CN105316371B - A method of for improving tryptophan fermentation yield - Google Patents
A method of for improving tryptophan fermentation yield Download PDFInfo
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- CN105316371B CN105316371B CN201510702015.7A CN201510702015A CN105316371B CN 105316371 B CN105316371 B CN 105316371B CN 201510702015 A CN201510702015 A CN 201510702015A CN 105316371 B CN105316371 B CN 105316371B
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Abstract
The invention belongs to technical field of microbial fermentation, a kind of more particularly to method for improving tryptophan fermentation yield, the technical solution of use includes the following steps, fermentation culture is prepared first and L phenylalanines are added into fermentation culture, then it sterilizes to fermentation culture, last inoculation fermentation, fermented and cultured is carried out by feed supplement batch, the innovation of the present invention is that L phenylalanines are added into fermentation culture, its optimum addition is 0.2 0.6g/L, using such technical solution, the yield of tryptophan can be significantly improved, increase conversion ratio, by-product is reduced to generate, improve production capacity, to reduce cost.
Description
Technical field
The present invention relates to technical field of microbial fermentation more particularly to a kind of sides for improving tryptophan fermentation yield
Method.
Background technology
Tryptophan (Tryptophan), tryptophane, for white or yellowish crystallization or crystalline powder；Nothing
It is smelly, mildly bitter flavor.Slightly soluble in water, soluble,very slightly, insoluble in chloroform in ethanol, readily soluble in formic acid, in sodium hydroxide test solution
Or it is dissolved in dilute hydrochloric acid.Tryptophan is the precursor substance that auxin biosynthesis is important in plant, and structure is similar to IAA,
The generally existing in higher plant.
The production of L-Trp is mainly manufactured by chemical synthesis and protein Hydrolyze method earliest.With to microbial method
The continuous development of the research of tryptophan is produced, people start with microbial method fermenting and producing tryptophan, have moved towards practicality simultaneously
And it is in leading position.Microbial method can substantially be divided into microbe fermentation method and enzymatic synthesis.It has also appeared in recent years directly
Fermentation method and chemical synthesis, direct fermentation and conversion method are combined the research of production tryptophan.In addition, genetic engineering, enzyme
Immobilization and the technologies such as High Density Cultivation greatly pushed direct fermentation in the industrial application of Microbial Breeding and enzyme
With the process of industrialization of Production by Enzymes tryptophan.
However the shortcomings such as utilize microbial fermentation production tryptophan that but there is by-products be more, conversion ratio is low, therefore,
It is necessary to provide a kind of method for improving tryptophan fermentation yield.
Invention content
The object of the present invention is to provide a kind of the fermentation of tryptophan is improved by adding L-phenylalanine in culture solution
Yield increases conversion ratio, reduces by-product generation, improves production capacity, and the method to reduce cost, is in the following way
It realizes：L-phenylalanine is added into fermentation culture.
Preferably, the present invention provides a kind of method for improving tryptophan fermentation yield, specifically comprises the following steps：
(1) it prepares fermentation culture and adds L-phenylalanine into the fermentation culture；
(2) into the fermentation culture of addition L-phenylalanine, access strain carries out fermented and cultured.
Wherein, the strain is Recombinant organism CICC 10303, the Recombinant organism
CICC 10303 is purchased from Chinese industrial Culture Collection.
Wherein, the inoculum concentration of the strain be 3%-8%, preferably 5%.
Wherein, the fermentation culture with water including following component by preparing to obtain：Glucose 8-11g/L, phosphoric acid
Potassium dihydrogen 2-4g/L, yeast powder 1-2g/L, magnesium sulfate 3-4g/L, citric acid 3-4g/L, ferrous sulfate 0.1-0.4g/L.
As a preferred option, the fermentation culture with water including following component by preparing to obtain：Glucose 8-
11g/L, potassium dihydrogen phosphate 2-4g/L, yeast powder 1-2g/L, magnesium sulfate 3-4g/L, citric acid 3-4g/L, ferrous sulfate 0.1-
0.4g/L, copper sulphate 0.001-0.002g/L, sodium sulfite 0.02-0.05g/L, cobalt chloride 0.002-0.003g/L.
As most preferred scheme, the fermentation culture with water including following component by preparing to obtain：Glucose
8g/L, potassium dihydrogen phosphate 2g/L, yeast powder 1g/L, magnesium sulfate 3g/L, citric acid 3g/L, ferrous sulfate 0.1g/L, copper sulphate
0.001g/L, sodium sulfite 0.02g/L, cobalt chloride 0.002g/L.
