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Fermentation medium and method for fermenting nicotinamide monoucleotide (NMN) transferase by same

A fermentation medium and fermentation method technology, applied in the directions of microorganism-based methods, transferases, biochemical equipment and methods, etc., can solve the problems of inability to meet the needs of industrial development, low enzyme production capacity and catalytic capacity, etc., and achieve cost Low, easy to operate, simple ingredients

Inactive Publication Date: 2016-06-22
UNIV OF SHANGHAI FOR SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the method of synthesizing NMN transferase is mainly biosynthesis, and traditional screening and mutagenesis techniques can be used to obtain some strains of NMN transferase with higher enzymatic activity, such as Staphylococcus aureus, Escherichia coli, extreme thermophilic Archaea, etc., however, these strains have low enzyme-producing and catalytic capabilities, which cannot meet the needs of industrial development

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] 1. Fermentation medium

[0018] The fermentation medium of this embodiment contains: yeast powder with a concentration of 20g / L, tryptone with a concentration of 8g / L, glycerol with a volume fraction of 4.5%, and MgSO with a concentration of 0.2g / L 4 , Yeast extract, NH concentration of 1.0g / L 4 Cl, NaCl with a concentration of 0.6g / L, KH with a concentration of 2g / L 2 PO 4 , K with a concentration of 9g / L 2 HPO 4 And double distilled water.

[0019] 2. Fermentation method

[0020] Using the fermentation medium with the component content of this embodiment to ferment nicotinamide mononucleotide adenosine (NMN) transferase, including the following steps:

[0021] Step 1: Streak E. coli into solid medium (preparation of solid medium: Dissolve LB medium with a concentration of 25g / L and agar with a concentration of 20g / L in a 500ml Erlenmeyer flask with double distilled water , Add NaOH to adjust the pH to 7.0, autoclave at 121°C for 20min), culture at 37°C for 12-18h to obtain a ...

Embodiment 2

[0033] 1. Fermentation medium

[0034] The fermentation medium of this embodiment contains: yeast powder with a concentration of 30g / L, tryptone with a concentration of 12g / L, glycerol with a volume fraction of 3%, and MgSO with a concentration of 0.3g / L 4 , Yeast extract, NH concentration of 1.5g / L 4 Cl, NaCl with a concentration of 0.4g / L, KH with a concentration of 2g / L 2 PO 4 , K with a concentration of 9g / L 2 HPO 4 And double distilled water.

[0035] 2. Fermentation method

[0036] Using the fermentation medium with the component content of this embodiment to ferment nicotinamide mononucleotide adenosine (NMN) transferase, including the following steps:

[0037] Step 1: Streak E. coli into solid medium (preparation of solid medium: Dissolve LB medium with a concentration of 25g / L and agar with a concentration of 20g / L in a 500ml Erlenmeyer flask with double distilled water , Add NaOH to adjust the pH to 7.0, autoclave at 121°C for 20min), culture at 37°C for 12-18h to obtain a s...

Embodiment 3

[0049] 1. Fermentation medium

[0050] The fermentation medium of this embodiment contains: yeast powder with a concentration of 20g / L, tryptone with a concentration of 8g / L, glycerol with a volume fraction of 3%, and MgSO with a concentration of 0.2g / L 4 , Yeast extract, NH concentration of 1.5g / L 4 Cl, NaCl with a concentration of 0.6g / L, KH with a concentration of 3.0g / L 2 PO 4 , K with a concentration of 9.0g / L 2 HPO 4 And double distilled water.

[0051] 2. Fermentation method

[0052] Using the fermentation medium with the component content of this embodiment to ferment nicotinamide mononucleotide adenosine (NMN) transferase, including the following steps:

[0053] Step 1: Streak E. coli into solid medium (preparation of solid medium: Dissolve LB medium with a concentration of 25g / L and agar with a concentration of 20g / L in a 500ml Erlenmeyer flask with double distilled water , Add NaOH to adjust the pH to 7.0, autoclave at 121°C for 20min), culture at 37°C for 12-18h to obtain ...

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Abstract

The invention provides a fermentation medium and a method for fermenting nicotinamide monoucleotide (NMN) transferase by the same.The method includes the following steps that 1, a solid medium containing Escherichia coli is prepared; 2, activation liquor is prepared; 3, Escherichia coli is inoculated into the activation liquor, and kanamycin sulfate is added to obtain liquor containing activated Escherichia coli thalli; 4, the liquor containing thalli is inoculated into a sterilized fermentation medium for fermentation cultivation, an inductive agent IPTG is added for inductive cultivation, and fermentation liquor containing NMN transferase is collected; 5, the fermentation liquor is centrifuged, sediment is taken, and then Escherichia coli wet thalli containing NMN transferase are obtained.The fermentation medium has simple components, is low in cost and has broad application prospects, and the fermentation method is simple, practical and easy to implement.

Description

Technical field [0001] The invention relates to the field of microbial fermentation engineering, in particular to a fermentation medium and a method for fermenting NMN transferase with the fermentation medium. Background technique [0002] Studies have found that NMN transferase is widely present in cells of animals, plants, prokaryotes and eukaryotes. At present, there are three main subtypes of NMN transferase in cells, which are Nmnat1, Nmnat2 and Nmnat3, and its molecular weight, properties and characteristics vary with its distribution in tissue cells. Nicotinamide mononucleotide (NMN) plays an important role in human cell energy production. It is involved in the synthesis of intracellular nicotinamide adenine dinucleotide (NAD, an important coenzyme for cell energy conversion). In all living biological tissues, NMN transferase is an indispensable biological enzyme for the synthesis of NAD. Studies have shown that it is very important for the survival of prokaryotic cells a...

Claims

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Application Information

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IPC IPC(8): C12N9/12C12R1/19
CPCC12N9/1241C12Y207/07001
Inventor 李红梅段琳琳袁飞飞亢涵
Owner UNIV OF SHANGHAI FOR SCI & TECH
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