CN1977048B - Fermentation of L-lysine using acid-treated product of soybean meal - Google Patents
Fermentation of L-lysine using acid-treated product of soybean meal Download PDFInfo
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- CN1977048B CN1977048B CN2005800216202A CN200580021620A CN1977048B CN 1977048 B CN1977048 B CN 1977048B CN 2005800216202 A CN2005800216202 A CN 2005800216202A CN 200580021620 A CN200580021620 A CN 200580021620A CN 1977048 B CN1977048 B CN 1977048B
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- fermentation
- methionin
- dregs
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- 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
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/04—Alpha- or beta- amino acids
- C12P13/08—Lysine; Diaminopimelic acid; Threonine; Valine
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Abstract
Disclosed is a method for fermentation of L-lysine using acid hydrolyzate of soybean meal as organic nitrogen source. The method comprises adjusting pH of the acid hydrolyzate of soybean meal to pH 2~4 before preparing a culture medium. Such adjusted pH values allows the fermentation level and yield per sugar of L-lysine to be increased by at least 8% and at least 7%, respectively, as compared to other pH values.
Description
Technical field
The present invention relates to be used for the fermentation process of L-Methionin, its use corynebacterium glutamicum as the acid hydrolysis products of the microorganism of fermentation and dregs of beans as organic nitrogen source, it is characterized in that when the preparation substratum, the pH value of the acid hydrolysis products of dregs of beans is adjusted to predeterminated level, so that improve the productivity of L-fermenting lysine.
Background technology
L-Methionin is a kind of primary amino acid and is used as cattle food, foodstuff additive and pharmaceutical raw material.Especially, kept domestic animal feed additive up to about 700,000 tons the total market size at L-Methionin in 2004.Therefore, be appreciated that L-Methionin is large-scale biotechnology product.Recently, as L-Methionin every year of domestic animal feed additive with 8～10% increase in demand.In addition, because China seriously goes through increasing rapidly of Ji and the restriction of cattle waste is become more and more stricter, the market of expection L-Methionin can experience expansion rapidly.Therefore, need improve the international competitive ability of L-Methionin product by technical study and exploitation fermentation process and employed therein bacterial strain.
At present, L-Methionin is to utilize carbon source, nitrogenous source and mineral industrial by the direct fermentation method as the raw material of substratum.In the component of substratum, nitrogenous source (for example, yeast extract paste, peptone, corn steep liquor and hydrolyzate of soybean meal) has been concentrated to be used for their kind and the research of processing, so that save cost.In addition, nitrogen sources known has a large amount of spaces and is used for hereinafter improvement.
Simultaneously, dregs of beans is meant from soybean and reclaims after the soybean oil and remaining cheap byproducts.This dregs of beans is with proteolytic enzyme or acid for example behind hydrochloric acid or the sulphuric acid hydrolysis and be used for substratum.There is two types the technology that is used for this acidolysis.
Korean Patent discloses 1996-0017840 number application that has disclosed by the hydrolyzate of soybean meal that obtains with the protease treatment dregs of beans.The method that is used for acid hydrolysis of soybean meal also is known.An example of these methods may further comprise the steps: acid is joined in the dregs of beans, make this mixture react 10～24h under 110～120 ℃ high temperature; Reaction mixture; And provide synthetic product as the culture medium raw material after leaching solid.
The method that above-mentioned use enzyme is used for the dregs of beans hydrolysis is metastable method.Yet it is limited in significantly and is used for the industrial practical application of L-Methionin.For example, most of industrial fermentation factory uses the continuous sterilization device, is used for the substratum sterilization, and needs a plurality of pipelines that are used to finish basic step.Yet, obtaining hydrolyzate of soybean meal by the enzyme that contains multiple peptide or non-hydrolyzed protein solids, squamous (scale) deposition can take place in it, thereby causes serious problem.
Simultaneously, though because the processing of sulfuric acid or hydrochloric acid, above-mentioned acid hydrolysis method needs acid-resistant system, and it has guaranteed the technology stability that brings owing to the dregs of beans complete hydrolysis.Therefore, a part of amino acid Products Co., Ltd comprises Ajinomoto Co., uses the hydrolysate of this dregs of beans.
Summary of the invention
Technical problem
Therefore, made the present invention to solve the problems referred to above that occur in the prior art.An object of the present invention is to provide a kind of method that is used to prepare L-Methionin, it produces the acid hydrolysis products of L-Methionin and dregs of beans by corynebacterium glutamicum or dust Xi Shi intestinal bacteria (E.coli), so that obtain higher fermentation productivity.
After the advantage of having studied the dregs of beans acid hydrolysis products, the applicant finds when the preparation substratum, being adjusted to predeterminated level by pH value and can improving fermentation productivity, make microorganism more effectively utilize hydrolysate, other nutrient and the mineral of dregs of beans the preformed acid hydrolysis products of dregs of beans. this is a new solution of not knowing as yet in the prior art.