Wherein, the fermentation condition of the fermented and cultured is：Pressure：0.03-0.08MPa, pH：6.4-6.7, air quantity：0.8-
1.5vvm, dissolved oxygen 15-25%, period：45-60h.Preferably pressure 0.05MPa, air quantity 1.1vvm；
Wherein, the addition of the L-phenylalanine is 0.2-0.6g/L.After inventor has found addition L-phenylalanine,
Less addition wants high to the yield and conversion ratio of tryptophan, and with the increase of L-phenylalanine additive amount, yield and conversion ratio are in
Gradual increasing trend, when additive amount reaches 0.6g/L, yield and conversion ratio reach maximum；When additive amount reaches 0.9g/L,
Although yield and conversion ratio, which less add, still wants high, its numerical value is below the additive amount of 0.6g/L, therefore L-phenylalanine
Additive amount be 0.2-0.6g/L.
As preferred embodiments of the present invention, a method of further include before fermentation for improving tryptophan fermentation yield
Fermentation culture to adding L-phenylalanine carries out pretreated step, and the pretreated specific method is：Use hydrogen
The pH that sodium oxide molybdena adjusts fermentation culture is 6-7, and sterilize 20-40min under the conditions of 115-125 DEG C, is then cooled to 30-37 DEG C
.
As preferred embodiments of the present invention, in incubation, in order to ensure being smoothed out for fermentation process, added by stream
It is 0.1-0.2g/L that the mode of glucose, which controls the glucose content range in fermentation culture, in a kind of specific embodiment
In, flow the glucose solution that the glucose added is mass fraction 60%-70%.
As the most preferred scheme of the present invention, a method of for improving tryptophan fermentation yield, include the following steps：
(1) it prepares fermentation culture and sterilizes：The fermentation culture with water including following component by preparing to obtain：
Glucose 8g/L, potassium dihydrogen phosphate 2g/L, yeast powder 1g/L, magnesium sulfate 3g/L, citric acid 3g/L, ferrous sulfate 0.1g/L, sulphur
Sour copper 0.001g/L, sodium sulfite 0.02g/L, cobalt chloride 0.002g/L, L-phenylalanine 0.2-0.6g/L, then adjusting pH is
6-7, sterilize 20-40min under the conditions of 115-125 DEG C, is then cooled to 30-37 DEG C；
(2) it ferments：According to 5% inoculum concentration, Recombinant organism is inoculated into the fermentation culture, is pressed
According to pressure：0.03-0.08MPa；pH：6.4-6.7；Air quantity：0.8-1.5vvm；The CMC model 45-60h of dissolved oxygen 15-25%, training
Further stream adds mass fraction to be the glucose solution of 60%-70% during supporting, to maintain the glucose of fermentation culture
Content range is 0.1-0.2g/L.
Specific implementation mode
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..
Embodiment 1
A method of for improving tryptophan fermentation yield, include the following steps：
(1) it prepares fermentation culture and sterilizes：By glucose 8g/L, potassium dihydrogen phosphate 2g/L, yeast powder 1g/L, magnesium sulfate
3g/L, citric acid 3g/L, ferrous sulfate 0.1g/L, copper sulphate 0.001g/L, sodium sulfite 0.02g/L, cobalt chloride 0.002g/L,
L-phenylalanine 0.2g/L is soluble in water；The pH to 6.5 that above-mentioned fermentation culture is adjusted using sodium hydroxide, then at 121 DEG C
Under the conditions of sterilize 30min, after be cooled to 34 DEG C；
(2) inoculated and cultured：Recombinant organism CICC10303 inoculations are entered into fermentation according to 5% inoculum concentration
In culture solution, the pH value of fermentation culture is adjusted in fermentation process by sodium hydroxide, so that it is maintained 6.4-6.7, passes through stream
Add 60% glucose solution glucose content is made to maintain 0.1-0.2g/L, in addition, control system pressure 0.05MPa, wind
1.1vvm, DO15-25% are measured, by the culture of 54h, tryptophane reaches 36.8g/L, and conversion ratio is up to 14%.
