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Embedding-crosslinking method for preparing nicotinic acid by Gibberella immobilization bioconversion

A technology for biotransformation, Gibberella, applied in biochemical equipment and methods, microorganism-based methods, immobilized on/in organic carriers, etc., can solve the problems of active damage, high glutaraldehyde toxicity, etc.

Inactive Publication Date: 2014-03-12
JIANGNAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Glutaraldehyde is highly toxic and will cause great damage to the activity of microorganisms during the immobilization process

Method used

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  • Embedding-crosslinking method for preparing nicotinic acid by Gibberella immobilization bioconversion
  • Embedding-crosslinking method for preparing nicotinic acid by Gibberella immobilization bioconversion
  • Embedding-crosslinking method for preparing nicotinic acid by Gibberella immobilization bioconversion

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0013] Glutaraldehyde cross-linked immobilized cells

[0014] The method of immobilizing cells with polyvinyl alcohol-chitosan composite materials: the prepared bacterial liquid is centrifuged to obtain wet bacterial cells, after centrifugal washing with normal saline, 0.05 g of bacterial cells are suspended in normal saline to prepare 10 g / L bacterial suspension, add polyvinyl alcohol solution with a mass fraction of 3% and chitosan solution with a mass fraction of 6%, mix thoroughly and uniformly according to a certain proportion, and then inject it with a syringe steadily into a solution containing 8% (w / v) polysaccharide Sodium phosphate saturated boric acid solution was solidified at 0°C for 6 hours to obtain immobilized pellets, which were filtered through gauze and then washed with PBS buffer to remove unfixed cells and mixed with the solidification solution. After curing, the pellets were stored in normal saline. The immobilized pellets are milky white and about 2mm i...

example 2

[0024] Genipin cross-linked immobilized cells

[0025] According to the method in Example 1, an equal amount of 7 parts of two kinds of immobilized cells were respectively prepared, and each part of the immobilized cells was treated with different concentrations of genipin for 12 hours, and the treated immobilized cells were washed with physiological saline after cross-linking, and then The immobilized cells were placed in saline and stored at 4°C.

[0026] The conditions for determining the viability were as follows: the immobilized cells cross-linked by genipin were placed in 7 mL of PBS buffer solution with a pH of 7.4, and 7 mL of 100 mM 3-cyanopyridine was added thereto to form a 14 mL reaction system for 30 min. After the reaction was completed, the liquid phase was used to detect the conversion result.

[0027] Table 3 Different concentrations of genipin treated polyvinyl alcohol-chitosan immobilized cells

[0028]

[0029] Table 4 Sodium alginate-gelatin immobiliz...

example 3

[0033] Batch conversion of niacin using two immobilized cells treated with two crosslinkers

[0034] Batch experiments were carried out with 6 mL of two kinds of immobilized cells treated with a certain amount of genipin and glutaraldehyde, and then 7 mL of PBS buffer was added to it, and 7 mL of substrate 3-cyanopyridine solution with a concentration of 100 mM was added, and the temperature was 40 °C. , react for 40min. At the end of each batch of transformation, the immobilized cells were washed with PBS buffer three times and then put into a new transformation system for the next batch of reactions.

[0035] Table 5 Batch transformation comparison of glutaraldehyde crosslinked immobilized cells and genipin immobilized cells

[0036]

[0037] It can be seen from Table 5 that the sodium alginate-gelatin immobilized cells without cross-linking treatment can only carry out 17 batches of reactions, and the cells immobilized by glutaraldehyde cross-linking can be reused for 2...

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Abstract

The invention belongs to the field of application of bioengineering technology, and particularly relates to a method for preparing nicotinic acid by immobilizing nitrilase-producing Gibberella CA3-1 (Gibberella intermedia, CGMCC No. 4903) with polyvinyl alcohol-chitosan and sodium alginate-gelatin composite materials and crosslinking respectively with glutaric dialdehyde and genipin. The genipin is used for preparing immobilized cells for the first time to enhance the mechanical properties of the immobilized cells. The method comprises the following steps: immobilizing Gibberella CA3-1, adding a proper amount of 3-cyanopyridine, and reacting under the optimal reaction conditions to obtain the product nicotinic acid. The research detects that the chitosan-polyvinyl alcohol immobilized cells crosslinked by the genipin have better activity retentivity and reutilization property than immobilized cells crosslinked by glutaric dialdehyde, and have higher activity than the sodium alginate-gelatin composite material; the chitosan-polyvinyl alcohol immobilized cells crosslinked by the genipin can be reutilized repeatedly, have higher substrate tolerance, and can enhance the production capacity of unit thalli. The method provides technical support for continuous production of nicotinic acid from 3-cyanopyridine by using the Gibberella CA3-1.

Description

technical field [0001] The invention relates to a genipin cross-linked polyvinyl alcohol-chitosan composite material immobilized nitrilase producing bacteria --- Gibberella CA3-1 ( Gibberella intermedia, CGMCC No.4903) A method for biologically preparing niacin, which belongs to the technical field of bioengineering applications. Background technique [0002] Nicotinic acid, chemical name pyridine-3-carboxylic acid, also known as nicotine acid, is an essential chemical substance for animals. It participates in the oxidation-reduction of organic matter and is widely used in daily life. Niacin is also an important pharmaceutical raw material and chemical intermediate. Niacin can be used to synthesize many drugs for the treatment of various skin diseases, hypertension symptoms and coronary heart disease. It can also be used as a drug intermediate in the production of isoniazid and other drugs, and is also widely used in luminescent materials, dyes, feed, etc. Niacin has the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/12C12N11/10C12N11/08C12N11/04C12R1/645
Inventor 许正宏杨涛李恒陆震鸣史劲松钱建瑛孙文敬周强
Owner JIANGNAN UNIV
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