Technical scheme
According to an aspect of the present invention, provide a kind of acid hydrolysis products to improve the method for the fermentation productivity of L-Methionin by corynebacterium glutamicum or dust Xi Shi intestinal bacteria generation L-Methionin and dregs of beans.According to another aspect of the present invention, provide the cultured products that a kind of method by above-mentioned increase fermentation productivity obtains and the method for producing L-Methionin.
Hereinafter, will be by explaining the features and advantages of the present invention in further detail with reference to appended examples.
Beneficial effect
As mentioned above, the present invention relates to a kind of acid hydrolysis products that utilizes dregs of beans is used for the L-fermenting lysine as organic nitrogen source method.According to the present invention, when preparation during substratum, compare with other pH value, it is 2～4 that the pH value of hydrolyzate of soybean meal is adjusted to the pH value, can be respectively with fermentation level and every sugar yield increase at least 8% and at least 7% of L-Methionin.
Description of drawings
Fig. 1 exemplary illustration be used to prepare the flow process of the acid hydrolysis products of dregs of beans; And
Fig. 2 illustrates the influence figure of the pH value of hydrolyzate of soybean meal to the L-fermenting lysine.
Embodiment
Embodiment 1: the preparation of the acid hydrolysis products of dregs of beans
Under 40～50 ℃ by with 8% or more total nitrogen content and 50% or the defatted soybean meal of more non-purifying protein content be dissolved in the solution of soybean meal for preparing 10～20% (w/v) concentration in the process water as solvent.20～30% vitriolization solution of soybean meal with being adjusted to ultimate density makes the further thermal treatment 5～10min of mixture by being heated 110～120 ℃ then.Then, will be cooled to 50～60 ℃ through vitriolization and heat treated solution of soybean meal, and with having 150～200 purpose strainer filterings.With the raw material (referring to Fig. 1) of gained filtrate as employed substratum in following fermentation test.
Here, the concentration of L-Methionin is measured by HPLC (high performance liquid chromatography), and the cell quantity in substratum is by 3, centrifugation 15min under the 000rpm and being determined.In case of necessity, by the quantitative certainty equivalence number of Bertrand method.
Embodiment 2: according to the pH value of the acid hydrolysis products of the dregs of beans fermentation productivity of L-Methionin relatively
The acid hydrolysis products of the dregs of beans that is obtained by embodiment 1 carries out seed culture (flask) with the microorganism of product L-Methionin in the shaking culture case, it is by the bacterial strain of applicant with the sequence number preservation of KFCC11043, and carries out seed culture in the small fermentor of 5L.Then, in the 30L fermentor tank, carry out fermentation test.Finish following test in the fed-batch fermentation mode, wherein add extra sugar and additive in the training period.
1) is used for the substratum of seed culture: composition and condition (flask)
With sugared 50g/L, peptone 10g/L, yeast extract paste 10g/L, KH
2PO
41.5g/L, MgSO
47H
2O 0.5g/L, urea 5g/L and vitamin H 50 μ g/L (before sterilization, the pH value is 7.0) cultivated 20～25 hours under 30～32 ℃, 220rpm.
2) be used for the substratum of seed culture: composition and condition (5L fermentor tank)
With sugared 70g/L, yeast extract paste 5g/L, (NH
4)
2SO
410g/L, KH
2PO
41.0g/L, MgSO
47H
2O 0.5g/L, vitamin H 300 μ g/L, thiamines HCl 1,000 μ g/L, niacinamide 2,000 μ g/L, pantothenic acid 500 μ g/L and defoamer 3ml/L are 7.0～7.3 times (regulating by gaseous ammonia) with the ventilation speed cultivation of the stirring velocity of 450～600rpm and 0.5～1vvm (volume of air/culture volume/minute) 20～25 hours at 30～32 ℃, pH value.
3) be used for the substratum of fermentation test: composition and condition (30L fermentor tank)
Acid hydrolysis products 30g/L (becoming different pH values), (NH with sugared 165g/L, molasses (as reducing sugar) 165g/L, dregs of beans with the NaOH formulations prepared from solutions
4)
2SO
450g/L, KH
2PO
42.0g/L, MgSO
47H
2O 1.5g/L, FeSO
47H
2O 50mg/L, MnSO
45H
2O 50mg/L, vitamin H 300 μ g/L, thiamines HCl 1,000 μ g/L, niacinamide 2,000 μ g/L, pantothenic acid 500 μ g/L and defoamer 3m1/L are 7.0～7.3 times (regulating by gaseous ammonia) with the stirring velocity of 450～600rpm and the ventilation speed cultivation of 0.5～1vvm at 30～33 ℃, pH value.
With the acid hydrolysis products of the different pH value preparation dregs of beans shown in the following table 1, be used as the raw material of the substratum under the condition of describing then in the above.
The result, with traditional method the pH value is adjusted to 7.0 or other value under the test carried out compare, when the pH value with the hydrolysate of dregs of beans was adjusted to 2～4, it can make fermentation level and every sugar yield increase at least 8% and at least 7% (referring to Fig. 2) of L-Methionin respectively.
