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Method of immobilizing gibberella and using gibberella for biological transformation to prepare nicotinic acid

A technology for Gibberella, immobilized pellets for use in microorganism-based methods, biochemical equipment and methods, immobilization on/in organic carriers, etc.

Active Publication Date: 2013-04-24
JIANGNAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, there have never been any patents and literature reports on the use of nitrilase-producing filamentous fungi to prepare immobilized cells for the biotransformation of nitriles

Method used

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  • Method of immobilizing gibberella and using gibberella for biological transformation to prepare nicotinic acid
  • Method of immobilizing gibberella and using gibberella for biological transformation to prepare nicotinic acid
  • Method of immobilizing gibberella and using gibberella for biological transformation to prepare nicotinic acid

Examples

Experimental program
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Effect test

example 1

[0016] 1. Preparation of hydrolase-producing bacteria liquid

[0017] Liquid medium: Potassium dihydrogen phosphate 2.72g, ferrous sulfate 0.0139g, glucose 10g, yeast powder 7.5g, sodium chloride 1.16g, caprolactam 3.39g, add water to 1L, fully dissolve and adjust the pH value to 7.0.

[0018] Cultivation method: pick fungi from the slant strains with an inoculation loop, inoculate in the seed medium of the shake flask, fill 50mL liquid medium in a 500mL Erlenmeyer flask, and cultivate for 60 hours at 30°C and 220rpm.

[0019] 2. Bacteria immobilization

[0020] The prepared bacterial liquid was centrifuged to obtain the bacterial cells, and after centrifugal washing with normal saline, 0.03 bacteria were resuspended in normal saline to prepare a bacterial suspension of 9 g / L, to which was added alginic acid with a mass fraction of 5.5% Sodium solution, mix well and inject it steadily with a syringe until the mass fraction is 1% (w / v) CaCl 2 In the solut...

example 2

[0028] Add 7 mL of PBS buffer (containing 30 mM CaCl 2 ) by adding 7 mL of substrate 3-cyanopyridine with a concentration of 100 mM, pH 9.0, reacting for 35 min, and measuring the specific enzyme activity of the immobilized bacteria at different temperatures (see Table 2), the results show that when the reaction temperature is 40 °C The specific enzyme activity of the immobilized bacteria was the highest, so the optimum reaction temperature of the reaction system was determined to be 40°C.

[0029] temperature (°C) 25 30 35 40 45 50 55 specific enzyme activity 402.6 512.7 587.1 674.2 597.3 436.9 330.2

example 3

[0031] Add 7 mL of PBS buffer (containing 30 mM CaCl 2 ) by adding 7 mL of substrate 3-cyanopyridine with a concentration of 100 mM, reacting for 35 min at 40 ° C, and measuring the specific enzyme activity of the immobilized bacteria at different pH (see Table 3), the results show that the immobilized bacteria at pH 9.0 The specific enzyme activity of the body is the highest, so the optimum pH of the reaction system is 9.0.

[0032] pH value 6.0 7.0 8.0 9.0 10.0 11.0 specific enzyme activity 366.8 493.7 568.1 642.4 532．6 452.7

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Abstract

The invention belongs to the application field of biology engineering technology, and particularly relates to a method of immobilizating gibberella CA3-1(Gibberella intermedia) with a preserved number of CGMCC No.4903 and using the gibberella for biological transformation to prepare nicotinic acid. The method of immobilizing the gibberella and using the gibberella for the biological transformation to prepare the nicotinic acid includes: immobilizing the gibberella CA3-1, adding a proper amount of 3-cyanopyridine, and choosing an optimum reaction condition for immobilizing thalluses to obtain a product which is the nicotinic acid. The method of immobilizing the gibberella and using the gibberella for the biological transformation to prepare the nicotinic acid strengthens tolerance of the thalluses to substrates after the thalluses are immobilized, improves production capacity of unit thalluses, simplifies separation processes of products greatly, increases reutilization times obviously, and provides a foundation for achieving technology of performing a biological transformation method by using the gibberella to prepare the nicotinic acid.

Description

technical field [0001] The present invention relates to a kind of Gibberella CA3-1 ( Gibberella intermedia , CGMCC No. 4903) immobilized and used for the preparation of nicotinic acid, which belongs to the technical field of bioengineering applications. Background technique [0002] Niacin belongs to vitamin B series compounds and is an indispensable nutrient for the human body. It has a wide range of effects in promoting normal human development, promoting metabolism, dilating blood vessels, and relieving spasm of peripheral blood vessels. At the same time, niacin is also an important pharmaceutical intermediate and chemical intermediate, which can be used as a pharmaceutical intermediate to produce isoniazid, nicothamide, and nicotinic acid inositol ester to treat skin diseases, hypertension, arteriosclerosis and coronary artery disease. Drugs for diseases such as heart disease. [0003] Traditional nicotinic acid production mainly adopts chemical synthesis methods, such...

Claims

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Application Information

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IPC IPC(8): C12P17/12C12N11/10C12N11/04C12R1/645
Inventor 史劲松李恒杨涛龚劲松许正宏孙文敬周强
Owner JIANGNAN UNIV
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