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Bacterial strain capable of secreting cellulase as well as cellulase extraction method and application thereof

A cellulase and high cellulose technology, applied in the biological field, can solve the problems of low enzyme activity and yield, and achieve the effects of high cellulose degradation activity, wide industrial application prospects, and wide sources.

Inactive Publication Date: 2012-12-05
WUHAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the low yield of enzyme activity has become the biggest obstacle to the industrialization of cellulase

Method used

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  • Bacterial strain capable of secreting cellulase as well as cellulase extraction method and application thereof
  • Bacterial strain capable of secreting cellulase as well as cellulase extraction method and application thereof
  • Bacterial strain capable of secreting cellulase as well as cellulase extraction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment １

[0043] Example 1: Screening of bacterial strains

[0044]Trichoderma virens ZY-01 of the present invention (Trichoderma virens ZY-01, preserved in the China Center for Type Culture Collection, the preservation number is CCTCC No: M 2012205, and the preservation date is June 5, 2012) is a strain belonging to fungi, The strain was isolated from soil and obtained by enrichment and screening from soil samples with the substrate carboxymethylcellulose sodium (CMC?Na) as the only carbon source.

[0045] The method for screening high-efficiency cellulase strains is:

[0046] (1) Weigh 10 g of soil sample (in this example, the soil is taken from the corn stalk crop field), add 90 mL of sterile water, and oscillate to form a suspension. Stand still to clarify the supernatant and set aside;

[0047] (2) Take 1 mL of the supernatant after standing still, inoculate it in Congo red selective medium under sterile conditions, and culture it at 30-40 ℃ for 3-5 days;

[0048] (3) After 3-5 ...

Embodiment 2

[0049] Embodiment 2: bacterial strain identification

[0050] (1) The isolated bacterial strain in embodiment 1 adopts CTAB method to extract its genomic DNA:

[0051] Take 10 mL of bacterial liquid in a centrifuge tube at 4 °C and centrifuge at 10,000 r / min for 10 min to obtain mycelium; add 4 mL of CTAB and 0.8 mL of mercaptoethanol to the mycelia, mix evenly, and put in a 1.5 mL centrifuge tube for 30 min in a water bath ; Add an equal volume of phenol / chloroform isoamyl alcohol (the volume ratio of chloroform / isoamyl alcohol is 24:1), shake well, and centrifuge at 12000 r / min at 4 °C for 10 min; add 2 μL RNase to the supernatant, After mixing, keep in a 37°C water bath for 20-30 min; after the water bath, divide the mixture into 1.5 mL centrifuge tubes, about 400 μL per tube; add 2 times the volume of absolute ethanol and 1 / 10 volume of NaAC solution to precipitate; After the precipitation was complete, the supernatant was discarded by centrifugation, washed 2-3 times wit...

Embodiment 3-5

[0055] Embodiment 3-5: bacterial strain fermentation produces enzyme

[0056] The bacterial strain obtained in Example 1 was transferred into a fermentation medium for fermentation to produce enzymes to obtain crude enzyme liquid. The fermentation medium components of the strains described in the examples mainly involve carbon sources, nitrogen sources, and inorganic salts. The carbon source in the fermentation medium is sodium carboxymethylcellulose (CMC?Na), the nitrogen source is yeast extract (OXOID, UK), and the inorganic salts are NaCl and MgSO 4 ?7H 2 O and K 2 HPO 4 . The specific steps are shown in Table 1:

[0057]

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Abstract

The invention discloses a bacterial strain capable of secreting cellulase as well as the cellulase extraction method and application thereof, and solves the problems of low enzyme activity and yield in the conventional cellulase producing bacterial strain. The bacterial strain has a collection name of trichoderma virens ZY-01, is collected in China Center For Type Culture Collection (CCTCC), has a collection number of CCTCC No: M 2012205 and has an 18s rDNA sequence as shown in SEQ ID NO.1. A method for separating the bacterial strain, a method for producing, purifying and extracting the cellulase and an application method for efficient degradation of bagasses and straws are described. The bacterial strain has the advantages of high enzyme activity, wide range of adapted reaction condition and simplification of preparation; and the secreted cellulase is high in cellulose degradation activity.

Description

technical field [0001] The invention relates to the biological field, in particular to a method for isolating a bacterial strain capable of secreting cellulase, the cellulase produced by the bacterial strain and its extraction method and application. Background technique [0002] Cellulose is the most widely distributed polysaccharide in nature. If it can be rationally utilized and degraded into usable sugars, it can solve the current food and energy problems faced by human society. Cellulase is the key enzyme for hydrolyzing cellulose into fermentable sugars. After cellulose is hydrolyzed, reducing sugars can be obtained, which can be further converted into bioenergy and other substances through fermentation processes. Adding cellulase to forage can increase the utilization rate of feed and increase the meat production rate of animals. At present, cellulase is also widely used in industrial fields such as textile and papermaking. [0003] Most of the fungi used to produc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N1/02C12Q1/04C12N9/42C12P19/14C12R1/885
Inventor 杨忠华赵燕陈庚华周卫侯亚利
Owner WUHAN UNIV OF SCI & TECH
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