CN104195180A - Method for fermentation production of high density yeast extract emulsion - Google Patents
Method for fermentation production of high density yeast extract emulsion Download PDFInfo
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Abstract
The invention discloses a method for fermentation production of a high density yeast extract emulsion. The method comprises the following steps: 1) inoculating a slant strain of baker's yeast into a molasses liquid culture medium to obtain a seed liquid; 2) inoculating the seed liquid into the molasses liquid culture medium and carrying out intermittent oscillating culture at 28 DEG C; 3) inoculating the seed liquid in the step 2) into a fermentation tank, carrying out fermentation culture for 4-6 hours, then, averagely feeding diluted molasses and nutritive salt according to process requirements, and carrying out feeding culture for 24-25 hours; and 4) adding water into fermentation mash by a virtue of a centrifugal machine to centrifugally wash to obtain yeast extract production emulsion paste with the content of solids of 60-70%, and then, adding water into the emulsion paste to dilute to obtain an emulsion with the using concentration of 10-20%. According to the production method which is used for culturing the yeast extract emulsion by using cane molasses as a carbon source of a culture medium, the production time is shortened and the production level of the yeast extract production emulsion is improved. The concentration of the yeast cell emulsion obtained is 250-300g/L and the content of proteins reaches up to 55%.
Description
Technical field
The invention belongs to yeast extract and biological fermentation field, be specifically related to a kind of method of fermentative production high-density yeast extract emulsion.
Background technology
Yeast extract adopts taking the food-yeast of rich in protein as raw material, adopt self-dissolving, enzymolysis, separation, the modern biological new and high technology such as concentrated, after protein, nucleic acid etc. in yeast cell are degraded, refining forming, is a kind of brown color solubility paste or light yellow powdery natural product.Main component is polypeptide, amino acid, flavour nucleotide, vitamin B group and trace element, it is the natural nutrition source that is applicable to very much microorganism growth, because yeast extract is nutritious, processing characteristics is good, in some food-processings, often can play the delicious taste of effective enhancing product, mellow sense, relax product saline taste, tart flavour simultaneously, the effects such as covering smell, are a kind of good natural flavourings, and in food service industry, tool has been widely used.
The raw material of producing yeast extract at present, mainly from cereuisiae fermentum and bread yeast, is produced yeast extract abroad and is mostly used beer waste yeast, and China mainly produces yeast extract with bread yeast.The yeast extract of producing with waste beer yeast is mainly the recycling of waste yeast, cheap, but cannot develop high-end yeast extract product, as high protein, high nucleic acid, high glutamic acid etc.The yeast extract quality of producing with molasses fermented bread yeast is more stable, and protein and nucleic acid content are high, and the composition of nutritive substance is comprehensive.General high-end yeast extract product adopts professional bread yeast to produce, and yeast extract is to changing taking bread yeast as main raw material.
Latest data shows, in recent years, the average annual growth rate of demand of yeast extract of China is higher than more than 10%, and in the rate of growth in US and European market greatly about 5%.Current global yeast extract year industrial scale 200,000 tons of left and right.
The developed countries such as China produces yeast extract and faces insufficient raw material, and production technology is relatively backward, and production cost is generally higher, and quality product class is relatively low and American-European have a long way to go.
The raw material Patents and the bibliographical information that retrieve current production yeast extract are as follows:
1, Chinese patent, a kind of Yeast extract with high glutamic acid content and preparation method thereof application number: CN200810189433.0 open (bulletin) number: CN101756151A application (patent right) people: Angel Yeast Co.,Ltd's summary: the invention provides a kind of yeast extract and preparation method thereof, the method comprises that exceed 60% yeast culture taking a kind of protein content cultivates and make yeast-lactic as raw material in the substratum that is rich in nitrogenous source, then a certain amount of proteolytic enzyme and glutamine transaminase are joined in described yeast-lactic, thereby make a kind of yeast extract through series of process.Content of glutamic acid in this yeast extract is greater than 10%, according to preparation method provided by the invention, can make on a large scale the yeast extract that content of glutamic acid is greater than 10%.
