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Paenibacillus d7 and its application in degrading cellulose

A technology of bacillus and cellulose, which is applied to the new strains of cellulose degradation and the application field of cellulose degradation, which can solve the problems of less research and achieve the effects of easy cultivation, mild degradation conditions and strong reproductive ability

Active Publication Date: 2019-11-29
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The invention uses the water extract of waste inferior tobacco leaves as raw material, uses sodium carboxymethyl cellulose as the only carbon source, adopts Congo red cellulose identification medium for primary screening, and then uses the liquid fermentation culture added with sodium carboxymethyl cellulose Based on re-screening, a cellulose-degrading bacterium Paenibacillus sp.D7 was screened from it. After research, it has the ability to degrade cellulose. At present, there are few researches on cellulase produced by Paenibacillus at home and abroad.

Method used

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  • Paenibacillus d7 and its application in degrading cellulose
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  • Paenibacillus d7 and its application in degrading cellulose

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Embodiment 1: Screening of cellulose degrading bacteria

[0034] Tobacco waste extract (tobacco waste extract, TWE for short), domestic existing papermaking method tobacco sheet production process (see Figure 14 shown), the specific preparation method is: add water to tobacco chips, tobacco powder or tobacco stems, extract at 55°C-65°C for 20min-80min, and the volume of water used is about 8 times the mass of raw materials. After solid-liquid separation, the solid part (which can be directly used as cellulose for the degradation of Paenibacillus D7) is made into a paper base after pulping, and the liquid part is sprayed or soaked on the paper base after concentration, wherein the liquid part (i.e. (extract of waste tobacco leaves) is concentrated until no liquid flows out, washed with water, and dried to obtain a concentrate.

[0035] Take 1ml of the above waste tobacco leaf extract, add water to dilute the waste tobacco leaf extract to 10 -4 、10 -5 、10 -6, spread ...

Embodiment 2

[0044] Example 2: Identification of cellulose-degrading bacteria

[0045] On the primary screening medium, three strains with larger transparent circles were selected and inserted into the Congo red cellulose screening medium for re-screening, and the strain with the highest enzyme activity was named D7, so the follow-up cellulase research Using D7 as the research object, the size of the transparent circle produced by D7 (see figure 1 As shown) the colony shape is round, milky white, with a moist surface, and the ratio of colony diameter to transparent ring diameter is 2:7. TEM observation results (see figure 2 shown), strain D7 was rod-shaped with flagella. Gram-negative, spore-forming (see image 3 shown). The results of the physiological and biochemical characteristics of the strain were: starch hydrolysis, nitrate reduction, contact enzyme test, cellulose hydrolysis were positive, while indole test, urea test, citrate, H 2 S test was negative.

[0046] The bacterial...

Embodiment 3

[0047] Example 3: Growth characteristics and enzyme production of Paenibacillus sp.D7

[0048] (1) Slant culture: inoculate Paenibacillus D7 into the slant culture medium, cultivate at 37° C. for 24 hours to obtain slant bacteria; the final concentration of the slant culture medium consists of: sodium chloride 10 g / L, peptone 10 g / L, Yeast extract 5g / L, agar 18g / L, solvent is water, pH is 7.0.

[0049] (2) Seed culture: inoculate the slant bacteria in step (1) into the seed medium, cultivate at 37°C for 24 hours, and obtain the seed liquid; the final concentration of the seed medium consists of: sodium chloride 10g / L, peptone 10g / L L, yeast extract 5g / L, solvent is water, pH is 7.0.

[0050] (3) Fermentation culture: inoculate the seed liquid into the fermentation medium with an inoculum volume concentration of 1%, and cultivate it at 37° C. and 180 r / min for 24 hours to obtain a fermentation liquid. Sampling was taken to determine the dry weight of the bacteria, and the sta...

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Abstract

The invention discloses Bacillus genus D7 and its application in degrading cellulose. The strain of the invention can degrade sodium carboxymethyl cellulose (CMC-Na), cellulose of water extraction residue of waste inferior tobacco leaves and Cellulose materials such as tobacco leaves and stems, and the cellulase produced by the bacteria has the characteristics of high temperature resistance of 60°C and a wide range of catalytic substrate pH4-8. It has tolerance; compared with physicochemical and mechanical methods to degrade cellulose, the degradation conditions of the bacteria are milder; compared with fungi, bacteria have stronger reproductive ability, are easy to cultivate, and can be gradually enlarged, and industrial applications are expected to be realized in the later stage; The bacterial species of cellulase include Bacillus amyloliquefaciens, Clostridium thermobacterium, Paenibacillus and the like.

Description

[0001] (1) Technical field [0002] The invention relates to a kind of cellulose degradation, in particular to a new cellulose-degrading bacterial strain-Paenibacillus sp. D7 (Paenibacillus sp. D7) and its application in cellulose degradation. [0003] (2) Background technology [0004] Cellulose is the most extensive and abundant renewable carbon source on the earth. It can be degraded by cellulase into glucose, and then further converted into food, beverage or ethanol, etc., which can alleviate the food crisis or energy crisis. But cellulose is a macromolecular polysaccharide composed of glucose, usually combined with hemicellulose, pectin and lignin, insoluble in water and general organic solvents. At present, there are mainly the following methods for the degradation of cellulose: the first is acid-base degradation, using concentrated sulfuric acid or lye to degrade, but the recovery of acid-base has become a big problem, resulting in waste of acid-base; the second is mecha...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/42C12R1/01
CPCC12N9/2437C12Y302/01004C12N1/205C12R2001/01
Inventor 钟卫鸿郑艳红戴芸芸朱成云
Owner ZHEJIANG UNIV OF TECH
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