Bacillus subtilis and application thereof in sisal hemp degumming

A Bacillus subtilis, degumming technology, applied to the application of sisal degumming, in the field of Bacillus subtilis, to achieve the effects of low production cost, shortened degumming time and high yield

Inactive Publication Date: 2010-11-10
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The inventor of the research on sisal degumming has made a little progress at home and abroad. He proposed that B2 bacteria have certain sisal degumming ability (Chen Qing, Han Shuangyan, etc., Screening and identification of si

Method used

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  • Bacillus subtilis and application thereof in sisal hemp degumming
  • Bacillus subtilis and application thereof in sisal hemp degumming
  • Bacillus subtilis and application thereof in sisal hemp degumming

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: Enrichment and screening of sisal degumming bacteria

[0038]Red Star Farm in Zhanjiang City, Guangdong Province collected 9 kinds of soil samples from sisal production sites, weighed 1.00g each, added them to a triangular flask filled with 99mL sterile water and an appropriate amount of sterilized glass beads, oscillated evenly to obtain a bacterial suspension, and took 0.5mL Put the bacterial suspension into the beef peptone enrichment medium, cultivate it for 1 day, then inoculate the bacterial solution from the enrichment medium into the degumming medium, cultivate it at 30°C and 150r / min for 3 days, and observe whether there is degumming phenomenon .

[0039] In order to stabilize the degumming performance, domesticate the mixed flora with degumming effect, that is, cycle degumming, the specific method is as follows: the degumming liquid with degumming effect after 3 days of degumming is transferred to the enrichment medium for 1 day, and then transfe...

Embodiment 2

[0040] Embodiment 2: the taxonomic identification of bacterial strain

[0041] According to "Bergey's Bacteria Identification Manual" (Ninth Edition) and "Common Bacteria System Identification Manual", B2 bacteria grow in flocculent form in liquid medium, the cells are rod-shaped, mobile, and Gram staining is positive. There are spores, the spores are cylindrical or oval, and their physiological and biochemical characteristics are most consistent with Bacillus subtilis (Bacillus subtilis), and they do not grow at 50°C, but their colony morphology is different from the common Bacillus subtilis colony morphology. At the same time, its 16S rDNA sequence (submitted to GeneBank, No. EF639849) was homologously compared in GenBank, and the results showed that the strain B2 had a high homology with Bacillus subtilis, with a homology of 99%. Biological characteristics are: rod-shaped bacteria, positive Gram staining, flocculent growth in liquid medium, spores, columnar or oval, aerobic...

Embodiment 3

[0042] Embodiment 3: xylanase enzyme, pectinase activity assay in degumming

[0043] Since pectinase, hemicellulase (mainly xylanase) and cellulase degrade the glycosidic bonds of the substrate to generate products containing reducing end groups, the commonly used reducing sugar determination method-DNS method is used.

[0044] Pectinase (xylanase) can hydrolyze the substrate pectin (xylan) to generate aldose such as galacturonic acid (xylose), which is produced by co-heating with 3,5-dinitrosalicylic acid Brown-red amino compound, the amount of its reducing sugar is directly proportional to the color depth of the reaction solution containing the colored amino compound, its absorbance is measured at 540nm, and the enzyme activity of pectinase (xylanase) can be calculated.

[0045] (1) DNS reagent

[0046] Dissolve 6.3g of 3,5-dinitrosalicylic acid in 400mL of distilled water, gradually add 21g of sodium hydroxide, then add 185g of potassium sodium tartrate tetrahydrate, 5.0g ...

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Abstract

The invention discloses Bacillus subtilis and application thereof in sisal hemp degumming. The conservation number of the Bacillus subtilis B2 is CCTCCM208115. When in application, the Bacillus subtilis is used as a single strain which is inoculated in seed culture medium, is placed in a gas bath shaker or water bath shaker with temperature of 37 DEG C and is cultured for 8-12h, 2-20 percent (volume percent) of inoculation quantity is trans-inoculated in degumming culture medium when 0D600 reaches 0.6-1, fresh sisal hemp leaves are added in the degumming culture medium, culture temperature is 25-42 DEG C, the revolution of the shaker is 150-250rpm and the sisal hemp degumming can be completed in 35-55h. The invention has the advantages that the culture conditions are extensive, the growth and the reproduction are rapid, the sisal hemp degumming can be completed in 35h, the fiber is not damaged, the environment is not polluted, and the Bacillus subtilis can substitute for mechanical green removal and can be used for the pretreatment of sisal hemp before sisal hemp degumming and pulping technology.

Description

technical field [0001] The present invention relates to the field of microorganisms and microbial fermentation, specifically, the present invention relates to a kind of Bacillus subtilis that can be used for biological degumming or pulping of sisal with the ability to produce pectinase and xylanase, and the bacterial strain can be used in sisal Application in degumming. Background technique [0002] Sisal is a natural fiber raw material with unique advantages in my country. Its fibers have the characteristics of whiteness, toughness, elasticity, and wear resistance. The fiber aspect ratio is large, and the number of interweaving between fibers per unit area is large when it is made into paper. The fiber distribution is fine and the strength of the paper is high. It is an excellent paper-making raw material, especially its high output, fast growth, and short harvesting age, which are inferior to wood and paper-making raw materials. Despite the continuous progress of modern sc...

Claims

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Application Information

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IPC IPC(8): C12N1/20D01C1/04C12R1/125
Inventor 韩双艳林影郑穗平陈青刘志成
Owner SOUTH CHINA UNIV OF TECH
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