2, Chinese patent, the extraction residue of yeast extract utilize method application number: CN201280006932.6, open (bulletin) number: CN103338659A application (patent right) people: Xing Ren life science Co., Ltd. makes a summary: obtain a kind of glucose ceramide composition that is applicable to functional foodstuff or medicine etc., stay in grade, has removed the impurity such as sterol glycoside.As its raw material, the raw material that use has edible experience, safety and easily purchases, with low cost malaga in next life glycosyl ceramide composition.Using the yeast residue from the extraction yeast extracts such as torula as raw material, utilize the organic solvents such as alcohols to extract it, thereby obtain the composition that contains glucose ceramide.
3, Chinese patent, one is prepared yeast extract and β-1 simultaneously, the method of 3-dextran, application number: CN201010214448.5 open (bulletin) number: CN101897431A applies for (patent right) people: China Agricultural University's summary: one belongs to yeast extract and β-1, when the preparation field of 3-dextran, prepare yeast extract and β-1, the method for 3-dextran.The method comprises the removal of impurities of beer waste yeast debitterize, the enzymolysis of beer waste yeast and deactivation, the separation of yeast extract and dry, β-1, acid-alkali treatment, washing and the drying and other steps of 3-dextran.The method has that raw material availability is high, quality better, cost are low, be easy to the features such as industrialization, has realized beer waste yeast recycling, and has produced yeast extract and β-1 simultaneously, the object of two kinds of products of 3-dextran.
4, Chinese patent, prepared the method application number of the mutton powder in non-meat source: CN200810140581.3 open (bulletin) number: CN101322545A application (patent right) people by de-fatted soybean dregs: Henan Jinghua Food Science and Technology Co., Ltd.'s summary: the method for being prepared the mutton powder in non-meat source by de-fatted soybean dregs, efficient solution is the production of meat source mutton powder by no means, to meet many-sided needs of problems, the technical scheme of its realization is, first by de-fatted soybean dregs, the non-meat of yeast extract and W-Gum source pure natural raw material concentrates in twin-screw extruder, degraded, reset and pyrolysis, obtain the expanded solid of micropore shape, again expanded micropore shape solid is equipped with to other sauce to soak pouring mode, aminoacids complex composition, entering belt drying machine with continuous mode dries, then make non-meat source meat face powder, again further with other component, thereby can be made into the food flavourings such as the mutton powder of various local flavors, the present invention is the substitute of meat source meat face powder, production capacity is high, cost is low, protein content is high, fragrance aromatic flavour is true to nature, can form the meat flavor that boils the broad harmony of barbecue from stewing, there is good economic and social benefit.
5, Chinese patent; prepare the method for zymosan, trehalose and yeast extract with beer waste yeast; application number: CN200910036820.5, open (bulletin) number: CN101481719A, application (patent right) people: South China Science & Engineering University; Guangzhou Zhujiang Brewery Co., Ltd.'s summary: a kind of method of utilizing beer waste yeast simultaneously to prepare zymosan, trehalose and yeast extract.The method comprises beer waste yeast debitterize decon, the self-dissolving of beer waste yeast and enzymolysis and deactivation, the separation of zymosan and dry, the membrane sepn of trehalose, concentrated, crystallization and dry, the concentrated and drying and other steps of yeast extract.The feature that the method has that raw material availability is high, quality better, cost are low, be easy to industrialization, realize from same trade waste---beer waste yeast is produced zymosan, trehalose and three kinds of products of yeast extract simultaneously, improve the utilization ratio of trade waste, reached the object of waste maximum resource.
6, Chinese patent, one is extracted polypeptide and amino acid whose method application number: CN201110426167.0 open (bulletin) number: CN102550802A application (patent right) people from beer waste yeast: Chengdu HongAn Biotechnology Co., Ltd.'s summary: a kind of polypeptide and amino acid whose method extracted from beer waste yeast, beer waste yeast powder is after pre-treatment, high-pressure homogeneous processing, extracting for the first time, enzymolysis, extracting for the second time, the enzyme that goes out separate with press filtration, and spraying is dried and obtains yeast polypeptides and amino acid powder.The present invention produces the by product after beer taking brew-house---and beer waste yeast, as raw material, extracts yeast polypeptides and amino acid powder, can pollution abatement and realize the twice transformation of resource, also can produce large economic benefit; What the present invention adopted is first high-pressure homogeneous, twice extracting again, centre is aided with compound protease enzymolysis, can increase the permeability of yeast cells wall and improve albumen and amino acid whose solubility rate in yeast cell, and then the pick-up rate of raising yeast extract, the content of increase Yeast protein; The present invention can be used as functional feedstuff additive and foodstuff additive.
