CN102643878A - Method for producing gamma-polyglutamic acid by utilizing fermentation of bacillus natto - Google Patents
Method for producing gamma-polyglutamic acid by utilizing fermentation of bacillus natto Download PDFInfo
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- CN102643878A CN102643878A CN2012101350732A CN201210135073A CN102643878A CN 102643878 A CN102643878 A CN 102643878A CN 2012101350732 A CN2012101350732 A CN 2012101350732A CN 201210135073 A CN201210135073 A CN 201210135073A CN 102643878 A CN102643878 A CN 102643878A
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Abstract
The invention provides a method for producing gamma-polyglutamic acid by utilizing fermentation of bacillus natto. The optical fermentation culture condition of strains is determined through research for culture conditions of strains. The optical fermentation culture condition includes 25g/L of sucrose, 5g/L of yeast extract, 40g/L of monosodium glutamate, 1g/L of Na2HPO4, 1g/L of NaH2PO4, 0.5g/L of MgSO4, an Erlenmeyer flask with loaded fluid of 70mL/250mL, and initial pH 6.5, adding 4% of NaCl to the strain at 37 DEG C and 120 r/min for 24 hours, and continuous culture for 24 hours enables the yield of the gamma-polyglutamic acid to reach 18.04g/L.
Description
Technical field
The invention belongs to fermentation technical field, be specifically related to a kind of method of utilizing bacillus natto to ferment to produce gamma-polyglutamic acid-.
Background technology
Gamma-polyglutamic acid-is to pass through a kind of water miscible bioabsorbable polymer material that γ-amido linkage combination forms by L-L-glutamic acid or D-L-glutamic acid.Because gamma-polyglutamic acid-has water-absorbent, degradability, numerous characteristics such as water-disintegrable, therefore is widely used in multiple fields such as medicine, food-processing, agricultural, greening and water treatment, has great exploitation value and application prospect.
The initial discovery of gamma-polyglutamic acid-is to exist as a kind of staple of Bacillus anthracis pod membrane, and people found successively that multiple genus bacillus can both produce gamma-polyglutamic acid-outside born of the same parents afterwards.Because easy cultivation property and the security of bacillus natto, Bacillus licheniformis, the production of polyglutamic acid now report is mostly produced by this type bacterial strain.Although many investigators have carried out big quantity research to the Production by Microorganism Fermentation gamma-polyglutamic acid-, productive rate generally also is in lower level, and production cost is high, this to a great extent limit the industrial production and the application of gamma-polyglutamic acid-.Because the biosynthetic mechanism of gamma-polyglutamic acid-is still not really clear so far, and the pathways metabolism of bacterial strain generation gamma-polyglutamic acid-is complicated, and regulative mode is various, and the method that improves its production efficiency at present mainly relies on seed selection excellent species and optimization for fermentation technology.
Fermentation is a kind of biological process of complicacy, receives the influence of several factors, all with the output of purpose product confidential relation is arranged like quality, fermentation condition and the fermenting process control etc. of the proportioning of the production performance of thalline self, substratum, seed.In industrial production,, optimum production substratum and culture condition should be provided in order to bring into play the throughput of thalline to greatest extent.Therefore, explore basis that optimal culture conditions that the bacterial strain high yield characteristics gives full expression to is a fermentative prodn and crucial.
Summary of the invention
The object of the present invention is to provide a kind of method of utilizing bacillus natto to ferment to produce gamma-polyglutamic acid-; Through to the strain culturing Study on Conditions, confirmed the righttest fermentation culture conditions of bacterial strain: sucrose 25g/L, yeast extract paste 5g/L; Sodium Glutamate 40g/L, Na
2HPO
41g/L, NaH
2PO
41g/L, MgSO
40.5g/L, liquid amount 70mL/250 mL Erlenmeyer flask, initial pH is 6.5, and bacterial classification adds 4% NaCl at 37 ℃, 120r/min after cultivating 24h, continues to cultivate 24 h again, and the output of gamma-polyglutamic acid-can reach 18.04 g/L.
