Enterobacter ludwigii and application thereof

A technology of Enterobacter reuteri and MY271, applied in the field of microbiology, can solve the problems of unreported β-mannanase, etc., and achieve the effects of lowering cholesterol levels, improving biological regulation functions, and promoting growth

Inactive Publication Date: 2015-08-05
HUBEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] After reviewing the existing technologies at home and abroad, there is no report on the use of Enterobacter reuteri to produce β-mannanase using konjac flour

Method used

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  • Enterobacter ludwigii and application thereof
  • Enterobacter ludwigii and application thereof
  • Enterobacter ludwigii and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0033] The preparation method of the crude enzyme liquid is as follows: take a certain volume of bacterial liquid in a centrifuge tube, centrifuge at 8000rpm for 15 minutes to get the supernatant, and dilute it by an appropriate multiple to obtain the crude enzyme liquid.

[0034] The method for measuring the enzyme activity of β-mannanase is as follows: take 1.5mL of 0.5% konjac purified powder solution in a 25mL colorimetric tube, add 0.5mL of crude enzyme solution and react in a water bath at 55°C for 10min, add 3mL of DNS solution, and place in a boiling water bath After boiling in medium for 5 minutes, cool to room temperature immediately, add distilled water to make up to 25mL, this is the experimental group; the control group uses the boiled inactivated enzyme solution instead of the crude enzyme solution. Absorbance was measured at 540 nm.

Embodiment 1

[0035] Example 1: Optimization of culture conditions for Enterobacter reuteri MY271

[0036] (1) Strain: Enterobacter reuteri MY271CCTCC M 2015182

[0037](2) Method steps:

[0038] The fermentation medium was the initial medium (konjac flour 5.0g / L, peptone 5.0g / L, KH 2 PO 4 1.0g / L, MgSO 4 0.1g / L). The initial fermentation conditions are: natural pH, 5% inoculum size, 50mL / 250mL liquid volume, 30°C, 160r / min shake flask fermentation for 48h.

[0039] The single factor experiment was used to investigate the effects of the inoculum amount, liquid volume, initial pH, fermentation temperature and fermentation cycle of Enterobacter reuteri MY271 strain on the production of β-mannanase.

[0040] a. Optimization test of inoculum size

[0041] Other fermentation conditions are the initial fermentation conditions. When the inoculum size is 3%, 5%, 7%, 9%, 11%, 13% and 15%, the effect of the inoculum size on the production of β-mannanase by Enterobacter reuteri MY271 is investi...

Embodiment 2

[0055] Example 2: Optimization of medium components for Enterobacter reuteri MY271

[0056] (1) Bacterial strain: the bacterial species Enterobacter reuteri MY271 screened out in Example 1

[0057] (2) Method steps:

[0058] The culture conditions are fermentation time 4h, temperature 31°C, inoculum size 9%, initial pH 7.0, liquid volume 50mL / 250mL.

[0059] a. Carbon source selection and addition test

[0060] Konjac powder, locust bean gum, sucrose, D-mannose, D-mannitol, soluble starch, maltose, glucose, D-fructose, α-lactose, sodium carboxymethylcellulose, D-xylose, molasses as carbon Source variable, the addition amount is 5g / L, and other components are initial medium components.

[0061] Depend on Figure 6 It can be seen that when konjac flour was used as the carbon source, strain MY271 had the best effect of inducing enzyme production, and the enzyme activity was 8.01 U / mL, followed by locust bean gum. Therefore, konjac flour was selected as the best carbon source....

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PUM

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Abstract

The invention discloses enterobacter ludwigii and application thereof. A bacterial strain is enterobacter ludwigii MY271 collected in China Center for Type Culture Collection (CCTCC) in March 31, 2015, and has a collection number of CCTCC M2015182. According to the bred bacterial strain disclosed by the invention, beta-mannanase can be produced with high yield by using konjaku flour, manno-oligosaccharides can also be obtained, and the obtained manno-oligosaccharides have very good biological regulation functions, can be used for effectively reducing the cholesterol level of a human body, reducing blood sugar and promoting the growth of bifidobacteria in intestinal tracts, and are good food additives.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to an Enterobacter ludwigii with strong ability to produce β-mannanase by using konjac flour and its application. Background technique [0002] Hemicellulose is the second largest class of heteropolysaccharides in nature, of which β-mannan is a major hemicellulose. β-Mannan widely exists in locust bean gum, guar gum, konjac powder and other plant polysaccharides. β-mannanase (β-mannanase, EC 3.2.1.78) can hydrolyze polysaccharides containing β-1,4 mannosidic bonds on the main chain (β-mannan, glucomannan, galactomannan Sugar) is mannooligosaccharide. The obtained manno-oligosaccharide has a good biological regulation function, can effectively reduce the cholesterol level of the human body, lower blood sugar, and promote the growth of bifidobacteria in the intestinal tract, and is a good food additive. The enzyme is also widely used in many aspects such as feed, paper...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/42C12P19/14C12R1/01
Inventor 蔡俊杨苗王常高杜馨林建国
Owner HUBEI UNIV OF TECH
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