7, Chinese periodical, the research Weng Xueqing of L-Phe engineering strain high density fermentation, research centre, the biological Group Co.,Ltd Foochow of Fujian Mai Dan, the scientific and technological communication 2012 of fermenting, summary: high density fermentation is a new technology of Modern Fermentation Engineering, can significantly improve the output of colibacillus engineering body and gene engineering product.For the fermentation of recombination bacillus coli, realize high density fermentation, can correspondingly dwindle the separation costs of volume and the reduction biomass of bio-reactor, thereby reduce production costs, reach the object of enhancing productivity.By even optimization experiment design, the main component of L-Phe colibacillus engineering seed culture medium is optimized to research herein, and characterize optimization by 30L automatic fermenter, result shows: ammonium sulfate 3.0g/L, magnesium sulfate 7.0g/L, yeast extract 13g/L has the synchronism of good engineering bacteria growth, seed OD
610after diluting 100 times, reach 0.190, lay the first stone for further realizing high density fermentation production.
8, Chinese periodical, the research of high-concentration culturing YE emulsion, Deng Chunming Chen Jia spring Ma Xingzhou, Guangxi light industry 2009, Guangdong Dan Baoli Yeast Co., Ltd, summary: in the cultivation of yeast extract emulsion, reduce the osmotic pressure of fermented liquid by adding a certain proportion of fruit glucose syrup at cane molasses stream in liquid glucose, alleviate the restraining effect of high osmotic pressure to yeast cell growth, to reach the object of high-concentration culturing yeast cell.By optimization of orthogonal test technological condition for fermentation, obtain in test the effect of yeast culture concentration up to 320g/L.
By above-mentioned open source literature finding, mostly be beer waste yeast and a small amount of bread yeast for the production of the emulsion of yeast extract.Cereuisiae fermentum emulsion has bitter taste more, while producing yeast extract, is difficult to remove completely.And high density fermentation production bread yeast is few for the production of the report of yeast extract.
Summary of the invention
The object of the invention is the problem existing in order to solve above-mentioned prior art, a kind of bread yeast and method taking sugar refinery molasses as substratum production high-density yeast extract emulsion utilized is provided.
The method of fermentative production high-density yeast extract emulsion of the present invention, comprises the steps:
A). bread yeast slant strains is inoculated in 100mL molasses liquid seed culture medium, at 28 DEG C of temperature, intermittently sways and cultivate 24h or slow speed of revolution 100rpm shaking culture 24h.
B). the seed liquor 100mL of step a) is seeded in the 3L triangular flask that molasses liquid seed culture medium is housed to preparation 1000mL seed liquor by 10% inoculum size.At 28 DEG C of temperature, intermittently sway and cultivate 24h or slow speed of revolution 100rpm shaking culture 24h.
C). the seed liquor of step b) to being equipped with in the 15L fermentor tank of fermention medium, is made to the about 6200mL of initial incubation liquid cumulative volume by inoculum size aseptic inoculation under flame ring of 15%.Regulate 28 DEG C of temperature, pH3.5～4.5,0.1～0.3vvm is aerlbic culture 4h～6h in a small amount.Start afterwards by processing requirement flow feeding dilution molasses and nutritive salt, feeding culture condition: culture temperature is 28 DEG C, pH3.5～4.5, under air flow 1.0～2.0vvm, feed supplement stream adds 24h～25h.
D). the fermentation liquid that step c) is obtained is through whizzer 5000r/min, centrifuge washing 10min, add water centrifuge washing 3～5 times, obtains the yeast extract fresh yeast emulsifiable paste of solid content 60%～70%, then this fresh yeast emulsifiable paste is diluted with water to 10～15% emulsion.
Wherein, the molasses liquid seed culture medium formula in step a) and step b) is: dilution molasses 150mL/L, urea 2.50g/L, monoammonium phosphate 0.60g/L, magnesium sulfate heptahydrate 0.12g/L, Zinc Sulphate Heptahydrate 0.05g/L, Repone K 0.10g/L, sodium-chlor 0.10g/L, micro-mother liquor 2mL.Regulate medium pH to 4.2,121 DEG C of high-temperature sterilization 20min.
