifiable paste is diluted with water
To 10~15% emulsion.
Wherein, step a）With step b）In molasses liquid seed culture medium formula be：Dilute molasses 150mL/L, urea
2.50g/L, MAP 0.60g/L, epsom salt 0.12g/L, white vitriol 0.05g/L, potassium chloride 0.10g/L, chlorine
Change sodium 0.10g/L, micro- mother liquor 2mL.Medium pH is adjusted to 4.2,121 DEG C of high-temperature sterilization 20min.
Wherein, step c）In fermentative medium formula be：Molasses 200mL, urea 10.0g, MAP 9.0g are diluted,
Epsom salt 7.5g, white vitriol 1.0g, potassium chloride 5g, sodium chloride 5g, micro- mother liquor 10mL.121 DEG C of high temperature go out
Bacterium 20min.
Wherein, described dilution molasses are that raw sugar honey is diluted with water to 45~50%oBx of brix, and 1min is boiled in heating, cools down
To 50~80 DEG C, centrifuge 4000r/min is used, 10min is centrifuged, discards sediment, produce dilution molasses.
Described stream Ensure Liquid salt is that urea 110g and MAP 20g, two kinds of nutritive salt are made into 500mL solution together.
121 DEG C of high-temperature sterilization 20min.
Wherein, described micro- mother liquor formula is：MnCI2·2H2O 4mg/L, KI 0.4 mg/L, pantothenic acid 4mg/
L, the mg/L of biotin 0.1, vitamin B10.4 g/L, the mg/L of vitamin C 4.
Wherein, step c）Described in dilute molasses and nutritive salt by technological requirement flow feeding：Add dilution sugar per tank stream
Honey about 5.0L, 20%, the 8h~18h mean flows for adding the sweet total amount of dilution by fed-batch cultivation time 0h~8h mean flows add dilution honey total
56%, 18h~24h mean flows of amount add the 24% of the sweet total amount of dilution.Nutritive salt urea 110g and MAP are needed per tank feed supplement
20g, two kinds of nutritive salt are made into 500mL solution together, by fed-batch cultivation time 0h~total liquid measure of 8h mean flow Ensure Liquid salt
The 50% of 50%, 8h~total liquid measure of 12h mean flow Ensure Liquid salt.
High density yeast extract emulsion produced by the invention, its protein content is high, can produce high-end yeast and take out
Extract product, effectively increases productivity effect.
Compared with the prior art, utstanding substantial characteristics and significant improvement of the present invention are：
1st, present aspect uses suitable fermentation process, rational culture medium prescription and fermentation method, obtain it is high containing protein,
The highdensity special production emulsion of yeast extract.
2nd, raw material sources enrich, and can select Saccharomyces cerevisiae, it would however also be possible to employ the yeast of molasses-spirit production process.
3rd, reaction condition is gentle,
4th, the extract obtained is in good taste, can improve the total nitrogen content and amino-acid nitrogen content in yeast extract product.
Brief description of the drawings
Accompanying drawing 1 is the schematic diagram of the production technology of the embodiment of the present invention.
See in figure, the method for fermenting and producing high density yeast extract emulsion of the invention, its step is as follows：1）By face
Bag yeast slant strains are inoculated into molasses fluid nutrient medium, obtain seed liquor；2）Seed liquor is inoculated into nutritive salt and molasses training
Support in base, 28 DEG C of temperature intermittently sways culture；3）By step 2）Seed liquor access 15L airlift fermentors in, fermented and cultured
4~6h, adds dilution molasses and nutritive salt, 24~25h of stream plus culture by technological requirement mean flow afterwards；4）By step 3）Fermentation
Mash adds water centrifuge washing through centrifuge, obtains the yeast extract production emulsifiable paste of solid content 60%~70%, then by the breast
Cream is diluted with water to the emulsion of concentration 10~20%.
Embodiment
Implement to be used to illustrate the present invention below, but be not limited to the scope of the present invention.
Example 1
The method of fermenting and producing high density yeast extract emulsion, is made up of following steps：
Step 1：Saccharomyces cerevisiae slant strains are inoculated in 100mL molasses liquid seed culture mediums, at 28 DEG C of temperature,
Have a rest and sway culture 24h.
Step 2：The seed liquor 100mL of step 1 is seeded to the 3L equipped with molasses liquid seed culture medium by 10% inoculum concentration
In triangular flask, 1000mL seed liquors are prepared.At 28 DEG C of temperature, culture 24h is intermittently swayed.