Table 1
The pH value of the acid hydrolysis products of dregs of beans is to the influence of fermentation
Industrial applicibility
As mentioned above, the present invention is very useful for food industry, compare with other pH value since with the pH value of the acid hydrolysis products of dregs of beans be adjusted to 2～4 so that its can be respectively with fermentation level and every sugar yield increase at least 8% and at least 7% of 1B.
Claims (2)
1. a fermentation process that utilizes the acid hydrolysis products of dregs of beans as the L-Methionin of organic nitrogen source is characterized in that, when the microorganism strains KFCC 11043 preparation substratum that utilize to produce L-Methionin, the pH value of hydrolyzate of soybean meal is adjusted to 2～4.
2. method that is used to produce L-Methionin, described L-Methionin is the cultured products that is obtained by method according to claim 1.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020040051945A KR100614219B1 (en) | 2004-07-05 | 2004-07-05 | Fermentation of L-lysine using acid-treated product of soybean meal |
| KR1020040051945 | 2004-07-05 | ||
| KR10-2004-0051945 | 2004-07-05 | ||
| PCT/KR2005/002140 WO2006004371A1 (en) | 2004-07-05 | 2005-07-05 | Fermentation of l-lysine using acid-treated product of soybean meal |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1977048A CN1977048A (en) | 2007-06-06 |
| CN1977048B true CN1977048B (en) | 2010-05-12 |
Family
ID=35783133
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2005800216202A Active CN1977048B (en) | 2004-07-05 | 2005-07-05 | Fermentation of L-lysine using acid-treated product of soybean meal |
Country Status (4)
| Country | Link |
|---|---|
| KR (1) | KR100614219B1 (en) |
| CN (1) | CN1977048B (en) |
| BR (1) | BRPI0512761B1 (en) |
| WO (1) | WO2006004371A1 (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103436566A (en) * | 2013-07-31 | 2013-12-11 | 长治市金泽生物工程有限公司 | Bean pulp hydrolysis technology |
| KR101691435B1 (en) | 2015-03-30 | 2016-12-30 | 배재천 | Kintting yarn having retroreflectivity and retroreflective knitted fabric using the same |
| CN110499261A (en) * | 2018-05-17 | 2019-11-26 | 卢松 | The preparation process of protein enzymatic hydrolyzate and its application in fermented and cultured |
| CN111485009B (en) * | 2020-04-10 | 2022-03-18 | 上海农乐生物制品股份有限公司 | Preparation method of fermentation medium of shenqinmycin and fermentation medium |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5179010A (en) * | 1985-06-10 | 1993-01-12 | Ajinomoto Co., Inc. | Fermentation process for producing L-lysine |
| US5302521A (en) * | 1991-10-21 | 1994-04-12 | Kyowa Hakko Kogyo Co., Ltd. | Process for producing L-lysine by iodothyronine resistant strains of Mucorynebacterium glutamicum |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR810001746B1 (en) * | 1977-07-08 | 1981-11-11 | 다까다 히로시 | Process for the production of l-lysine by fermentation |
| KR100273950B1 (en) * | 1998-09-22 | 2001-01-15 | 손경식 | Corynebacterium glutamicum CJ31-0210 producing L-lysine and method for producing L-lysine using the same |
-
2004
- 2004-07-05 KR KR1020040051945A patent/KR100614219B1/en active IP Right Grant
-
2005
- 2005-07-05 BR BRPI0512761-0A patent/BRPI0512761B1/en active IP Right Grant
- 2005-07-05 WO PCT/KR2005/002140 patent/WO2006004371A1/en active Application Filing
- 2005-07-05 CN CN2005800216202A patent/CN1977048B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5179010A (en) * | 1985-06-10 | 1993-01-12 | Ajinomoto Co., Inc. | Fermentation process for producing L-lysine |
| US5302521A (en) * | 1991-10-21 | 1994-04-12 | Kyowa Hakko Kogyo Co., Ltd. | Process for producing L-lysine by iodothyronine resistant strains of Mucorynebacterium glutamicum |
Non-Patent Citations (2)
| Title |
|---|
| EKWEALOR I.A..Preliminary study of L-lysine production by Bacillus speciesusing various agricultrual by-products.Nahrung47 4.2003,47(4),全文. |
| EKWEALOR I.A..Preliminary study of L-lysine production by Bacillus speciesusing various agricultrual by-products.Nahrung47 4.2003,47(4),全文. * |
Also Published As
| Publication number | Publication date |
|---|---|
| BRPI0512761A (en) | 2008-04-08 |
| CN1977048A (en) | 2007-06-06 |
| KR100614219B1 (en) | 2006-08-21 |
| BRPI0512761B1 (en) | 2018-04-03 |
| KR20060003148A (en) | 2006-01-10 |
| WO2006004371A1 (en) | 2006-01-12 |
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