Wherein, the fermentative medium formula in step c) is: dilution molasses 200mL, urea 10.0g, monoammonium phosphate 9.0g, magnesium sulfate heptahydrate 7.5g, Zinc Sulphate Heptahydrate 1.0g, Repone K 5g, sodium-chlor 5g, micro-mother liquor 10mL.121 DEG C of high-temperature sterilization 20min.
Wherein, described dilution molasses are that former molasses are diluted with water to hammer degree 45～50%oBx, and heated and boiled 1min, is cooled to 50～80 DEG C, uses whizzer 4000r/min, and centrifugation 10min, discards throw out, obtains dilution molasses.
Described stream Ensure Liquid salt is, urea 110g and monoammonium phosphate 20g, and 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: MnCI
22H
2o 4mg/L, KI 0.4 mg/L, pantothenic acid 4mg/L, vitamin H 0.1 mg/L, vitamins B
10.4 g/L, vitamins C 4 mg/L.
Wherein, described in step c) by processing requirement flow feeding dilution molasses and nutritive salt: every tank stream adds the about 5.0L of dilution molasses, add fermentation time 0h～8h average flow by stream and add 20% of the sweet total amount of dilution, 8h～18h average flow adds 56% of the sweet total amount of dilution, and 18h～24h average flow adds 24% of the sweet total amount of dilution.Every tank feed supplement needs nutritive salt urea 110g and monoammonium phosphate 20g, and two kinds of nutritive salt are made into 500mL solution together, adds 50% of the total liquid measure of fermentation time 0h～8h average flow Ensure Liquid salt, 50% of the total liquid measure of 8h～12h average flow Ensure Liquid salt by stream.
The high-density yeast extract emulsion that the present invention produces, its protein content is high, can produce high-end yeast extract product, has effectively improved productivity effect.
Compared with the prior art, outstanding substantive distinguishing features of the present invention and significant progress are:
1, this aspect adopts suitable fermentation process, and reasonably culture medium prescription and fermentation mode obtains high containing protein, the special production emulsion of highdensity yeast extract.
2, raw material sources are abundant, can select bread yeast, also can adopt the yeast of molasses-spirit production process.
3, reaction conditions gentleness,
4, the extract mouthfeel obtaining is good, can improve 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 technique of the embodiment of the present invention.
In figure, see, the method for fermentative production high-density yeast extract emulsion of the present invention, its step is as follows: 1) bread yeast slant strains is inoculated in molasses liquid nutrient medium, obtains seed liquor; 2) seed liquor is inoculated in nutritive salt and molasses culture medium, 28 DEG C of temperature, intermittently sway cultivation; 3) by step 2) seed liquor access 15L airlift fermentor in, fermentation culture 4～6h, adds dilution molasses and nutritive salt, feeding culture 24～25h by processing requirement average flow afterwards; 4) by the fermentation liquid of step 3) through the whizzer centrifuge washing that adds water, the yeast extract that obtains solid content 60%～70% is produced emulsifiable paste, then this emulsifiable paste is diluted with water to the emulsion of working concentration 10～20%.
Embodiment
Below implement to be used for illustrating the present invention, but be not used for limiting the scope of the invention.
example 1
The method of fermentative production high-density yeast extract emulsion, is made up of following steps:
Step 1: bread yeast slant strains is inoculated in 100mL molasses liquid seed culture medium, at 28 DEG C of temperature, intermittently sways and cultivate 24h.
Step 2: the seed liquor 100mL of step 1 is seeded in the 3L triangular flask that molasses liquid seed culture medium is housed to preparation 1000mL seed liquor by 10% inoculum size.At 28 DEG C of temperature, intermittently sway and cultivate 24h.
Wherein, the molasses liquid seed culture medium formula in step 1 and step 2 is: dilution molasses 150mL/L, urea 2.50g/L, monoammonium phosphate 0.60g/L, magnesium sulfate heptahydrate 0.12g/L, Zinc Sulphate Heptahydrate 0.05g/L, Repone K 0.10g/L, sodium-chlor 0.10g/L, micro-mother liquor 2mL.Regulate medium pH to 4.2,121 DEG C of high-temperature sterilization 20min.