Wherein, the molasses liquid seed culture medium formula in step 1 and step 2 is：Dilute molasses 150mL/L, urea
2.50g/L, MAP 0.60g/L, epsom salt 0.12g/L, white vitriol 0.05g/L, potassium chloride 0.10g/L, chlorine
Change sodium 0.10g/L, micro- mother liquor 2mL.Medium pH is adjusted to 4.2,121 DEG C of high-temperature sterilization 20min.
Step 3：By the seed liquor of step 2 by 15% inoculum concentration under flame ring aseptic inoculation to be equipped with fermentation medium
15L fermentation tanks in so that initial incubation liquid cumulative volume about 6200mL.28 DEG C of temperature of regulation, pH3.5~4.5,0.1~
The a small amount of aerlbic culture 4h of 0.3vvm.Start flow feeding dilution molasses and nutritive salt afterwards.Stream plus condition of culture：Cultivation temperature is
28 DEG C, pH3.5~4.5, feed supplement stream adds 24h under 1.0~2.0vvm of throughput.
Wherein, the fermentative medium formula of 15L fermentation tanks is：Dilute molasses 200mL, urea 10.0g, MAP
9.0g, epsom salt 7.5g, white vitriol 1.0g, potassium chloride 5g, sodium chloride 5g, micro- mother liquor 10mL.It is 121 DEG C high
Temperature sterilizing 20min.
Micro- mother liquor formula：MnCI2·2H2O 4mg/L, KI 0.4 mg/L, pantothenic acid 4mg/L, biotin 0.1
Mg/L, vitamin B10.4 g/L, the mg/L of vitamin C 4.
Feeding medium during fermentation is formulated and feeding method control：Per tank stream add dilution molasses about 5.0L, by fed-batch cultivation time 0h~
8h mean flows add 20%, 8h~18h mean flows of the sweet total amount of dilution to add 56%, 18h~24h mean flows of the sweet total amount of dilution to add dilution
The 24% of sweet total amount.Nutritive salt urea 110g and MAP 20g are needed per tank feed supplement, it is molten that two kinds of nutritive salt are made into 500mL together
Liquid, by 50%, 8h~total liquid measure of 12h mean flow Ensure Liquid salt of fed-batch cultivation time 0h~total liquid measure of 8h mean flow Ensure Liquid salt
50%.Wherein, the dilution molasses and nutritive salt that stream adds all should 121 DEG C of high-temperature sterilization 20min.
Step 4：By fermentation liquid through centrifuge 5000r/min, centrifuge washing 10min, add water centrifuge washing 3~5 times, obtains
Solid content 60%~70% yeast extract yeast cake emulsifiable paste, then the emulsifiable paste is diluted with water to 10~15% emulsion.
As a result：Fermented and cultured terminates, and the barm cell concentration in fermentation liquid reaches 278.36g/L, and thalline size is equal
Even, gemma rate is below 1.0%, bacterium protein content 54.8%.
Example 2
The method of fermenting and producing high density yeast extract emulsion, is made up of following steps：
Step 1：Saccharomyces cerevisiae slant strains are inoculated in 100mL molasses liquid seed culture mediums, at 28 DEG C of temperature,
Have a rest and sway culture 24h.
Step 2：The seed liquor 100mL of step 1 is seeded to the 3L equipped with molasses liquid seed culture medium by 10% inoculum concentration
In triangular flask, 1000mL seed liquors are prepared.At 28 DEG C of temperature, culture 24h is intermittently swayed.
Wherein, the molasses liquid seed culture medium formula in step 1 and step 2 is：Dilute molasses 150mL/L, urea
2.50g/L, MAP 0.60g/L, epsom salt 0.12g/L, white vitriol 0.05g/L, potassium chloride 0.10g/L, chlorine
Change sodium 0.10g/L, micro- mother liquor 2mL.Medium pH is adjusted to 4.2,121 DEG C of high-temperature sterilization 20min.
