Thermophilic acid-resisting beta-glucosaccharase producing strain and preparation method thereof

A glucosidase and acid-resistant technology, which is applied in the fields of thermophilic acid-resistant β-glucosidase producing bacteria and preparation, can solve the problems of high price of β-D-glucoside hydrolase and no way to realize industrialized use and the like

Inactive Publication Date: 2010-08-04
JILIN UNIV
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Problems solved by technology

Simultaneously, due to the limitation of production strain and production conditions, the price of commercialized β-D-glucoside hydrolase is very high, there is no way to realize large-scale industrialized use, and the task of the present invention is to find a kind of β-D-glucoside hydrolase that can be produced with cheaper raw materials Excellent strain of thermophilic and acid-resistant β-glucosidase

Method used

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  • Thermophilic acid-resisting beta-glucosaccharase producing strain and preparation method thereof
  • Thermophilic acid-resisting beta-glucosaccharase producing strain and preparation method thereof
  • Thermophilic acid-resisting beta-glucosaccharase producing strain and preparation method thereof

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Embodiment 1

[0026] Strain screening method

[0027] Collect the rotten parts of corn stalks that have accumulated for many years, cut them into small pieces with scissors, break them with glass beads in 100mL sterilized water, and insert the obtained bacterial liquid into the medium containing only cellobiose as carbon source for 50 days. ℃ enrichment culture, the enrichment medium is: peptone 2.5g / L, cellobiose 2.5g / L, (NH 4 ) 2 SO 4 1g / L, KH 2 PO 4 0.5g / L, MgSO 4 0.2g / L. The culture conditions are: 50°C, 120rpm, enrichment culture for 3~5d; -1~10 -6 Take 100μL and smear it on a coarse sieve plate. The coarse sieve medium is: yeast powder 2.5g / L, peptone 2.5g / L, cellobiose 0.25g / L, (NH 4 ) 2 SO 4 1g / L, KH 2 PO 4 0.5g / L, MgSO 4 0.2g / L, agar 15~20g / L, add filtered 1g escin and 2.5g ferric ammonium citrate solution after sterilization, culture at 30℃ for 24h, put the plate in the refrigerator for 1h to develop color, observe the transparent circle In case of occurrence, select...

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Abstract

The invention discloses a thermophilic acid-resisting beta-glucosaccharase producing strain and a preparation method thereof. The strain (CCTCC M209312) is separated from decayed corn stalks. The screening method comprises the following steps: collecting corn stalks suitable for decaying, cutting, diluting with sterilized water, carrying out induction enrichment culture with cellobiose, carrying out gradient concentration dilution, coating on a screening plate, selecting the colony with larger plaques as the primary screening, and carrying out primary screening for times; and after culturing with a steam-explosion corn stalk powder induction culture medium, measuring the enzyme activity and carrying out second screening, and centrifugating the screened strain fermentation liquid to obtain the supernatant. The supernatant can be used as a liquid enzyme preparation or spray-dried to obtain a solid enzyme preparation, and can adapt to various acid high-heat industrial conditions. The invention screens out a new strain from decayed corn stalks by using biotechnology, and uses steam-explosion stalk powder and other cheap culture media to produce the thermophilic acid-resisting beta-glucosaccharase. The invention is used in the industrial fields of cellulose degradation systems, food, textile, papermaking and the like.

Description

Technical field: [0001] The present invention relates to a thermophilic acid-resistant β-glucosidase producing bacterium and a preparation method thereof, and specifically provides a method for screening and fermenting a thermophilic acid-resistant β-glucosidase producing bacterium and utilizing the produced bacterium The practical application of enzymes in industrial processes belongs to the related fields of microbial fermentation and its product utilization. Background technique: [0002] β-glucosidase (EC3.2.1.21), also known as β-D-glucoside hydrolase, can hydrolyze the non-reducing β-D-glucosidic bond bound to the terminal and release β-D-glucose at the same time and the corresponding ligands. It widely exists in many plants, insects, yeasts, Aspergillus, Trichoderma and bacteria in nature. It participates in the glucose metabolism of organisms and plays an important role in maintaining the normal physiological functions of organisms. β-glucosidase is also very impo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/42C12R1/645
Inventor 滕利荣林凤张益波孙树祯孟庆繁于占春逯家辉张小希田鸿儒邓红波刘海龙张盈刘青毛国涛张晗思
Owner JILIN UNIV
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