Step 3: the seed liquor of step 2 to being equipped with in the 15L fermentor tank of fermention medium, is made to the about 6200mL of initial incubation liquid cumulative volume by inoculum size aseptic inoculation under flame ring of 15%.Regulate 28 DEG C of temperature, pH3.5～4.5,0.1～0.3vvm is aerlbic culture 4h in a small amount.Start afterwards flow feeding dilution molasses and nutritive salt.Feeding culture condition: culture temperature is 28 DEG C, pH3.5～4.5, under air flow 1.0～2.0vvm, feed supplement stream adds 24h.
Wherein, the fermentative medium formula of 15L fermentor tank is: dilution molasses 200mL, urea 10.0g, monoammonium phosphate 9.0g, magnesium sulfate heptahydrate 7.5g, Zinc Sulphate Heptahydrate 1.0g, Repone K 5g, sodium-chlor 5g, micro-mother liquor 10mL.121 DEG C of high-temperature sterilization 20min.
Trace element mother liquor formula: MnCI
22H
2o 4mg/L, KI 0.4 mg/L, pantothenic acid 4mg/L, vitamin H 0.1 mg/L, vitamins B
10.4 g/L, vitamins C 4 mg/L.
Feeding medium during fermentation formula and feeding method control: every tank stream adds the about 5.0L of dilution molasses, add fermentation time 0h～8h average flow by stream and add 20% of the sweet total amount of dilution, and 8h～18h average flow adds 56% of the sweet total amount of dilution, and 18h～24h average flow adds 24% of the sweet total amount of dilution.Every tank feed supplement needs nutritive salt urea 110g and monoammonium phosphate 20g, and two kinds of nutritive salt are made into 500mL solution together, adds 50% of the total liquid measure of fermentation time 0h～8h average flow Ensure Liquid salt, 50% of the total liquid measure of 8h～12h average flow Ensure Liquid salt by stream.Wherein, the dilution molasses that stream adds and nutritive salt all should 121 DEG C of high-temperature sterilization 20min.
Step 4: by fermentation liquid through whizzer 5000r/min, centrifuge washing 10min, the centrifuge washing 3～5 times of adding water, obtains the yeast extract fresh yeast emulsifiable paste of solid content 60%～70%, then this emulsifiable paste is diluted with water to 10～15% emulsion.
Result: fermentation culture finishes, the barm cell concentration in fermentation liquid reaches 278.36g/L, and thalline size is evenly, and gemma rate is below 1.0%, bacterium protein content 54.8%.
example 2
The method of fermentative production high-density yeast extract emulsion, is made up of following steps:
Step 1: bread yeast slant strains is inoculated in 100mL molasses liquid seed culture medium, at 28 DEG C of temperature, intermittently sways and cultivate 24h.
Step 2: the seed liquor 100mL of step 1 is seeded in the 3L triangular flask that molasses liquid seed culture medium is housed to preparation 1000mL seed liquor by 10% inoculum size.At 28 DEG C of temperature, intermittently sway and cultivate 24h.
Wherein, the molasses liquid seed culture medium formula in step 1 and step 2 is: dilution molasses 150mL/L, urea 2.50g/L, monoammonium phosphate 0.60g/L, magnesium sulfate heptahydrate 0.12g/L, Zinc Sulphate Heptahydrate 0.05g/L, Repone K 0.10g/L, sodium-chlor 0.10g/L, micro-mother liquor 2mL.Regulate medium pH to 4.2,121 DEG C of high-temperature sterilization 20min.
Step 3: the seed liquor of step 2 to being equipped with in the 15L fermentor tank of fermention medium, is made to the about 6200mL of initial incubation liquid cumulative volume by inoculum size aseptic inoculation under flame ring of 15%.Regulate 28 DEG C of temperature, pH3.5～4.5,0.1～0.3vvm is aerlbic culture 4h in a small amount.Start afterwards flow feeding dilution molasses and nutritive salt.Feeding culture condition: culture temperature is 28 DEG C, pH3.5～4.5, under air flow 1.0～2.0vvm, feed supplement stream adds 24h.
Wherein, the fermentative medium formula of 15L fermentor tank is: dilution molasses 200mL, urea 10.0g, monoammonium phosphate 9.0g, magnesium sulfate heptahydrate 7.5g, Zinc Sulphate Heptahydrate 1.0g, Repone K 5g, sodium-chlor 5g, micro-mother liquor 10mL.121 DEG C of high-temperature sterilization 20min.