Step 3：By the seed liquor of step 2 by 15% inoculum concentration under flame ring aseptic inoculation to be equipped with fermentation medium
15L fermentation tanks in so that initial incubation liquid cumulative volume about 6200mL.28 DEG C of temperature of regulation, pH3.5~4.5,0.1~
The a small amount of aerlbic culture 4h of 0.3vvm.Start flow feeding dilution molasses and nutritive salt afterwards.Stream plus condition of culture：Cultivation temperature is
28 DEG C, pH3.5~4.5, feed supplement stream adds 24h under 1.0~2.0vvm of throughput.
Wherein, the fermentative medium formula of 15L fermentation tanks is：Dilute molasses 200mL, urea 10.0g, MAP
9.0g, epsom salt 7.5g, white vitriol 1.0g, potassium chloride 5g, sodium chloride 5g, micro- mother liquor 10mL.It is 121 DEG C high
Temperature sterilizing 20min.
Micro- mother liquor formula：MnCI2·2H2O 4mg/L, KI 0.4 mg/L, pantothenic acid 4mg/L, biotin 0.1
Mg/L, vitamin B10.4 g/L, the mg/L of vitamin C 4.
Feeding medium during fermentation is formulated and feeding method control：Per tank stream add dilution molasses about 5.0L, by fed-batch cultivation time 0h~
8h mean flows add 20%, 8h~18h mean flows of the sweet total amount of dilution to add 56%, 18h~25h mean flows of the sweet total amount of dilution to add dilution
The 24% of sweet total amount.Nutritive salt urea 110g and MAP 20g are needed per tank feed supplement, it is molten that two kinds of nutritive salt are made into 500mL together
Liquid, by 50%, 8h~total liquid measure of 12h mean flow Ensure Liquid salt of fed-batch cultivation time 0h~total liquid measure of 8h mean flow Ensure Liquid salt
50%.Wherein, the dilution molasses and nutritive salt that stream adds all should 121 DEG C of high-temperature sterilization 20min.
Step 4：By fermentation liquid through centrifuge 5000r/min, centrifuge washing 10min, add water centrifuge washing 3~5 times, obtains
Solid content 60%~70% yeast extract yeast cake emulsifiable paste, then the emulsifiable paste is diluted with water to 10~15% emulsion.
As a result：Fermented and cultured terminates, and the barm cell concentration in fermentation liquid reaches 285.61g/L, and thalline size is equal
Even, gemma rate is below 1.0%, bacterium protein content 55.0%.
Example 3
The method of fermenting and producing high density yeast extract emulsion, is made up of following steps：
Step 1：Saccharomyces cerevisiae slant strains are inoculated in 100mL molasses liquid seed culture mediums, at 28 DEG C of temperature,
Have a rest and sway culture 24h.
Step 2：The seed liquor 100mL of step 1 is seeded to the 3L equipped with molasses liquid seed culture medium by 10% inoculum concentration
In triangular flask, 1000mL seed liquors are prepared.At 28 DEG C of temperature, culture 24h is intermittently swayed.
Wherein, the molasses liquid seed culture medium formula in step 1 and step 2 is：Dilute molasses 150mL/L, urea
2.50g/L, MAP 0.60g/L, epsom salt 0.12g/L, white vitriol 0.05g/L, potassium chloride 0.10g/L, chlorine
Change sodium 0.10g/L, micro- mother liquor 2mL.Medium pH is adjusted to 4.2,121 DEG C of high-temperature sterilization 20min.
Step 3：By the seed liquor of step 2 by 15% inoculum concentration under flame ring aseptic inoculation to be equipped with fermentation medium
15L fermentation tanks in so that initial incubation liquid cumulative volume about 6200mL.28 DEG C of temperature of regulation, pH3.5~4.5,0.1~
The a small amount of aerlbic culture 6h of 0.3vvm.Start flow feeding dilution molasses and nutritive salt afterwards.Stream plus condition of culture：Cultivation temperature is
28 DEG C, pH3.5~4.5, feed supplement stream adds 24h under 1.0~2.0vvm of throughput.
Wherein, the fermentative medium formula of 15L fermentation tanks is：Dilute molasses 200mL, urea 10.0g, MAP
9.0g, epsom salt 7.5g, white vitriol 1.0g, potassium chloride 5g, sodium chloride 5g, micro- mother liquor 10mL.It is 121 DEG C high
Temperature sterilizing 20min.
Micro- mother liquor formula：MnCI2·2H2O 4mg/L, KI 0.4 mg/L, pantothenic acid 4mg/L, biotin 0.1
Mg/L, vitamin B10.4 g/L, the mg/L of vitamin C 4.
