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Method for producing low temperature-resistance beta-mannase by using enterobacter

A mannanase and low temperature-resistant technology, which is applied in the field of producing β-mannanase and achieves the effects of high enzyme activity, simple cultivation, wide temperature and pH range

Inactive Publication Date: 2015-04-22
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no report on the production of β-mannanase by Enterobacteriaceae

Method used

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  • Method for producing low temperature-resistance beta-mannase by using enterobacter
  • Method for producing low temperature-resistance beta-mannase by using enterobacter
  • Method for producing low temperature-resistance beta-mannase by using enterobacter

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Fermentation medium (wt%): guar gum 1.0%, NH 4 NO 3 0.1%, KH 2 PO 4 0.245wt%, Na 2HPO 4 12H 2 O 1.0%, MgSO 4 0.01%, yeast powder 0.05%, trace element 10mL (trace element (wt%): MnSO 4 ·H 2 O 0.03%, Na 2 MoO 4 2H 2 O 0.01%, FeSO 4 ·7H 2 O 0.2%, CuSO 4 0.006%, CaCl 2 0.1%). Enterobacter sp.N18 was cultivated in shake flasks at 37° C. at 150 r / min, with an inoculum size of 5%, and cultivated for 48 hours. Using guar gum as a substrate at 50°C, the enzyme activity detected by DNS method was 596.7U / mL; within 24 hours, the viscosity of 1.4% guar gum solution was reduced from 800mPa·s to 78mPa·s, a reduction of 90%.

Embodiment 2

[0031] Fermentation medium (wt%): carbon source 1.0%, NH 4 NO 3 0.1%, KH 2 PO 4 0.245wt%, Na 2 HPO 4 12H 2 O 1.0%, MgSO 4 0.01%, yeast powder 0.05%, trace element 10mL (trace element (wt%): MnSO 4 ·H 2 O 0.03%, Na 2 MoO 4 2H 2 O 0.01%, FeSO 4 ·7H 2 O 0.2%, CuSO 4 0.006%, CaCl 2 0.1%). The carbon sources are guar gum, konjac flour, locust bean gum, D-mannan, D-galactose, maltose, glucose, sucrose, soluble starch, and dextrin. Enterobacteriaceae were cultured in shake flasks at 37°C and 150r / min (Enterobacter sp.N18), the inoculum size was 5%, and cultured for 48 hours. Using guar gum as a substrate at 50°C, the enzyme activity was determined by the DNS method, and the results are shown in Table 1. figure 1 .

[0032] Table 1: Detection results of β-mannanase activity obtained from different carbon sources

[0033] carbon source

[0034] Enzyme activity (U)

[0035] .

Embodiment 3

[0037] Fermentation medium (wt%): konjac flour 0.1~3.0%, NH 4 NO 3 0.1%, KH 2 PO 4 0.245wt%, Na 2 HPO 4 12H 2 O 1.0%, M g SO 4 0.01%, yeast powder 0.05%, trace elements 10 mL (trace elements (wt%): MnSO 4 ·H 2 O 0.03%, Na 2 MoO 4 2H 2 O 0.01%, FeSO 4 ·7H 2 O 0.2%, CuSO 4 0.006%, CaCl 2 0.1%). Enterobacter sp.N18 was cultivated in shake flasks at 37° C. at 150 r / min, with an inoculum size of 5%, and cultivated for 48 hours. Using guar gum as a substrate at 50°C, the enzyme activity was detected by the DNS method, and the results are shown in Table 2.

[0038] Table 2: The enzyme activity detection results of β-mannanase cultured with different concentrations of konjac flour as carbon source

[0039] Konjac flour (wt%)

[0040] .

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Abstract

The invention relates to a method for producing low temperature-resistance beta-mannase by using enterobacter. The enterobacter sp. N18 is inoculated to a medium containing carbon source and nitrogen source, and is cultured for 12-96 hours at 37 DEG C, wherein a broth is a beta-mannanase product. Compared with the prior art, the natural enterobacter can be naturally screened for fermentation and culture to obtain the beta-mannanase, cost is low, and operation is simple. The beta-mannanase has the advantages of high enzyme activity and good stability, activity can be kept under condition that temperature is 0-100 DEG C, pH value is 3.0-11.0. and salinity is in 0-4.5M scope, and specific activity can reach 5484U / mg at 20 DEG C. In addition, viscosity of a guar gum solution with viscosity of more than 800 mPa.s can reduced to more than 95% in 10 minutes by the enzyme.

Description

technical field [0001] The invention relates to a method for producing beta-mannanase, in particular to a method for producing low-temperature-resistant beta-mannanase by fermenting and culturing enterobacteria. Background technique [0002] Hemicellulose is the second largest resource on the earth, and mannan, as a main component of hemicellulose, has great development value. Mannanase is needed for the development and utilization of mannan resources. With the development of mannan resources, β-mannanase has a wide development space in food, medicine, papermaking, feed, textile printing and dyeing, oil exploration and other industrial fields. [0003] β-mannanase (β-1,4-D-mananmannohydrolase; EC 3.2.1.78) can hydrolyze mannan oligosaccharides and mannan polysaccharides (including mannan, glucose mannan and galactomannan, etc.). β-mannanase comes from a wide range of sources, including animals, plants, bacteria, actinomycetes, and fungi, among which microorganisms are the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12R1/01
CPCC12N9/2494C12Y302/01078
Inventor 牟伯中杨世忠刘金峰刚洪泽尤佳
Owner EAST CHINA UNIV OF SCI & TECH
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