Trace element mother liquor formula: MnCI
22H
2o 4mg/L, KI 0.4 mg/L, pantothenic acid 4mg/L, vitamin H 0.1 mg/L, vitamins B
10.4 g/L, vitamins C 4 mg/L.
Feeding medium during fermentation formula and feeding method control: every tank stream adds the about 5.0L of dilution molasses, add fermentation time 0h～8h average flow by stream and add 20% of the sweet total amount of dilution, and 8h～18h average flow adds 56% of the sweet total amount of dilution, and 18h～25h average flow adds 24% of the sweet total amount of dilution.Every tank feed supplement needs nutritive salt urea 110g and monoammonium phosphate 20g, and two kinds of nutritive salt are made into 500mL solution together, adds 50% of the total liquid measure of fermentation time 0h～8h average flow Ensure Liquid salt, 50% of the total liquid measure of 8h～12h average flow Ensure Liquid salt by stream.Wherein, the dilution molasses that stream adds and nutritive salt all should 121 DEG C of high-temperature sterilization 20min.
Step 4: by fermentation liquid through whizzer 5000r/min, centrifuge washing 10min, the centrifuge washing 3～5 times of adding water, obtains the yeast extract fresh yeast emulsifiable paste of solid content 60%～70%, then this emulsifiable paste is diluted with water to 10～15% emulsion.
Result: fermentation culture finishes, the barm cell concentration in fermentation liquid reaches 285.61g/L, and thalline size is evenly, and gemma rate is below 1.0%, bacterium protein content 55.0%.
example 3
The method of fermentative production high-density yeast extract emulsion, is made up of following steps:
Step 1: bread yeast slant strains is inoculated in 100mL molasses liquid seed culture medium, at 28 DEG C of temperature, intermittently sways and cultivate 24h.
Step 2: the seed liquor 100mL of step 1 is seeded in the 3L triangular flask that molasses liquid seed culture medium is housed to preparation 1000mL seed liquor by 10% inoculum size.At 28 DEG C of temperature, intermittently sway and cultivate 24h.
Wherein, the molasses liquid seed culture medium formula in step 1 and step 2 is: dilution molasses 150mL/L, urea 2.50g/L, monoammonium phosphate 0.60g/L, magnesium sulfate heptahydrate 0.12g/L, Zinc Sulphate Heptahydrate 0.05g/L, Repone K 0.10g/L, sodium-chlor 0.10g/L, micro-mother liquor 2mL.Regulate medium pH to 4.2,121 DEG C of high-temperature sterilization 20min.
Step 3: the seed liquor of step 2 to being equipped with in the 15L fermentor tank of fermention medium, is made to the about 6200mL of initial incubation liquid cumulative volume by inoculum size aseptic inoculation under flame ring of 15%.Regulate 28 DEG C of temperature, pH3.5～4.5,0.1～0.3vvm is aerlbic culture 6h in a small amount.Start afterwards flow feeding dilution molasses and nutritive salt.Feeding culture condition: culture temperature is 28 DEG C, pH3.5～4.5, under air flow 1.0～2.0vvm, feed supplement stream adds 24h.
Wherein, the fermentative medium formula of 15L fermentor tank is: dilution molasses 200mL, urea 10.0g, monoammonium phosphate 9.0g, magnesium sulfate heptahydrate 7.5g, Zinc Sulphate Heptahydrate 1.0g, Repone K 5g, sodium-chlor 5g, micro-mother liquor 10mL.121 DEG C of high-temperature sterilization 20min.
Trace element mother liquor formula: MnCI
22H
2o 4mg/L, KI 0.4 mg/L, pantothenic acid 4mg/L, vitamin H 0.1 mg/L, vitamins B
10.4 g/L, vitamins C 4 mg/L.
Feeding medium during fermentation formula and feeding method control: every tank stream adds the about 5.0L of dilution molasses, add fermentation time 0h～8h average flow by stream and add 20% of the sweet total amount of dilution, and 8h～18h average flow adds 56% of the sweet total amount of dilution, and 18h～24h average flow adds 24% of the sweet total amount of dilution.Every tank feed supplement needs nutritive salt urea 110g and monoammonium phosphate 20g, and two kinds of nutritive salt are made into 500mL solution together, adds 50% of the total liquid measure of fermentation time 0h～8h average flow Ensure Liquid salt, 50% of the total liquid measure of 8h～12h average flow Ensure Liquid salt by stream.Wherein, the dilution molasses that stream adds and nutritive salt all should 121 DEG C of high-temperature sterilization 20min.
