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Bacteriostatic chitin enzymolysis method

A technology of chitinase and chitin, applied in the biological field, can solve the problems of dangerous operation, expensive equipment, and restrictions on the development of the biological sugar industry, and achieve the effects of avoiding pollution, improving enzymatic hydrolysis efficiency, and shortening the reaction cycle

Inactive Publication Date: 2015-04-22
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the above-mentioned technologies are relatively mature, there are disadvantages of expensive equipment and dangerous operation, which limit the development of biological sugar industry

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Step 1, microbial fermentation to produce chitinase

[0029] by Chitinolyticbacter meiyuanensis The SYBC-H1 strain is a chitinase-producing strain, and the seed medium is (2 g / L glucose, 2 g / L peptone, 0.7 g / L KH 2 PO 4 , 0.3 g / L K 2 HPO 4 ·3H 2 O, 0.4 g / L MgSO 4 ·7H 2 O, pH 7.0), incubated at 37°C, 200 rpm for 12 h. The seed liquid was inserted into a 7.5 L fermenter equipped with an enzyme-producing fermentation medium at an inoculum volume of 8%, with a liquid volume of 4 L, and the enzyme-producing medium was (3 g / L powder chitin, 3 g / L glucose, 3 g / L beef extract, 0.7 g / L KH 2 PO 4 , 0.3 g / L K 2 HPO 4 ·3H 2 O, 0.5 g / L MgSO 4 ·7H 2 O) The culture temperature was 30 °C, the rotation speed was 200 rpm, the aeration rate was 1 vvm, the initial pH was 7.5, and the fermentation was terminated after 72 h.

[0030] The fermentation broth was centrifuged at 8000 g, the cells were discarded, and the supernatant was collected and then precipitated and reconstit...

Embodiment 2

[0036] Step 1, microbial fermentation to produce chitinase

[0037] by Chitinibacter The sp.GC72 strain is a chitinase-producing strain, and the seed medium is (4 g / L glucose, 4 g / L peptone, 0.7 g / L KH 2 PO 4 , 0.3 g / L K 2 HPO 4 ·3H 2 O, 0.4 g / L MgSO 4 ·7H 2 O, pH 7.0), cultured at 37 °C and 200 rpm for 12 h, the seed liquid was placed in a 7.5 L fermenter with an enzyme-producing fermentation medium at a volume fraction of 6% inoculum, with a liquid volume of 4 L, and the enzyme-producing medium is (4 g / L powdered chitin, 2 g / L glucose, 4 g / L beef extract, 0.7 g / L KH 2 PO 4 , 0.3 g / L K 2 HPO 4 ·3H 2 O, 0.5 g / L MgSO 4 ·7H 2 O) The culture temperature was 30 °C, the rotation speed was 200 rpm, the ventilation rate was 1 vvm, the initial pH was 7.5, and the fermentation broth was collected after 72 h.

[0038]The fermentation broth was centrifuged at 8000 g, the cells were discarded, and the supernatant was collected and then precipitated and reconstituted with am...

Embodiment 3

[0044] Step 1, microbial fermentation to produce chitinase

[0045] by Chitinibacter The sp.GC72 strain is a chitinase-producing strain, and the seed medium is (4 g / L glucose, 4 g / L peptone, 0.7 g / L KH 2 PO 4 , 0.3 g / L K 2 HPO 4 ·3H 2 O, 0.4 g / L MgSO 4 ·7H 2 O, pH 7.0), cultured at 37 °C and 200 rpm for 12 h, the seed solution was placed in a 7.5 L fermenter with an enzyme-producing fermentation medium at a volume fraction of 6% inoculum, with a liquid volume of 4 L, and the enzyme-producing medium is (4 g / L powdered chitin, 2 g / L glucose, 4 g / L beef extract, 0.7 g / L KH 2 PO 4 , 0.3 g / L K 2 HPO 4 ·3H 2 O, 0.5 g / L MgSO 4 ·7H 2 O) The culture temperature was 30 °C, the rotation speed was 200 rpm, the ventilation rate was 1 vvm, the initial pH was 7.5, and the fermentation broth was collected after 72 h.

[0046] The fermentation broth was centrifuged at 8000 g, the cells were discarded, and the supernatant was collected and then precipitated and reconstituted with...

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Abstract

The invention discloses a bacteriostatic chitin enzymolysis method which comprises the following steps: 1, preparation of chitinase through microbial fermentation: by taking a chitinase preparation microorganism as a test bacterium, fermenting to prepare chitinase, obtaining chitinase crude enzyme after the fermentation is finished, and purifying, dialyzing and concentrating to obtain chitin degrading enzyme; 2, enzymolysis of chitin: putting the chitin degrading enzyme in a reaction kettle, adding chitin powder, a metal ion activating agent, an auxiliary enzyme preparation and an organic reagent, and heating at constant temperature and stirring to obtain enzymatic hydrolysate when the chitin powder is completely degraded; and 3, recovery of product: centrifuging the enzymatic hydrolysate to obtain a biphasic system, wherein the upper system is an organic system and the lower system is a water system, adsorbing by adsorption resins, eluting, collecting the eluent and treating the eluent to obtain the product. According to the bacteriostatic chitin enzymolysis method, the raw materials can be repeatedly utilized and the conversion process is sterile so that high yield can be obtained when the energy consumption is reduced; and the degraded product is favorable in biological activity so that remarkable economic benefit is provided.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a bacteriostatic chitin enzymatic hydrolysis method. Background technique [0002] The sugar industry is the basic industry of the food industry, as well as the raw material industry for chemical, fermentation, pharmaceutical and other products. Statistics show that from 2012 to 2013, my country's sugar production reached 13.068 million tons. Among them, as a sunrise industry in the field of food biotechnology, the functional sugar and its derivatives industry has developed rapidly in recent years. As a polysaccharide with natural content second only to cellulose, chitin, its degradation products and its derivatives are widely used in many fields such as food, medicine, environmental protection, agriculture, and daily chemicals, and have formed a huge market demand. However, the production of chitin degradation products and their derivatives is mostly chemical methods. The reaction...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12P19/26C12R1/01
CPCC12N9/2442C12P19/26C12Y302/01014
Inventor 陈可泉高聪张阿磊何珣欧阳平凯
Owner NANJING UNIV OF TECH
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