Three-level fermentation of lysine and coating products thereof

A lysine and product technology, applied in the field of amino acid fermentation, can solve the problems of increasing the workload of animal feeding, affecting the efficiency of animal absorption and utilization, etc., and achieves the effects of no potential safety hazards, simplified operation, and reduced release speed.

Active Publication Date: 2012-01-18
NINGXIA EPPEN BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when lysine is used as feed, because it is a small molecule nutrient, it can be released into the animal body soon, especially when it is ingested in large quantities, which affects the efficiency of animal absorption and utilization
Usually the solution is to feed the animals small and frequent, but this will increase the workload of animal husbandry

Method used

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  • Three-level fermentation of lysine and coating products thereof
  • Three-level fermentation of lysine and coating products thereof
  • Three-level fermentation of lysine and coating products thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1 2

[0034] Embodiment 1 Preparation of dihydrodipicolinate synthase variant gene

[0035] According to the sequence we designed, we commissioned Shanghai Sangon Biotechnology Co., Ltd. to synthesize the variant gene encoding dihydrodipicolinate synthase through commercial channels and constructed it into the Escherichia coli-Corynebacterium shuttle plasmid pMS2 (available from the American Type Microorganisms Collection (ATCC), item number ATCC 67189). The cloning process was carried out according to the "Molecular Cloning Experiment Guide" and the operation guide of the commercial reagents used. The brief process is as follows:

[0036] Synthesize nucleic acid fragments of dihydrodipicolinate synthase variant genes by an automatic DNA synthesizer, phosphorylate the 5' ends of these nucleic acid fragments with T4 polynucleotide kinase (purchased from TaKaRa Company), and then equimolar ratio The five nucleic acid fragments were mixed and denatured at 65°C for 5 minutes, annealed ...

Embodiment 2

[0038] The fermentation experiment of embodiment 2 coryneform bacteria

[0039] The pMS2-dap plasmid is transferred into L-lysine-fermenting coryneform bacteria engineering bacteria (available from the American Type Microorganism Collection (ATCC), product number ATCC 31269) by electroporation method, and its brief process is: the coryneform bacteria Shake culture in 50 mL LB liquid medium until OD500 reaches 0.7, collect the cells by centrifugation, wash with 0°C pre-cooled 10% (V / V) glycerol solution, and resuspend the cells in 200 μL pre-cooled 10% (V / V) glycerol solution. V) In glycerol solution, add pMS2-dap plasmid, mix well, transfer to 0.1cm electric shock cup, conduct electric shock at 1.8kV for 5ms, then immediately add 1mL liquid LB containing 0.5% (M / M) glucose for culture Incubate at 42°C for 5 minutes, spread on solid LB medium containing 100 μg / mL ampicillin and 35 μg / mL kanamycin, and culture at 30°C for 36 hours. After the total DNA was extracted from the tra...

Embodiment 3

[0041] The tertiary fermentation example of embodiment 3L-lysine

[0042] First-level preparation: the coryneform bacterium engineered bacterium that the transformation of embodiment 2 has pMS2-dap plasmid inserts 20 cubic meters of fermentors with 0.5% inoculum (the medium formula wherein is: 600 kilograms of glucose, 200 kilograms of sugarcane molasses , corn steep liquor 520 kg, KH 2 PO 4 45 kg, MgSO 4 ·7H 2 O 7 kg, FeSO 4 ·7H 2 O 0.5 kg, MnSO 4 ·7H 2 O 0.5 kg, 12 grams of biotin, and 5 grams of folic acid, the volume was adjusted to 18 cubic meters with water), and cultured at 31° C. with saturated aeration for 10 hours to increase the cell density. During the period, the amount of viable bacteria accumulated with glucose was significantly better than that with sucrose.

[0043] Second-level preparation: the nutrient solution of 20 cubic meters of fermentors is injected into 50 cubic meters of fermentors (wherein the formula of the medium is the same as above) wit...

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Abstract

The invention provides a fermentation method for preparing L-lysine products, which comprises the steps: an L-lysine culture solution is produced through three-level culture fermentation, and fine particles are coated by using fractional palm fat after spray drying. In addition, the invention also provides products prepared by the method, and the retention rate of L-lysine is larger than 15% within 24 hours.

Description

technical field [0001] The invention belongs to the field of amino acid fermentation. Specifically, the invention relates to a method for preparing L-lysine products by fermentation. Coated with palm fat. In addition, the present invention also provides products produced by the method and the like. Background technique [0002] L-lysine is an important amino acid raw material, which can be used as condiment, food, feed additive, and also as an effective or auxiliary ingredient in health care products and medicines. It is widely used in food industry, feed industry, pharmaceutical industry and other chemical industries. in industry. At present, the production of L-lysine is mainly produced by fermentation of microorganisms, such as coryneform bacteria. The microorganisms used for fermentation production can be wild-type microorganisms, but most of them are auxotrophs, drug-resistant mutants and metabolic mutants with higher yields obtained through mutagenesis or genetic en...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23K1/16C12P13/08C12R1/15A23K20/142
Inventor 马吉银陈崇安孟刚曹洪程耀东刘鑫
Owner NINGXIA EPPEN BIOTECH
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