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Composite microorganism beta-dextranase and beta-glucosaccharase production method

A technology of glucosidase and compound microorganism, which is applied in the field of production of compound microorganism β-glucanase and β-glucosidase, which can solve the restriction of the application of β-glucanase and the inability to cellobiose and cellotriose Degradation, affecting the hydrolysis efficiency of β-glucan, etc.

Inactive Publication Date: 2008-12-24
天津科建科技发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the absence of β-glucosidase, cellobiose and cellotriose cannot be completely degraded into glucose, which seriously affects the hydrolysis efficiency of β-glucan and restricts the application of β-glucanase

Method used

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  • Composite microorganism beta-dextranase and beta-glucosaccharase production method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Trichoderma viride and Aspergillus niger are mixed and fermented to produce composite microbial β-glucanase and β-glucosidase, made by composite microbial β-glucanase and β-glucosidase Solid enzyme preparation and liquid enzyme preparation, the physical and chemical properties of solid enzyme preparation are:

[0053] (1) Enzyme activity: β-glucanase activity 16000mg glucose / g h, β-glucosidase activity 3300μg glucose / g min,

[0054] (2) Particle size: the preparation can pass through No. 3 standard sieve,

[0055] (3) Moisture: the water content is less than 8%,

[0056] (4) Sensory indicators: yellow powder, uniform color,

[0057] (5) Hygienic indicators: the total number of bacteria per gram of solid enzyme preparation is less than 10,000;

[0058] The physical and chemical properties of liquid enzyme preparations are:

[0059] (1) Product enzyme activity: β-glucanase activity 4500mg glucose / g h, β-glucosidase activity 860μg glucose / g min,

[0060] (2) Sensory i...

Embodiment 2

[0063] Trichoderma viride bacterial classification and Aspergillus niger bacterial classification are made into inclined-plane bacterial classification respectively and preserve;

[0064] Trichoderma viride adopts Chapek`s agar medium (1L) (Czapek`s agar, CZ), and the medium is:

[0065] Sucrose 30g, sodium nitrate 2.0g, dipotassium hydrogen phosphate 1.0g, magnesium sulfate heptahydrate 0.5g, potassium chloride 10.5g, ferrous sulfate 0.01g, agar 15g, pH 7.2-7.4;

[0066] Aspergillus niger adopts potato culture medium (1L) (Potato dextrose agar, PDA), and culture medium is:

[0067] 200g potatoes, 20g glucose, 3g dipotassium hydrogen phosphate, 1.5g magnesium sulfate heptahydrate, 12g agar, natural pH;

[0068] Slant strain culture of Trichoderma viride and Aspergillus niger: use the corresponding culture medium for Trichoderma viride and Aspergillus niger respectively, inoculum size 5-10%, culture statically at 28-30°C for 20-48 hours, put in 4°C refrigerator save as figur...

Embodiment 3

[0070] Inoculate the slant strains of Trichoderma viride and Aspergillus niger respectively in 500ml Erlenmeyer shaker flasks, shake culture on the shaker, mix the medium ratio materials of Trichoderma viride and Aspergillus niger, and mix the culture medium in proportion The materials are divided into conical shake flasks, each bottle is 100 ml, placed in an autoclave, set the temperature at 121 ° C, sterilized for 30 minutes, and after natural cooling, the medium proportioning materials in the conical shake flasks are respectively Inoculate Trichoderma viride strains and Aspergillus niger strains, and carry out shaking flask culture;

[0071] Shake flask culture conditions: temperature 28-30°C, rotation speed 180-250 rpm, inoculum size 1-3%, culture for 48-60h, such as figure 1 shown.

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Abstract

The invention relates to a method for making composite microbio beta-glucanase and beta-glucosidase. The invention includes mixed fermentation of trichoviridin and aspergillus niger to prepare composite microbio beta-glucanase and beta-glucosidase, preparation of solid enzyme preparation and liquid enzyme preparation, and inclined-plane bacterial, shake flask shake culture, liquid seed tank enrichment culture of the trichoviridin and aspergillus niger, mixed input of the trichoviridin and aspergillus niger after enrichment culture into solid fermentation tank to perform fermentation, after fermentation, by digestion, filtration and ultrafilter membrane concentration, obtaining liquid enzyme preparation after adding protecting agent; obtaining solid enzyme preparation by adding British gum in concentrated enzymatic solution with dehumidification and granulation. The inventiom is of scientific and fair industrial design, the trichoviridin and aspergillus niger obtained by mixed culture and screening of strains of fungus can perform paragenetic mixed culture and mutually have no antagonistic reaction, the cost is low, the energy consumption is small, and the invention is of no devil liquor and waste slag discharge, which has a wide applicability of an invention and fills up our zymin breed margin, and has large market foreground.

Description

technical field [0001] The present invention relates to one, in particular to a preparation method of composite microbial β-glucanase and β-glucosidase. Background technique [0002] In recent years, the research direction of β-glucan mainly focuses on the breeding of microbial production bacteria with high β-glucanase activity, and the activity of β-glucanase is used as a reflection indicator of the activity of microbial production bacteria. Ignoring the activity index of β-glucosidase leads to high activity of the produced β-glucanase, but in practical application, the hydrolysis effect is not ideal. The reason is that the β-glucosidase in the enzyme component is very low, even undetectable. However, in the absence of β-glucosidase, cellobiose and cellotriose cannot be completely degraded into glucose, which seriously affects the hydrolysis efficiency of β-glucan and restricts the application of β-glucanase. Contents of the invention [0003] The main purpose of the pr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/24C12N1/14C12N9/98C12R1/685C12R1/885
Inventor 王德培丁友昉杨扬
Owner 天津科建科技发展有限公司
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