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A cellulase-producing Aspergillus niger and its application

A technology of cellulase and Aspergillus niger, applied in the field of halophilic Aspergillus niger and its cultivation

Active Publication Date: 2018-02-02
深圳市中荃科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, most of the studies on cellulase-producing strains focus on conventional low-salt microorganisms, and few studies on cellulase-producing microorganisms in salt lakes have been reported so far. Only Vibrio sp.G21 has been reported (Gao Z et al.2010) , Chromohalobacter sp.TPSV 101(Prakasha et al.2012), Haloarcula sp.G10, Haloarcula sp.LLSG7(Li X et al.2013), Pestalotiopsis sp.NCi6(Arfi Y et al.2013), etc.

Method used

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  • A cellulase-producing Aspergillus niger and its application
  • A cellulase-producing Aspergillus niger and its application
  • A cellulase-producing Aspergillus niger and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] The screening of the optimal carbon source of embodiment 1 Aspergillus niger producing cellulase fermentation medium

[0055] ① Inoculate the seed solution at 10% inoculum amount in the fermentation medium whose carbon source is corncob powder, bran, cellulose, sawdust, and rice bran, at 28°C, 160r min -1 Conditioned for 72h.

[0056] ② Make the fermentation broth 4000r·min -1 Centrifuge for 10 min to obtain the supernatant crude enzyme solution. Take 1 mL of the crude enzyme solution and add 9 mL of pH 4.8, 0.05 mol / L citric acid buffer to prepare a diluted enzyme solution. Take two 20mL test tubes, add 0.5mL diluted enzyme solution and 1.5mL 0.5% CMC solution to one test tube, add 2.0mL 0.5% CMC solution to the other control tube, and do 3 replicates for each tube of the diluted enzyme solution test tube.

[0057] ③ 50 ℃ water bath for 30 minutes, add 3mL DNS reagent.

[0058] ④Bath in boiling water for 5 minutes, take it out and immediately put it in ice water to ...

Embodiment 2

[0060] The optimal nitrogen source screening of embodiment 2 Aspergillus niger producing cellulase fermentation medium

[0061] ①Determine the optimal carbon source, use (NH4)2SO4, soybean powder, beef extract, yeast extract, and peptone as nitrogen sources, inoculate the seed solution at 10% inoculum in the fermentation medium, 28°C, 160r min -1 Conditioned for 72h.

[0062] 2.-5. steps are the same as in Example 1. To draw a histogram of enzyme activity values ​​against different nitrogen sources, see Figure 4 , the study of enzymatic properties showed that the best nitrogen source for the strain to produce cellulase fermentation medium was peptone with a mass fraction of 0.5%.

Embodiment 3

[0063] The optimal temperature screening of embodiment 3 Aspergillus niger producing cellulase fermentation culture

[0064] ①Determine the optimal carbon source and nitrogen source medium, inoculate the seed solution in the fermentation medium at 10% inoculation amount, and inoculate them at 24°C, 26°C, 28°C, 30°C, 32°C, 160r·min -1 Conditioned for 72h.

[0065] 2.-5. steps are the same as in Example 1. To draw a line graph of enzyme activity values ​​against different temperatures, see Figure 5 , the study of enzymatic properties showed that the optimum temperature for the strain to produce cellulase fermentation medium was 28°C.

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Abstract

The invention discloses a cellulase-producing halophilic Aspergillus niger and application thereof, belonging to the field of microorganisms. The preservation number of the disclosed Aspergillus niger strain is CGMCC9404. The Aspergillus niger strain was isolated from the water deposits of Huama Salt Lake Lake, Dingbian County, Yulin City, Shaanxi Province, Shaanxi Province, China. The strain was identified as Aspergillus niger by morphology and molecular biology. It can be cultured at 28°C for 5 days in a medium with a carbon source of 2% corncob flour, a nitrogen source of 0.5% peptone, an initial pH of 5.5, a spore suspension inoculation volume fraction of 20%, and a salt concentration of 2.5%. , reaching the highest enzyme activity. And the strain can produce cellulase under the condition that the salt concentration is 0%-15%.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a cellulase-producing halophilic Aspergillus niger and its cultivation method and application. Background technique [0002] Cellulose is one of the most abundant renewable resources on earth. It is a polysaccharide composed of β-D-1,4 glucosidic bonds. Its molecular weight can reach hundreds of thousands or even millions, and it cannot be directly used by microbial cells. . Cellulase is a general term for a class of complex enzymes that biodegrade cellulose containing β-1,4 glycosidic bonds to generate glucose. It is mainly composed of three hydrolytic enzymes, namely β-glucansidase and cellobiohydrolase and β-glucosidase. First, β-glucosidase acts on the non-crystalline region of microfibrils, exposing many ends for the action of exo-type enzymes. Cellobiohydrolase decomposes from the non-reducing ends in turn to produce cellobiose, and then partially degrades the Ce...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C02F3/34C09K17/00C12R1/685
CPCC02F3/347C02F2103/28C02F2103/30C09K17/00C12N1/14C12N1/145C12R2001/685
Inventor 刘开辉张波丁小维陈文强邓百万杨方玉
Owner 深圳市中荃科技有限公司
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