Step 4: by fermentation liquid through whizzer 5000r/min, centrifuge washing 10min, the centrifuge washing 3～5 times of adding water, obtains the yeast extract fresh yeast emulsifiable paste of solid content 60%～70%, then this emulsifiable paste is diluted with water to 10～15% emulsion.
Result: fermentation culture finishes, the barm cell concentration in fermentation liquid reaches 295.54g/L, and thalline size evenly, gemma rate 0.06%, bacterium protein content 55.2%.
example 4
The method of fermentative production high-density yeast extract emulsion, is made up of following steps:
Step 1: bread yeast slant strains is inoculated in 100mL molasses liquid seed culture medium, at 28 DEG C of temperature, intermittently sways and cultivate 24h.
Step 2: the seed liquor 100mL of step 1 is seeded in the 3L triangular flask that molasses liquid seed culture medium is housed to preparation 1000mL seed liquor by 10% inoculum size.At 28 DEG C of temperature, intermittently sway and cultivate 24h.
Wherein, the molasses liquid seed culture medium formula in step 1 and step 2 is: dilution molasses 150mL/L, urea 2.50g/L, monoammonium phosphate 0.60g/L, magnesium sulfate heptahydrate 0.12g/L, Zinc Sulphate Heptahydrate 0.05g/L, Repone K 0.10g/L, sodium-chlor 0.10g/L, micro-mother liquor 2mL.Regulate medium pH to 4.2,121 DEG C of high-temperature sterilization 20min.
Step 3: the seed liquor of step 2 to being equipped with in the 15L fermentor tank of fermention medium, is made to the about 6200mL of initial incubation liquid cumulative volume by inoculum size aseptic inoculation under flame ring of 15%.Regulate 28 DEG C of temperature, pH3.5～4.5,0.1～0.3vvm is aerlbic culture 6h in a small amount.Start afterwards flow feeding dilution molasses and nutritive salt.Feeding culture condition: culture temperature is 28 DEG C, pH3.5～4.5, under air flow 1.0～2.0vvm, feed supplement stream adds 24h.
Wherein, the fermentative medium formula of 15L fermentor tank is: dilution molasses 200mL, urea 10.0g, monoammonium phosphate 9.0g, magnesium sulfate heptahydrate 7.5g, Zinc Sulphate Heptahydrate 1.0g, Repone K 5g, sodium-chlor 5g, micro-mother liquor 10mL.121 DEG C of high-temperature sterilization 20min.
Trace element mother liquor formula: MnCI
22H
2o 4mg/L, KI 0.4 mg/L, pantothenic acid 4mg/L, vitamin H 0.1 mg/L, vitamins B
10.4 g/L, vitamins C 4 mg/L.
Feeding medium during fermentation formula and feeding method control: every tank stream adds the about 5.0L of dilution molasses, add fermentation time 0h～8h average flow by stream and add 20% of the sweet total amount of dilution, and 8h～18h average flow adds 56% of the sweet total amount of dilution, and 18h～25h average flow adds 24% of the sweet total amount of dilution.Every tank feed supplement needs nutritive salt urea 110g and monoammonium phosphate 20g, and two kinds of nutritive salt are made into 500mL solution together, adds 50% of the total liquid measure of fermentation time 0h～8h average flow Ensure Liquid salt, 50% of the total liquid measure of 8h～12h average flow Ensure Liquid salt by stream.Wherein, the dilution molasses that stream adds and nutritive salt all should 121 DEG C of high-temperature sterilization 20min.
Step 4: by fermentation liquid through whizzer 5000r/min, centrifuge washing 10min, the centrifuge washing 3～5 times of adding water, obtains the yeast extract fresh yeast emulsifiable paste of solid content 60%～70%, then this emulsifiable paste is diluted with water to 10～15% emulsion.
Result: fermentation culture finishes, the barm cell concentration in fermentation liquid reaches 300.08g/L, and thalline size is evenly, gemma rate 1.1%, bacterium protein content 55.0%, makes seasonings raciness.
