Method of producing neutral beta-konjak mannase preparation
A technology of mannanase and production method, which is applied in the field of microbial fermentation technology and enzyme engineering, can solve the problems of complex process, high cost, unfavorable environmental protection, etc., and achieve the effects of increasing dissolved oxygen, simple method, and reducing viscosity
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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, 0.5% NaCl, 0.3% Congo red; 30 ~ 35 ℃, culture 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 co...
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, P...
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