A kind of method of utilizing bacillus natto to ferment to produce gamma-polyglutamic acid-; It is characterized in that seed liquor is inserted in 3% (v/v) ratio in the Erlenmeyer flask of the 250mL that the 50mL fermention medium is housed, 37 ℃, 120r/min are behind the cultivation 24h; In substratum, add 4%NaCl, continue to cultivate 24h.
1. the described method of step 1, concrete steps are following:
(1) picking one collarium is inoculated into the 250mL Erlenmeyer flask that the 50mL seed culture medium is housed from bacterial strain refrigeration inclined-plane with transfering loop, and 37 ℃, 120r/min cultivates 24h.
(2) the 250mL Erlenmeyer flask 50mL fermention medium of packing into inserts seed liquor by 2% inoculum size, and bacterial classification is at 37 ℃, 120r/min, cultivate 24h after, in substratum, add NaCl, continue to cultivate 24h, measure gamma-polyglutamic acid-output.
3. the said seed culture medium of step 2 (g/L): peptone 10, Carnis Bovis seu Bubali cream 5, NaCl 10, zero(ppm) water 1000mL.
4. the described fermention medium of step 2 (g/L): sucrose 25, yeast extract paste 5, Sodium Glutamate 40, Na
2HPO
41, NaH
2 PO
41, MgSO
40.5, zero(ppm) water 1000mL.
5. the described ph optimum of step 2 (2) is 6.5.
6. the described NaCl addition of step 2 (2) is 4%.
7. the described optimum inoculum size of step 2 (2) is 3%.
Description of drawings
Relation between Fig. 1 different vaccination amount and the gamma-polyglutamic acid-output.
Relation between different original ph of Fig. 2 and the gamma-polyglutamic acid-output.
Embodiment
Following embodiment elaborates to the present invention, but to not restriction of the present invention.
The mikrobe bacillus natto (food grade) that uses of the present invention is bought farming hat (joint) bio tech ltd from Guangzhou.
Present embodiment explanation different vaccination amount is produced gamma-polyglutamic acid-to fermentation influence; Seed liquor transferred with 1%, 2%, 3%, 4%, 5% inoculum size carry out fermentation culture in the 50mL fermention medium; After cultivating 24h, in substratum, add 4%NaCl, continue to cultivate 24h.
Can find out by Fig. 1; The gamma-polyglutamic acid-output that when inoculum size is 3%, obtains is the highest; When inoculum size is respectively 1%, 2%, 3%; The output of the final gamma-polyglutamic acid-in the fermented liquid is explained and has been strengthened inoculum size along with the increase of inoculum size increases, and helps the quick growth of gamma-polyglutamic acid-; Inoculum size was at 4%, 5% o'clock, and the gamma-polyglutamic acid-output in the fermented liquid descends on the contrary, possibly be the increase along with inoculum size; Metabolic waste by bringing in the seed culture medium increases; And thalli growth is too fast, and the cell decline is quickened, thereby influences gamma-polyglutamic acid-output.
The influence that gamma-polyglutamic acid-is produced in fermentation to thalline of present embodiment explanation different fermentations liquid original ph; The original ph of regulating fermention medium is respectively: 4.0,4.5,5.0,5.5,6.0,6.5,7.0; After cultivating 24h; In substratum, add 4%NaCl, continue to measure fermented liquid gamma-polyglutamic acid-content behind the cultivation 24h.
The optimum pH of microorganism growth stage and product synthesis phase is normally different, and this is not only relevant with strain properties, and is also relevant with the chemical property of product.Can be known by Fig. 2, be 6.5 o'clock in original ph, and the output of gamma-polyglutamic acid-is higher relatively, and therefore, this paper is decided to be 6.5 with the initial optimum pH of fermention medium.