Embodiment 2
A method of for improving tryptophan fermentation yield, include the following steps：
(1) it prepares fermentation culture and sterilizes：By glucose 8g/L, potassium dihydrogen phosphate 2g/L, yeast powder 1g/L, magnesium sulfate
3g/L, citric acid 3g/L, ferrous sulfate 0.1g/L, copper sulphate 0.001g/L, sodium sulfite 0.02g/L, cobalt chloride 0.002g/L,
L-phenylalanine 0.4g/L is soluble in water；The pH to 6.5 that above-mentioned fermentation culture is adjusted using sodium hydroxide, then at 121 DEG C
Under the conditions of sterilize 30min, after be cooled to 34 DEG C；
(2) inoculated and cultured：Recombinant organism CICC10303 inoculations are entered into fermentation according to 5% inoculum concentration
In culture solution, the pH value of fermentation culture is adjusted in fermentation process by sodium hydroxide, so that it is maintained 6.4-6.7, passes through stream
Add 65% glucose solution glucose content is made to maintain 0.1-0.2g/L, in addition, control system pressure 0.08MPa, wind
1.5vvm, DO15-25% are measured, by the culture of 50h, tryptophane reaches 40.2g/L, and conversion ratio is up to 14.6%.
Embodiment 3
A method of for improving tryptophan fermentation yield, include the following steps：
(1) it prepares fermentation culture and sterilizes：By glucose 8g/L, potassium dihydrogen phosphate 2g/L, yeast powder 1g/L, magnesium sulfate
3g/L, citric acid 3g/L, ferrous sulfate 0.1g/L, copper sulphate 0.001g/L, sodium sulfite 0.02g/L, cobalt chloride 0.002g/L,
L-phenylalanine 0.6g/L is soluble in water；The pH to 6.5 that above-mentioned fermentation culture is adjusted using sodium hydroxide, then at 121 DEG C
Under the conditions of sterilize 30min, after be cooled to 34 DEG C；
(2) inoculated and cultured：Recombinant organism CICC10303 inoculations are entered into fermentation according to 5% inoculum concentration
In culture solution, the pH value of fermentation culture is adjusted in fermentation process by sodium hydroxide, so that it is maintained 6.4-6.7, passes through stream
Add 70% glucose solution glucose content is made to maintain 0.1-0.2g/L, in addition, control system pressure 0.08MPa, wind
1.2vvm, DO15-25% are measured, by the culture of 48h, tryptophane reaches 46.8g/L, and conversion ratio is up to 16.2%.
1 prior art of comparative example
A kind of fermentation process of tryptophan, includes the following steps：
(1) it prepares fermentation culture and sterilizes：By glucose 8g/L, potassium dihydrogen phosphate 2g/L, yeast powder 1g/L, magnesium sulfate
3g/L, citric acid 3g/L, ferrous sulfate 0.1g/L, copper sulphate 0.001g/L, sodium sulfite 0.02g/L, cobalt chloride 0.002g/L
It is soluble in water；The pH to 6.5 of above-mentioned fermentation culture is adjusted using sodium hydroxide, then sterilize under the conditions of 121 DEG C 30min,
After be cooled to 34 DEG C；
(2) inoculated and cultured：Recombinant organism CICC10303 inoculations are entered into fermentation according to 5% inoculum concentration
In culture solution, the pH value of fermentation culture is adjusted in fermentation process by sodium hydroxide, so that it is maintained 6.4-6.7, passes through stream
Add 65% glucose solution glucose content is made to maintain 0.1-0.2g/L, in addition, control system pressure 0.08MPa, wind
1.5vvm, DO15-25% are measured, by the culture of 54h, tryptophane 35.2g/L, conversion ratio 13.6%.
Comparative example 2 investigates influence of the addition of L-phenylalanine to raising tryptophan yield
A kind of fermentation process of tryptophan, includes the following steps：
(1) it prepares fermentation culture and sterilizes：By glucose 8g/L, potassium dihydrogen phosphate 2g/L, yeast powder 1g/L, magnesium sulfate
3g/L, citric acid 3g/L, ferrous sulfate 0.1g/L, copper sulphate 0.001g/L, sodium sulfite 0.02g/L, cobalt chloride 0.002g/L,
L-phenylalanine 0.9g/L is soluble in water；The pH to 6.5 that above-mentioned fermentation culture is adjusted using sodium hydroxide, then at 121 DEG C
Under the conditions of sterilize 30min, after be cooled to 34 DEG C；
(2) inoculated and cultured：Recombinant organism CICC10303 inoculations are entered into fermentation according to 5% inoculum concentration
In culture solution, the pH value of fermentation culture is adjusted in fermentation process by sodium hydroxide, so that it is maintained 6.4-6.7, passes through stream
Add 65% glucose solution glucose content is made to maintain 0.1-0.2g/L, in addition, control system pressure 0.08MPa, wind
1.5vvm, DO15-25% are measured, by the culture of 48h, tryptophane 38.4g/L, conversion ratio 14.3%.
Although above having used general explanation, specific implementation mode and experiment, the present invention is made to retouch in detail
It states, but on the basis of the present invention, it can be made some modifications or improvements, this is apparent to those skilled in the art
's.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to claimed
Range.
