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Method of producing neutral beta-konjak mannase preparation

A konjac and neutral technology, applied in the field of microbial fermentation technology and enzyme engineering, can solve the problems of high cost, complex process, unfavorable environmental protection, etc., and achieve the effect of reducing viscosity, increasing dissolved oxygen, and simple method

Inactive Publication Date: 2007-08-15
WUHAN TAIYUAN INVESTMENT GUARANTEE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In some of the above studies, either the enzyme activity is not high, the enzyme production is unstable, or the process is complex, the product conversion rate is low, and the cost is high
If it is produced with alkaline mannanase, the enzymatic hydrolysis process is carried out under the condition of PH9-10, which brings difficulties to the post-processing, affects the taste of the product, and is not conducive to environmental protection

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Production of High Vitality Neutral β-Konjac Mannanase Strain Breeding

[0027] 1) Based on the 10164 strain screened by CICC, through UV mutagenesis, the strains were planted in 0.3% Congo red plate medium, and the medium formula consisted of 2.0% agar, 1% konjac powder, pancreatic 0.5% peptone, 0.5% yeast extract powder, 10.5% NaC, 0.3% Congo red; 30-35°C, cultured for 35 hours, a transparent hydrolysis circle can be seen.

[0028] 2) Select the bacterial strain with the larger H / C colony diameter value of the hydrolysis circle to carry out shake flask re-screening, and the shake flask medium is composed of: konjac degradation solution (brix 10BX) 500ml, tryptone 30g, yeast extract powder 50g, Na 2 HPO 4 12H 2 O 100g, KH 2 PO 4 10g, NaCl 20g, NH 4 Cl 10g, soybean meal 200g, corn dregs 100g, pH 6.0, cultured at 35°C for 30h to obtain crude enzyme liquid, and the enzyme activity was determined by DNS method. Select excellent strains for stable passage f...

Embodiment 2

[0029] Example 2 Fermentation production of high-activity neutral β-konjac mannanase by strain TQS6 CCTCC M207001

[0030]1) First-level liquid culture: scale-up culture from inclined planes to triangular flasks, the composition of the medium is: 150ml of konjac degradation solution, 10g of yeast extract powder, 10g of tryptone, 5g of NaCl, the rest is water, add water to 1000ml, PH6.5~7.5 , the inoculum amount is one loop of slant bacteria, cultivated in liquid medium for 12-24 hours at 30-35°C, 200-300r / min, and pH6.5-7.5.

[0031] 2) Enzyme production by small tank fermentation: 10L fermenter, medium composition konjac degradation solution (sugar content 10BX) 500ml, tryptone 100g, yeast extract powder 100g, Na 2 HPO 4 12H 2 O 100g, KH 2 PO 4 20g, NaCl25g, NH 4 Cl 5g, soybean meal powder 250g, corn dregs 100g, the rest is water, add water to 5 liters. Inoculate 200ml of first-grade liquid seeds, and culture continuously for 24-48 hours at 30-35°C, 200-500r / min, DO con...

Embodiment 3

[0036] Example 3 Production of high-activity neutral β-konjac mannanase by high-density cell culture and fermentation

[0037] 1) First-level liquid culture: scale-up culture from inclined plane to triangular flask, medium composition: 150ml of konjac degradation solution, 10g of yeast extract powder, 10g of tryptone, 5g of NaCl, the rest is water, add water to 500ml, PH6.5~7.5 , the inoculum amount is one loop of slant bacteria, cultivated in liquid medium for 12-24 hours at 30-35°C, 200-300r / min, and pH6.5-7.5.

[0038] 2) Enzyme production by small tank fermentation: 10L fermenter, medium composition konjac degradation solution (sugar content 10BX) 1000ml, tryptone 100g, yeast extract powder 100g, Na 2 HPO 4 12H 2 O 100g, KH 2 PO 4 25g, NaCl 25g, NH 4 Cl 5g, soybean meal 250g, corn dregs 100g, perfluorocarbon 50g, Tween 80 50g, the rest is water, add water to 5 liters. Inoculate 400ml of first-grade liquid seeds, and culture continuously for 24-48 hours at 30-35°C, PH...

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PUM

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Abstract

The invention discloses a preparing method of bacillus subtilis TQS6 CCTCC M207001 of highly effective expression neutral beta-mannase through adopting UV (ultraviolet mutagenesis ) and NTG (nitrosoguanidine mutagenesis) compositing mutagenesis, which comprises the following steps: bevel-culturing bacterial strain; proceeding ferment culture after one grade and two grade culture; choosing culture medium; adding into not exclusively degraded liquid from centrifuging sugar liquid; venting into oxygen-enriched air during the course of yeast; increasing dissolved oxygen; getting high active neutral beta-konjak mannase. The product possesses high enzyme production, high enzyme activator, short period of production and low cost, which can invert low mannan effectively.

Description

technical field [0001] The invention relates to microbial fermentation technology and enzyme engineering technology, in particular to a production method of bacillus subtilis CCTCCM207001 and its high-activity neutral β-konjac mannan enzyme preparation. Background technique [0002] Mannan saccharification is a class of endohydrolase that can hydrolyze mannan polysaccharides containing mannosidic bonds, and it belongs to the class of hemicellulase. Mannan polysaccharides mainly exist in konjac, guar gum, and selenium gum, mainly including glucomannan, galactomannan, galactoglucomannan, etc., which constitute the second largest group of plant hemicelluloses The enzymes that mainly depend on the degradation of these substances are mannanases. Since the 1990s, the research on mannanase and its enzymatic preparation of oligosaccharides has attracted great attention. Ma Yanhe, Institute of Microbiology, Chinese Academy of Sciences, etc. studied the production conditions of alka...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/24C12N13/00
Inventor 彭冬秋黄代勇刘全新万霞李小虎黄芳
Owner WUHAN TAIYUAN INVESTMENT GUARANTEE
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