Biological preparation method of nicotinic acid, correlative strain and method for inducing nitrilase thereof

A nitrilase and hydrolase technology, applied in the field of microbiology, can solve the problems of many by-products, complicated processes, poor safety, etc., and achieve the effects of safe and simple production and no environmental pollution

Inactive Publication Date: 2009-11-11
EAST CHINA UNIV OF SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] At present, the industrial production methods of nicotinic acid include nitric acid oxidation method, potassium permanganate oxidation method, ammonia oxidation method, electrolytic oxidation method, quinoline oxidation method, etc. , but these traditional chemical indust

Method used

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  • Biological preparation method of nicotinic acid, correlative strain and method for inducing nitrilase thereof
  • Biological preparation method of nicotinic acid, correlative strain and method for inducing nitrilase thereof
  • Biological preparation method of nicotinic acid, correlative strain and method for inducing nitrilase thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] The cultivation of embodiment 1 rhododendron bacterium

[0050] Take the strain stored in a freeze-dried tube of Rhodobacter sphaericus, inoculate it on the slope of LB agar and cultivate it at 30°C for 24 hours.

[0051] Seed medium composition (pH is finally adjusted to 6-8):

[0052] Element

Embodiment 2

[0053] Induction of embodiment 2 nitrilase

[0054] Take a certain amount of the bacterial solution cultured in Example 1, inoculate it into the induction medium, add an appropriate amount of inducer at the same time, induce it at 30°C for 24 hours, collect the bacterial cells by centrifugation, and use NaH pH7.0 2 PO 4 / Na 2 HPO 4 The cells were washed twice with the buffer, and then suspended with an appropriate amount of buffer.

[0055] Induction medium (final pH adjusted to 6-8):

[0056] Element

Dose (g / L)

Glycerin

3.3

K H 2 PO 4

2

NaCl

1

MgSO 4

0.2

FeSO 4

0.03

[0057] The concentration of the inducer in the induction process is 10mM, and the condition of the induction process is the same as when cultivating the seeds, and the induction effects of different inducers are shown in the following table:

[0058] inducer

Embodiment 3

[0059] The biological preparation of embodiment 3 nicotinic acid

[0060] Add 1M 3-cyanopyridine to the resting cells suspended in 0.018g / ml (dry weight) induced by 10mM acetonitrile prepared in Example 2, so that the final concentration is 0.25M, transform in a shake flask at 30°C for 16h , using the HPLC method disclosed in Journal of Molecular Catalysis (B): Enzymatic 29 (2004) 227-232 to test the amount of nicotinic acid generated, the conversion rate of nicotinic acid from the substrate is 95%.

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Abstract

The invention provides a Rhodobacter sphaeroides (the preservation number is CCTCC M209065) for producing nitrilase, which can be cultured and generate the nitrilase in a general rich medium, wherein a nitrilase inducer is adopted to perform induction culture so that the nitrilase production level can be improved, and a nicotinic acid can be generated by biotransformation of 3-cyanopyridine by using a grown thallus. The nitrilase inducer is the 3-cyanopyridine, a 3-cyanopyridine structural analogue, a short-chain fatty nitrile compound, a short-chain fatty acid amide compound, an aromatic amide compound, an aromatic carboxylic acid compound, a short-chain fatty acid compound, urea, or no use of the inducer, and the concentration of the nitrilase inducer is between 1 and 200 mM. The method for preparing the nicotinic acid through the biotransformation has the advantages of safe and convenient production, environmental protection, fewer byproducts and high transformation rate.

Description

technical field [0001] The present invention relates to the technical field of microbiology, more specifically, to the technical field of using microorganisms to prepare nicotinic acid, in particular to a nicotinic acid biotransformation preparation method, related bacterial strains and a nitrilase induction method thereof. Background technique [0002] Nicotinic acid, the chemical name is pyridine 3-carboxylic acid, also known as nicotinic acid, the structural formula is The English name is 3-pyridine carboxylic acid or nicotinic acid, commonly known as vitamin B5, which belongs to the vitamin B family, and together with nicotinic acid, it is called vitamin PP. Niacin is mainly found in animal liver, kidney, muscle, yeast, wheat bran, rice bran, peanuts, fruits and vegetables, with a content ranging from 0.7 to 490 mg / kg. The appearance is white or light yellow crystal or crystalline powder, it is odorless, and the pH of 1% aqueous solution is 3.0-4.0. Its molecular weig...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/78C12P17/12C12R1/01
Inventor 王学东魏东芝张鲁嘉张建国杨春生王涛马成兵
Owner EAST CHINA UNIV OF SCI & TECH
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