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Liquid fermentation method for producing salt-tolerant cellulase from sea aspergillus niger

A marine Aspergillus niger, liquid fermentation technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, enzymes, etc., to achieve the effects of stable enzyme activity, easy control and amplification, and simple medium

Inactive Publication Date: 2013-06-05
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cellulase-producing microorganisms have been screened from the ocean, mainly bacteria and yeasts, but molds with rich enzymes, high expression levels, and strong secretion capabilities have rarely been reported

Method used

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  • Liquid fermentation method for producing salt-tolerant cellulase from sea aspergillus niger
  • Liquid fermentation method for producing salt-tolerant cellulase from sea aspergillus niger
  • Liquid fermentation method for producing salt-tolerant cellulase from sea aspergillus niger

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1) A marine Aspergillus niger was screened from the soil at a depth of 10m off the coast of the East China Sea;

[0035] 2) Aspergillus niger was inoculated on PDA medium, cultured at 40°C for 60 hours to obtain slant spores, and the slant spores were stored at 4°C;

[0036] 3) The slant spores were inoculated in PDA-1 medium, and cultured at 40° C. for 60 hours to obtain activated spores;

[0037] 4) Put 60g of eggplant bottle slant culture medium into a 250ml eggplant bottle and compact it into a slant, seal the mouth of the bottle with 8 layers of gauze to obtain the slant of the eggplant bottle, inoculate the activated spores on the slant of the eggplant bottle, and culture at 42°C for 74 hours to obtain Eggplant bottle slant spores for expansion; the composition of eggplant bottle slant expansion culture medium is: three-layer gauze filtrate 80mL or 100mL or 120mL of potato artificial seawater boiled for 30min, glucose 2g, straw powder 50g; the composition of artif...

Embodiment 2

[0041] 1) A marine Aspergillus niger was screened from the soil at a depth of 10m off the coast of the East China Sea;

[0042] 2) Aspergillus niger was inoculated on PDA medium, cultured at 40°C for 60 hours to obtain slant spores, and the slant spores were stored at 4°C;

[0043] 3) The slant spores were inoculated in PDA-1 medium, and cultured at 40° C. for 60 hours to obtain activated spores;

[0044] 4) Put 60g of eggplant bottle slant culture medium into a 250ml eggplant bottle and compact it into a slant, seal the mouth of the bottle with 8 layers of gauze to obtain the slant of the eggplant bottle, inoculate the activated spores on the slant of the eggplant bottle, and culture at 42°C for 74 hours to obtain Eggplant bottle slant spores for expansion; the composition of eggplant bottle slant expansion culture medium is: 100mL of three-layer gauze filtrate boiled in artificial seawater for 30min, 1g or 2g or 6g of glucose, 50g of straw powder; the composition of artifici...

Embodiment 3

[0047] 1) A marine Aspergillus niger was screened from the soil at a depth of 10m off the coast of the East China Sea;

[0048] 2) Aspergillus niger was inoculated on PDA medium, cultured at 40°C for 60 hours to obtain slant spores, and the slant spores were stored at 4°C;

[0049] 3) The slant spores were inoculated in PDA-1 medium, and cultured at 40° C. for 60 hours to obtain activated spores;

[0050] 4) Put 60g of eggplant bottle slant culture medium into a 250ml eggplant bottle and compact it into a slant, seal the mouth of the bottle with 8 layers of gauze to obtain the slant of the eggplant bottle, inoculate the activated spores on the slant of the eggplant bottle, and culture at 42°C for 74 hours to obtain Eggplant bottle slant spores for expansion; the composition of the eggplant bottle slant expansion culture medium is: three layers of gauze filtrate 100mL, glucose 2g, rice straw powder 10g or 50g; artificial seawater is composed of: NaCl 29.8g, MgSO 4 5.4g, KCl ...

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Abstract

The invention discloses a liquid fermentation method for producing salt-tolerant cellulase from sea aspergillus niger. The liquid fermentation method comprises the following steps: 1) screening soil in 10-20m depth of coastal area of the East China Sea to obtain a strain of sea aspergillus niger; 2) inoculating the sea aspergillus niger into a PDA (potato dextrose agar) culture medium, culturing at 32-40 DEG C for 60-90 hours to obtain slant spores, and preserving the slant spores at 4 DEG C; 3) inoculating the slant spores on a PDA-1 culture medium, and culturing at 32-40 DEG C for 60-90 hours to obtain activated spores; 4) inoculating the activated spores on a slant of an eggplant bottle, and culturing at 42 DEG C for 74 hours to obtain eggplant bottle slant spores subject to expanding culture; and 5) utilizing sterile artificial sea water to dilute the eggplant bottle slant spores obtained by expanding culture to 108 spores / ml, inoculating on a liquid fermentation enzyme productionculture medium, and fermenting for 5-10 days to obtain the cellulase. The liquid fermentation method has the advantages that the produced enzyme has high activity, stability, good salt tolerance and the like, and the produced cellulase can degrade cellulose under the condition of higher salt content, thereby laying a foundation for follow-up application of hydrolysate.

Description

technical field [0001] The invention relates to a liquid fermentation method for producing salt-tolerant cellulase from marine Aspergillus niger. Background technique [0002] Cellulosic materials hold great promise for the production of biobased products. There are a large number of cellulose material resources in nature, such as corn straw, rice straw, wheat straw, rape straw and so on. The rich cellulose contained in these substances can be degraded to obtain fermentable sugar-glucose, which can be used to produce a large number of bio-based products, such as bioethanol, butanol, citric acid, lactic acid, single-cell protein, etc. In particular, the use of cellulosic materials to produce bioenergy such as hydrogen and ethanol can alleviate the increasingly tense energy pressure, and has very broad prospects. [0003] A key step in the utilization of cellulosic material is its conversion to fermentable sugars. There are usually two types of methods to degrade cellulose:...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/42C12N1/14C12R1/685
Inventor 姚善泾薛栋升林东强刘杰凤
Owner ZHEJIANG UNIV
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