Claims (1)
1. a kind of method for improving tryptophan fermentation yield, it is characterised in that：Include the following steps：
(1) it prepares fermentation culture and sterilizes：The fermentation culture is prepared to obtain by following component with water：Glucose 8g/L,
Potassium dihydrogen phosphate 2g/L, yeast powder 1g/L, magnesium sulfate 3g/L, citric acid 3g/L, ferrous sulfate 0.1g/L, copper sulphate 0.001g/
L, sodium sulfite 0.02g/L, cobalt chloride 0.002g/L, L-phenylalanine 0.2-0.6g/L, it is 6-7 then to adjust pH, in 115-
Sterilize 20-40min under the conditions of 125 DEG C, is then cooled to 30-37 DEG C；
(2) it ferments：According to 5% inoculum concentration, Escherichia coli (Escherichia coli) genetic engineering bacterium CICC10303 is connect
In kind to the fermentation culture, according to pressure：0.03-0.08MPa；pH：6.4-6.7；Air quantity：0.8-1.5vvm；Dissolved oxygen 15-
25% CMC model 45-60h, further stream plus the glucose solution that mass fraction is 60%-70% in incubation, with
Maintain the glucose content ranging from 0.1-0.2g/L of fermentation culture.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107058417B (en) * | 2016-06-27 | 2021-04-06 | 通辽梅花生物科技有限公司 | Novel process for improving quality and enhancing efficiency of tryptophan |
| CN106755159A (en) * | 2016-12-21 | 2017-05-31 | 天津科技大学 | A kind of liquor fermentation culture medium and the method for improving L tryptophan yield |
| CN106967761A (en) * | 2017-05-27 | 2017-07-21 | 新疆苏源生物工程有限公司 | L phenylalanines and preparation method thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1200150A (en) * | 1995-08-30 | 1998-11-25 | 味之素株式会社 | Process for producing L-amino acids |
| CN101622338A (en) * | 2006-12-29 | 2010-01-06 | Cj第一制糖株式会社 | Genetically engineered recombinant escherichia coli producing l-tryptophan having originally l-phenylalanine productivity, and method for producing l-tryptophan using the microorganism |
| CN101838628A (en) * | 2001-11-23 | 2010-09-22 | 味之素株式会社 | Utilize the escherichia bacterium to produce the amino acid whose method of L- |
| CN102168118A (en) * | 2011-01-31 | 2011-08-31 | 安徽丰原发酵技术工程研究有限公司 | Method for increasing fermentation output of tryptophan |
| CN102978252A (en) * | 2011-09-02 | 2013-03-20 | 孟州市华兴生物化工有限责任公司 | L-tryptophan fed-batch fermentation technology |
-
2015
- 2015-10-23 CN CN201510702015.7A patent/CN105316371B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1200150A (en) * | 1995-08-30 | 1998-11-25 | 味之素株式会社 | Process for producing L-amino acids |
| CN101838628A (en) * | 2001-11-23 | 2010-09-22 | 味之素株式会社 | Utilize the escherichia bacterium to produce the amino acid whose method of L- |
| CN101622338A (en) * | 2006-12-29 | 2010-01-06 | Cj第一制糖株式会社 | Genetically engineered recombinant escherichia coli producing l-tryptophan having originally l-phenylalanine productivity, and method for producing l-tryptophan using the microorganism |
| CN102168118A (en) * | 2011-01-31 | 2011-08-31 | 安徽丰原发酵技术工程研究有限公司 | Method for increasing fermentation output of tryptophan |
| CN102978252A (en) * | 2011-09-02 | 2013-03-20 | 孟州市华兴生物化工有限责任公司 | L-tryptophan fed-batch fermentation technology |
Non-Patent Citations (3)
| Title |
|---|
| Development of l-tryptophan production strains by defined genetic modification in Escherichia coli;Zhi-Jun Zhao等;《J Ind Microbiol Biotechnol》;20110504;第38卷(第12期);第1921-1929页 * |
| L-色氨酸生产菌株的构建及代谢调控;赵志军;《中国博士学位论文全文数据库（电子期刊）工程科技I辑》;20120615;第13页"2.2.1菌株和质粒"，第15页"2.2.3.1培养基"，第40页表3-1，第47页"3.3.6工程菌FB-04/pSV04的补料分批发酵" * |
| 不同添加物对L-色氨酸补料分批发酵的影响;杨海军 等;《天津科技大学学报》;20040331;第19卷(第1期);第1-3，8页 * |
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