The influence that the different additions of present embodiment explanation NaCl produce gamma-polyglutamic acid-to fermentation in the above under other optimal conditionss, is added 0%, 2%, 4%, 6%, 8% NaCl respectively and is carried out fermentation culture in fermention medium, measure gamma-polyglutamic acid-output.
Visible by table 1, along with the increasing of NaCl concentration, the output of gamma-polyglutamic acid-rises gradually, surpasses at 4% o'clock and begins to descend, and can think that gamma-polyglutamic acid-output is the highest when NaCl concentration reaches 4%.
Table 1 NaCl concentration is to the influence of gamma-polyglutamic acid-fermentation
| NaCl concentration (%) | 0% | 2% | 4% | 6% | 8% |
| Gamma-polyglutamic acid-(g/L) | 15.2 | 17.1 | 18.8 | 16.4 | 15.4 |
Claims (7)
1. method of utilizing bacillus natto to ferment to produce gamma-polyglutamic acid-; It is characterized in that inserting seed liquor in the Erlenmeyer flask of the 250mL that the 50mL fermention medium is housed in 3% (v/v) ratio; 37 ℃, 120r/min; After cultivating 24h, in substratum, add 4%NaCl, continue to cultivate 24h.
2. method according to claim 1 is characterized in that concrete steps are following:
(1) picking one collarium is inoculated into the 250mL Erlenmeyer flask that the 50mL seed culture medium is housed from bacterial strain refrigeration inclined-plane with transfering loop, and 37 ℃, 120r/min cultivates 24h;
(2) the 250mL Erlenmeyer flask 50mL fermention medium of packing into inserts seed liquor by 3% inoculum size, and bacterial classification is at 37 ℃, 120r/min, cultivate 24h after, in substratum, add NaCl, continue to cultivate 24h, measure gamma-polyglutamic acid-output.
3. method according to claim 2 is characterized in that seed culture medium (g/L): peptone 10, and Carnis Bovis seu Bubali cream 5, NaCl 10, zero(ppm) water 1000mL.
4. method according to claim 2 is characterized in that fermention medium (g/L): sucrose 25, yeast extract paste 5, Sodium Glutamate 40, Na
2HPO
41, NaH
2PO
41, MgSO
40.5, zero(ppm) water 1000mL.
5. method according to claim 2, the pH that it is characterized in that step (2) is 6.5.
6. method according to claim 2, the NaCl addition that it is characterized in that step (2) is 4%.
7. method according to claim 2, the inoculum size that it is characterized in that step (2) is 3%.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106834190A (en) * | 2017-03-11 | 2017-06-13 | 鲁东大学 | Culture medium, bacterial strain and the method for polyglutamic acid are produced with pea protein wastewater fermentation |
-
2012
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Non-Patent Citations (6)
| Title |
|---|
| 《化学与生物工程》 20071231 李大力等 盐浓度对纳豆芽孢杆菌发酵产gamma-聚谷氨酸影响的研究 50-51、72 1-7 第24卷, 第2期 * |
| 《现代食品科技》 20091231 刘常金等 纳豆芽孢杆菌液体发酵生产gamma-聚谷氨酸 935-939 1-7 第25卷, 第8期 * |
| 《食品科技》 20081231 李艳华等 响应面法优化gamma-聚谷氨酸发酵培养基的研究 45-48 1-7 , 第3期 * |
| 刘常金等: "纳豆芽孢杆菌液体发酵生产γ-聚谷氨酸", 《现代食品科技》 * |
| 李大力等: "盐浓度对纳豆芽孢杆菌发酵产γ-聚谷氨酸影响的研究", 《化学与生物工程》 * |
| 李艳华等: "响应面法优化γ-聚谷氨酸发酵培养基的研究", 《食品科技》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106834190A (en) * | 2017-03-11 | 2017-06-13 | 鲁东大学 | Culture medium, bacterial strain and the method for polyglutamic acid are produced with pea protein wastewater fermentation |
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Application publication date: 20120822 |