Feeding medium during fermentation is formulated and feeding method control：Per tank stream add dilution molasses about 5.0L, by fed-batch cultivation time 0h~
8h mean flows add 20%, 8h~18h mean flows of the sweet total amount of dilution to add 56%, 18h~24h mean flows of the sweet total amount of dilution to add dilution
The 24% of sweet total amount.Nutritive salt urea 110g and MAP 20g are needed per tank feed supplement, it is molten that two kinds of nutritive salt are made into 500mL together
Liquid, by 50%, 8h~total liquid measure of 12h mean flow Ensure Liquid salt of fed-batch cultivation time 0h~total liquid measure of 8h mean flow Ensure Liquid salt
50%.Wherein, the dilution molasses and nutritive salt that stream adds all should 121 DEG C of high-temperature sterilization 20min.
Step 4：By fermentation liquid through centrifuge 5000r/min, centrifuge washing 10min, add water centrifuge washing 3~5 times, obtains
Solid content 60%~70% yeast extract yeast cake emulsifiable paste, then the emulsifiable paste is diluted with water to 10~15% emulsion.
As a result：Fermented and cultured terminates, and the barm cell concentration in fermentation liquid reaches 295.54g/L, and thalline size is equal
It is even, gemma rate 0.06%, bacterium protein content 55.2%.
Example 4
The method of fermenting and producing high density yeast extract emulsion, is made up of following steps：
Step 1：Saccharomyces cerevisiae slant strains are inoculated in 100mL molasses liquid seed culture mediums, at 28 DEG C of temperature,
Have a rest and sway culture 24h.
Step 2：The seed liquor 100mL of step 1 is seeded to the 3L equipped with molasses liquid seed culture medium by 10% inoculum concentration
In triangular flask, 1000mL seed liquors are prepared.At 28 DEG C of temperature, culture 24h is intermittently swayed.
Wherein, the molasses liquid seed culture medium formula in step 1 and step 2 is：Dilute molasses 150mL/L, urea
2.50g/L, MAP 0.60g/L, epsom salt 0.12g/L, white vitriol 0.05g/L, potassium chloride 0.10g/L, chlorine
Change sodium 0.10g/L, micro- mother liquor 2mL.Medium pH is adjusted to 4.2,121 DEG C of high-temperature sterilization 20min.
Step 3：By the seed liquor of step 2 by 15% inoculum concentration under flame ring aseptic inoculation to be equipped with fermentation medium
15L fermentation tanks in so that initial incubation liquid cumulative volume about 6200mL.28 DEG C of temperature of regulation, pH3.5~4.5,0.1~
The a small amount of aerlbic culture 6h of 0.3vvm.Start flow feeding dilution molasses and nutritive salt afterwards.Stream plus condition of culture：Cultivation temperature is
28 DEG C, pH3.5~4.5, feed supplement stream adds 24h under 1.0~2.0vvm of throughput.
Wherein, the fermentative medium formula of 15L fermentation tanks is：Dilute molasses 200mL, urea 10.0g, MAP
9.0g, epsom salt 7.5g, white vitriol 1.0g, potassium chloride 5g, sodium chloride 5g, micro- mother liquor 10mL.It is 121 DEG C high
Temperature sterilizing 20min.
Micro- mother liquor formula：MnCI2·2H2O 4mg/L, KI 0.4 mg/L, pantothenic acid 4mg/L, biotin 0.1
Mg/L, vitamin B10.4 g/L, the mg/L of vitamin C 4.
Feeding medium during fermentation is formulated and feeding method control：Per tank stream add dilution molasses about 5.0L, by fed-batch cultivation time 0h~
8h mean flows add 20%, 8h~18h mean flows of the sweet total amount of dilution to add 56%, 18h~25h mean flows of the sweet total amount of dilution to add dilution
The 24% of sweet total amount.Nutritive salt urea 110g and MAP 20g are needed per tank feed supplement, it is molten that two kinds of nutritive salt are made into 500mL together
Liquid, by 50%, 8h~total liquid measure of 12h mean flow Ensure Liquid salt of fed-batch cultivation time 0h~total liquid measure of 8h mean flow Ensure Liquid salt
50%.Wherein, the dilution molasses and nutritive salt that stream adds all should 121 DEG C of high-temperature sterilization 20min.
