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Bacterial strain for producing beta-mannanase and production process of bacterial strain

A technology of mannanase and bacterial strains, applied in the field of bioengineering, can solve the problems of high cost, low activity of mannanase, and low yield, and achieve good pH value and temperature stability, good stability, and high enzyme activity Effect

Active Publication Date: 2011-08-10
浙江皇冠科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Secondly, the mannanase activity produced by the screened strains is low, and the substrate-induced fermentation process is complicated, the cost is high, and the yield is low.

Method used

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  • Bacterial strain for producing beta-mannanase and production process of bacterial strain
  • Bacterial strain for producing beta-mannanase and production process of bacterial strain
  • Bacterial strain for producing beta-mannanase and production process of bacterial strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Bacterial Strain Isolation and Purification

[0021] 1. Medium

[0022] Enrichment medium (g / L): konjac powder 20, KH 2 PO 4 0.5, pH natural;

[0023] Separation medium (g / L): konjac powder 5.0, yeast extract 2.0, peptone 10, KH 2 PO 4 0.1, MgSO 4 ·6H 2 O 0.12, agar 20.0, pH natural;

[0024] Fermentation medium (g / L): konjac powder 20.0, yeast extract 5.0, NaNO 3 5.0, MgSO 4 ·6H 2 O0.2,K 2 HPO 4 5.0, pH natural;

[0025] 2. Isolation of strains

[0026] Collect the soil samples from the konjac planting ground, dilute them with sterile water and let them stand, take an appropriate amount of supernatant and inoculate them into the enrichment medium, cultivate them on a shaker at 37°C for 36 hours, and take 1.0mL of the culture solution with the most obvious viscosity drop in the shaker flask , after gradient dilution according to the conventional method, spread on the separation medium plate, and then put it in a constant temperature incubator for...

Embodiment 2

[0027] The preliminary identification of embodiment 2 bacterial strains

[0028] According to the "Common Bacteria System Identification Handbook" and "Microbiology Experiment Handbook", the preliminary identification of its morphological characteristics, culture characteristics, physiological and biochemical assays, etc. was carried out.

[0029] The strain is a Gram-positive bacillus with a capsule. It can grow in the temperature range of 20-60°C, the optimum growth temperature is 37-50°C, the growth pH is 5-11, and the optimum growth pH is 7-8. Ability to hydrolyze starch, able to withstand 7% NaCl growth, nitrate reduction, catalase determination, V.P experiment, citrate and propionate utilization, nitrite reduction, gelatin liquefaction, methyl red experiment, casein hydrolysis results It was positive, and the results of hydrogen sulfide test and indole test were negative. The strain was identified and named as Bacillus subtilis (Bacillus subtilis) Bacillus subtilis-hzb...

Embodiment 3

[0030] Example 3 Fermentative production of β-mannanase

[0031] Inoculate the bacterial strain into the seed medium and cultivate it for 24 hours to obtain the seed liquid, put the seed liquid into a 250ml Erlenmeyer flask containing 30ml of fermentation medium, and place it in a rotary constant temperature and speed regulating shaker cabinet for aerobic fermentation , rotate at 200r / min, ferment at 37°C for 24 hours, centrifuge the fermentation broth at 1000r / min for 10 minutes, take the supernatant, which is the crude enzyme solution of β-mannanase, and store at 4°C.

[0032] Seed medium (g / L): sodium chloride 10, peptone 10, yeast extract 5, agar 20.

[0033] Fermentation medium (g / L): konjac powder 20.0, yeast extract 5.0, K 2HPO 4 5.0g, MgCl 2 .·6H 2 O 0.2, NaNO 3 5.0, pH5.5.

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Abstract

The invention discloses a bacterial strain for producing beta-mannanase and a production process of the bacterial strain. The bacterial strain is named as bacillus subtilis-hzbzbc and preserved in the China General Microbiological Culture Collection Center (CGMCC) located at No.3 building, No. 1 courtyard, Beichen West Road, Chaoyang District, Beijing, the preservation number is CGMCC No. 4358, and the preservation time is November 22, 2010. The activity of the beta-mannanase produced by the bacterial strain is high, the optimum pH value is 2.5, the optimum temperature is 50 DEG C, and the beta-mannanase has good pH value and temperature stability. Meanwhile, the stability of the mannanase to metal ions and chelating agents is good.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a bacterial strain for producing β-mannanase and a production process. Background technique [0002] β-mannanase (β-mannanase; endo-1,4-β-D-mannanmannohydrolase; EC3.2.1.78, referred to as mannanase) is the main hydrolysis of 1,4-β-D-mannanmannose The chain endohydrolase belongs to hemicellulase, and the main substrates are galactomannan, glucomannan, galactoglucomannan and mannan. [0003] Mannanase exists widely in nature, and enzyme activity has been found in some lower animals, such as marine mollusk intestinal secretions, germinated seeds of some legume plants, and germinated bulbs of Araceae plant Amorphophallus konjac The presence. For microorganisms, it is the main source of producing mannanase. It has been reported that Bacillus, Pseudomonas, and Vibrio in bacteria, Aspergillus, Trichoderma, and yeast in fungi, and Streptomyces in actinomycetes are common group...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N9/42C12R1/125
Inventor 胡向东胡佳佳胡伟卿
Owner 浙江皇冠科技有限公司
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