Claims (8)
1. a method for fermentative production high-density yeast extract emulsion, is characterized in that, comprises the steps:
A). bread yeast slant strains is inoculated in 100mL molasses liquid seed culture medium, at 28 DEG C of temperature, intermittently sways and cultivate 24h or slow speed of revolution 100rpm shaking culture 24h;
B). the seed liquor 100mL of step a) is seeded in the 3L triangular flask that molasses liquid seed culture medium is housed to preparation 1000mL seed liquor by 10% inoculum size.
2. at 28 DEG C of temperature, intermittently sway and cultivate 24h or slow speed of revolution 100rpm shaking culture 24h;
C). the seed liquor of step b) to being equipped with in the 15L fermentor tank of fermention medium, is made to the about 6200mL of initial incubation liquid cumulative volume by inoculum size aseptic inoculation under flame ring of 15%; Regulate 28 DEG C of temperature, pH3.5～4.5,0.1～0.3vvm is aerlbic culture 4h～6h in a small amount; Start afterwards by processing requirement flow feeding dilution molasses and nutritive salt, feeding culture condition: culture temperature is 28 DEG C, pH3.5～4.5, under air flow 1.0～2.0vvm, feed supplement stream adds 24h～25h;
D). the fermentation liquid that step c) is obtained is through whizzer 5000r/min, centrifuge washing 10min, add water centrifuge washing 3～5 times, obtains the yeast extract fresh yeast emulsifiable paste of solid content 60%～70%, then this fresh yeast emulsifiable paste is diluted with water to 10～15% emulsion;
Method according to claim 1, it is characterized in that, the molasses liquid seed culture medium formula in step a) and step b) is: dilution molasses 150mL/L, urea 2.50g/L, monoammonium phosphate 0.60g/L, magnesium sulfate heptahydrate 0.12g/L, Zinc Sulphate Heptahydrate 0.05g/L, Repone K 0.10g/L, sodium-chlor 0.10g/L, trace element mother liquor 2mL, regulates medium pH to 4.2,121 DEG C of high-temperature sterilization 20min.
3. method according to claim 1, it is characterized in that, fermentative medium formula in step c) is: dilution molasses 200mL, urea 10.0g, monoammonium phosphate 9.0g, magnesium sulfate heptahydrate 7.5g, Zinc Sulphate Heptahydrate 1.0g, Repone K 5g, sodium-chlor 5g, trace element mother liquor 10mL, 121 DEG C of high-temperature sterilization 20min.
4. according to the method described in claim 1～3 any one, it is characterized in that, described dilution molasses are that former molasses are diluted with water to hammer degree 45～50%oBx, heated and boiled 1min, is cooled to 50～80 DEG C, uses whizzer 4000r/min, centrifugation 10min, discards throw out, obtains dilution molasses.
5. method according to claim 1, is characterized in that, the stream Ensure Liquid salt in step c) is, urea 110g and monoammonium phosphate 20g, and two kinds of nutritive salt are made into 500mL solution together, 121 DEG C of high-temperature sterilization 20min.
6. method according to claim 1, is characterized in that, it is through 121 DEG C of high-temperature sterilization 20min that the stream in step c) adds dilution molasses.
7. according to the method described in claim 2～3 any one, it is characterized in that, described micro-mother liquor formula is: MnCI
22H
2o 4mg/L, KI 0.4 mg/L, pantothenic acid 4mg/L, vitamin H 0.1 mg/L, vitamins B
10.4 g/L, vitamins C 4 mg/L.
8. method according to claim 1, it is characterized in that, described in step c) by processing requirement flow feeding dilution molasses and nutritive salt: every tank stream adds the about 5.0L of dilution molasses, add fermentation time 0h～8h average flow by stream and add 20% of the sweet total amount of dilution, 8h～18h average flow adds 56% of the sweet total amount of dilution, and 18h～24h average flow adds 24% of the sweet total amount of dilution; Every tank feed supplement needs nutritive salt urea 110g and monoammonium phosphate 20g, and two kinds of nutritive salt are made into 500mL solution together, adds 50% of the total liquid measure of fermentation time 0h～8h average flow Ensure Liquid salt, 50% of the total liquid measure of 8h～12h average flow Ensure Liquid salt by stream.
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