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Cellulose degrading bacterium and application thereof

A technology of cellulose degradation and cellulose, applied in the direction of bacteria, microorganisms, biochemical equipment and methods, etc., can solve the problem of low efficiency of straw degradation

Active Publication Date: 2022-04-08
NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most commercial cellulases on the market are secreted by Trichoderma and Aspergillus, such as Trichoderma reesei, Trichoderma konshii and Aspergillus niger; The efficiency of degrading straw is low, so finding a highly efficient cellulose-degrading bacteria is of great significance for improving the efficiency of straw degradation

Method used

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  • Cellulose degrading bacterium and application thereof
  • Cellulose degrading bacterium and application thereof
  • Cellulose degrading bacterium and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0020] Weigh 10g of straw compost samples (composted from the Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences) in different sampling periods, add them into 90ml of sterilized ultrapure water, shake at 30°C and 180rpm for 30min, mix well and then let stand, take 1ml of suspension The solution was serially diluted, and 70 μl of the diluted suspension was spread on the CMC solid medium, cultured in a 30°C incubator for 5 days, and the colonies on the CMC solid medium were streaked and purified repeatedly for 5 to 8 times, and the purified single The strains were cultured on CMC solid medium for 3 days and then stained with Congo red. After washing with 1mol / L NaCl, those with transparent circles on the plate were possible cellulose-degrading bacteria, and one strain with obvious cellulose-degrading transparent circles was screened. strain, numbered as strain WF-10.

[0021] Molecular biological identification of strain WF-10:

[0022] The culture of...

Embodiment 2

[0025] Determination of Cellulase Activity of Streptomyces ansochromogenes WF-10

[0026] Inoculate the Streptomyces ansochromogenes WF-10 into the fermentation enzyme production medium (fermentation enzyme production medium: CMC-Na 10g, NaCl 0.1g, KH 2 PO 4 1.0g, NaNO 3 2.5g, MgSO 4 ·7H 2 O 0.3g, CaCl 2 0.1g, FeCl 3 0.01g, distilled water 1000mL), shake culture at 30°C and 200rpm, take a sample every 24h, centrifuge at 8000rpm for 5min, and the supernatant is the crude enzyme solution.

[0027] A. Determination of endoglucanase activity: Take 4 clean test tubes of the same specification, 1 test tube as blank, and the remaining 3 as test tubes. Accurately weigh 1.5mL of 1% CMC-Na standard solution, add to 4 test tubes respectively, add 0.5mL diluted crude enzyme solution to 3 test tubes respectively, shake well and place 4 test tubes at 50℃ React in a water bath for 30 minutes. After taking it out, add 0.5mL of boiled inactivated diluted crude enzyme solution to th...

Embodiment 3

[0033] To verify the high efficiency and practical applicability of Streptomyces ansochromogenes WF-10, liquid fermentation and solid fermentation experiments were carried out.

[0034] 1. Degradation of straw by liquid fermentation: Inoculate 5 mL of Streptomyces ansochromogenes WF-10 bacterial liquid into the optimized liquid fermentation medium (optimized liquid fermentation medium formula: 15 g of corn straw powder, NaCl 0.1 g.KH 2 PO 4 1.0g, (NH 4 ) 2 SO 4 4g, MgSO 4 ·7H 2 O 0.3g, CaCl 2 0.1g, FeCl 3 0.01g, distilled water 1000mL), with the liquid fermentation medium not inoculated with bacteria liquid as a control, after culturing for 7 days at 30°C and 180rpm, the supernatant was discarded by centrifugation, and the straw was washed repeatedly with dilute acid solution, and then washed with distilled water After centrifugation, the relative degradation rate of straw was determined by weight loss method.

[0035] 2. Degradation of straw by solid-state fermen...

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Abstract

The invention discloses a cellulose degrading bacterium and application thereof, and relates to a cellulose degrading microorganism and application thereof. According to the present invention, the cellulose degrading bacterium is Streptomyces ansochromogenes WF-10, and is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.23720; the invention also discloses an application of the cellulose degrading bacterium in cellulose biodegradation. The strain disclosed by the invention can grow on a CMC (Carboxymethyl Cellulose) plate taking sodium carboxymethyl cellulose as a unique carbon source, and an obvious cellulose decomposition transparent ring appears around a bacterial colony after the bacterial colony is dyed by Congo red; according to the invention, the cellulase can secrete four types of cellulase, including filter paper enzyme, endo-beta-1, 4-glucanase, exo-beta-1, 4-glucanase and beta-glucosidase; the capability of degrading natural cellulose is realized.

Description

technical field [0001] The invention relates to a cellulose degrading microorganism and its application. Background technique [0002] my country is a large agricultural country with abundant agricultural wastes, especially corn stalks. According to statistics, from 2014 to 2018, the average annual output of straw in my country was about 653.866 million tons, of which corn straw accounted for about 45% of the total straw. Corn stalks contain a variety of nutrients, including C, N, P, K and trace elements, and are important biomass resources. [0003] The main component of corn stover is lignocellulose, which is a rich renewable resource that can be converted into a variety of bioenergy and chemical products. Straw returning to the field is the main measure for the rational use of black soil protection. The key technology requires that the straw can be effectively degraded in the soil and transformed into soil organic matter. How to realize the effective and rapid degradat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10C12R1/465
Inventor 田春杰王新光王恩泽
Owner NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
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