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High pH value tolerant bacterial strain and novel fermentation method for producing lysine

A technology of lysine and pH value, which is applied in the field of high pH value-resistant strains and the production of lysine by a new fermentation method, can solve the problems of low lysine yield and low bacterial cells, and achieve high yield, inorganic nitrogen and other problems. Sufficient source to ensure the effect of normal metabolism

Active Publication Date: 2014-08-20
SHOUGUANG GOLDEN FAR EAST MODIFIED STARCH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The pH of the traditional lysine fermentation process is generally controlled to be less than 7.2, not lower than 6.5, and the preferred pH is 7.0. At this time, the production of lysine and the growth of bacteria are normal; too high or too low pH will affect or inhibit the fermentation process The production of lysine and the growth of the bacteria make the bacteria in the end of the fermentation process low, and the yield of lysine in the fermentation is low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The strain uses Corynebacterium glutamicum to carry out continuous fermentation of lysine to produce lysine, and the medium used is as follows:

[0039] (1) Inclined culture medium:

[0040] Peptone 12 g / L, yeast powder 7 g / L, sodium chloride 5 g / L, sucrose 20 g / L, agar 20 g / L, medium with 30% potassium hydroxide solution to adjust pH to 7.5-8.0, at 115 Sterilize at ℃ for 20 minutes to prepare a high pH slant medium.

[0041] (2) Seed shake flask medium:

[0042] Peptone 10 g / L

[0043] Yeast powder 5 g / L

[0044] Sodium chloride 5 g / L

[0045] Glucose 40g / L

[0046] Ammonium sulfate 5g / L

[0047] Urea 3g / L

[0048] Potassium dihydrogen phosphate 3g / L

[0049] FeSO 4 0.1g / L

[0050] MnSO 4 0.1g / L

[0051] Vitamin B 1 10mg / L

[0052] Vitamin H 10mg / L

[0053] Vitamin B 3 20mg / L

[0054] Adjust the pH to 7.5-8.0 with 30% potassium hydroxide solution, and sterilize the medium at 120°C for 15 minutes.

[0055] (3) Medium for producing...

Embodiment 2

[0070] Streak the strain into the slant medium and culture it at 31.5°C for 24-32hr. The strains on the plates were collected and expanded for culture, and the strains with better growth were screened out as the experimental strains. Scrape 1 / 4-1 ring seeds from the well-grown activated slant, insert them into 500ml liquid medium, and put them into a shaker for cultivation. Cultivate for 18-24 hours at a temperature of 35° C. and a rotation speed of 180-220 r. Bring the shake flask seeds to logarithmic growth phase. Take the shake flask seed liquid and put it into a 3L automatic fermenter BIO-110 (NBS company in the United States) containing 1L of fermentation medium at an inoculum amount of 10%. The fermenter has automatic control functions such as temperature, pH, and dissolved oxygen.

[0071] During the fermentation process, the fermentation temperature is controlled at 35°C, the tank pressure is controlled at 0.03Mpa, and the dissolved oxygen is controlled at 15% to 25%...

Embodiment 3

[0073] Bacterial classification, seed preparation and fermentation condition are the same as example two. Among them: during the continuous fermentation process, the method of adding dilute hydrochloric acid is used to control the pH, and the initial pH is controlled at 7.0. The method of increasing the pH is used first to reduce the inorganic nitrogen in the early stage, and the pH is raised to 7.3 at the highest. If the consumption of inorganic nitrogen is fast, and the pH cannot meet the demand of ammonia nitrogen at 7.3, then add an appropriate amount of concentrated hydrochloric acid to control it. The feeding of nutrients was started at 3hr, and the feeding ratio of nutrients (nutrients were organic nitrogen source, phosphoric acid, inorganic salts and vitamins) was 1 / 8-1 / 3 of the amount of glucose supplementation.

[0074] This nutritional formula is formulated by dissolving the following ingredients (1L)

[0075] h 3 PO 4 10g

[0076] MgSO 4 10g

[0077] ...

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Abstract

The invention discloses a high pH value tolerant bacterial strain and a novel fermentation method for producing lysine. A fermentation process by the high pH value tolerant bacterial strain does not use ammonium sulfate as an inorganic nitrogen source, and ammonia or liquefied ammonia is used for supplementing the nitrogen source in the fermentation broth to synthesize lysine. The fermentation process for producing lysine does not employ ammonium sulfate, and ammonia or liquefied ammonia is supplemented as the nitrogen source, so that the sulfate ion concentration in the fermentation broth is reduced greatly to facilitate subsequent treatment of fermentation broth; and purification by ion exchange for removing sulfate radical and other ions is not required, so as to simplify the production process of feed grade lysine hydrochloride and reduce the production cost. Supplement of ammonia or liquefied ammonia adjusts the pH value of the fermentation broth to control the inorganic nitrogen content in the fermentation broth, so as to ensure the content of inorganic nitrogen in the culture tank is in a reasonable range; and no ammonium sulfate is added, thereby reducing sulfate radical ions and other impurities in the fermentation broth, and facilitating extraction.

Description

technical field [0001] The invention belongs to a method for producing lysine by a fermentation method, and in particular relates to a strain resistant to high pH value and a method for producing lysine by a novel fermentation method. Background technique [0002] When the content of L-lysine in food or feed is insufficient, the utilization of other amino acids will be limited, so lysine is widely used in food fortification, animal feed additives and medicinal purposes. L-lysine is currently the The main additive of full-price compound feed can improve feed utilization, promote animal growth and improve meat quality. [0003] At present, in the process of producing feed-grade L-lysine hydrochloride, continuous fermentation culture is often used for fermentation, and in continuous fermentation culture, the method of feeding ammonium sulfate is mainly used to provide nitrogen source to ensure the synthesis of lysine. Nitrogen in ammonium sulfate is gradually consumed, while ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P13/08C12R1/15
Inventor 吴泽华李刚冯雨罗明刚
Owner SHOUGUANG GOLDEN FAR EAST MODIFIED STARCH CO LTD
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