Step 4：By fermentation liquid through centrifuge 5000r/min, centrifuge washing 10min, add water centrifuge washing 3~5 times, obtains
Solid content 60%~70% yeast extract yeast cake emulsifiable paste, then the emulsifiable paste is diluted with water to 10~15% emulsion.
As a result：Fermented and cultured terminates, and the barm cell concentration in fermentation liquid reaches 300.08g/L, and thalline size is equal
Even, gemma rate 1.1%, bacterium protein content 55.0% makes flavouring raciness.
Claims (2)
1. a kind of method of fermenting and producing high density yeast extract emulsion, it is characterised in that comprise the following steps：
a）Saccharomyces cerevisiae slant strains are inoculated in 100mL molasses liquid seed culture mediums, at 28 DEG C of temperature, intermittently swayed
Cultivate 24h or slow-speed of revolution 100rpm shaken cultivations 24h；
b）By step a）Seed liquor 100mL be seeded to the 3L triangular flasks equipped with molasses liquid seed culture medium by 10% inoculum concentration
In, prepare 1000mL seed liquors；At 28 DEG C of temperature, culture 24h or slow-speed of revolution 100rpm shaken cultivations 24h is intermittently swayed；
c）By step b）Seed liquor by 15% inoculum concentration under flame ring aseptic inoculation arrive equipped with fermentation medium 15L hair
In fermentation tank so that initial incubation liquid cumulative volume about 6200mL；28 DEG C of temperature is adjusted, pH3.5~4.5,0.1~0.3vvm is a small amount of
Aerlbic culture 4h~6h；Start to dilute molasses and nutritive salt, stream plus condition of culture by technological requirement flow feeding afterwards：Culture temperature
Spend for 28 DEG C, pH3.5~4.5, feed supplement stream adds 24h~25h under 1.0~2.0vvm of throughput；
d）By step c）The fermentation liquid of acquisition is through centrifuge 5000r/min, centrifuge washing 10min, and add water centrifuge washing 3~5
It is secondary, the yeast extract yeast cake emulsifiable paste of solid content 60%~70% is obtained, then the yeast cake emulsifiable paste is diluted with water to 10
~15% emulsion；
Step a）With step b）In molasses liquid seed culture medium formula be：Dilute molasses 150mL/L, urea 2.50g/L, phosphorus
Acid one ammonium 0.60g/L, epsom salt 0.12g/L, white vitriol 0.05g/L, potassium chloride 0.10g/L, sodium chloride 0.10g/
L, micro- mother liquor 2mL, regulation medium pH to 4.2,121 DEG C of high-temperature sterilization 20min；
Step c）In fermentative medium formula be：Dilute molasses 200mL, urea 10.0g, MAP 9.0g, seven water sulfuric acid
Magnesium 7.5g, white vitriol 1.0g, potassium chloride 5g, sodium chloride 5g, micro- mother liquor 10mL, 121 DEG C of high-temperature sterilization 20min；
Described dilution molasses are that raw sugar honey is diluted with water to 45~50%oBx of brix, and heating boils 1min, is cooled to 50~80
DEG C, centrifuge 4000r/min is used, 10min is centrifuged, discards sediment, produce dilution molasses；
Step c）In stream Ensure Liquid salt be that urea 110g and MAP 20g, two kinds of nutritive salt are made into 500mL solution together,
121 DEG C of high-temperature sterilization 20min；
Step c）In stream add dilution molasses be through 121 DEG C of high-temperature sterilization 20min；
Described micro- mother liquor formula is：MnCI2·2H2O 4mg/L, KI 0.4 mg/L, pantothenic acid 4mg/L, biotin
0.1 mg/L, vitamin B10.4 g/L, the mg/L of vitamin C 4.
2. according to the method described in claim 1, it is characterised in that step c）Described in by technological requirement flow feeding dilution
Molasses and nutritive salt：Add dilution molasses about 5.0L per tank stream, add dilution molasses total amount by fed-batch cultivation time 0h~8h mean flows
20%, 8h~18h mean flows add dilution molasses total amount 56%, 18h~24h mean flows add dilution molasses total amount 24%；Per tank
Feed supplement needs nutritive salt to be urea 110g and MAP 20g, and two kinds of nutritive salt are made into 500mL solution together, by fed-batch cultivation
The 50% of 50%, 8h of time 0h~total liquid measure of 8h mean flow Ensure Liquid salt~total liquid measure of 12h mean flow Ensure Liquid salt.
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