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A lipase-producing bacillus and its application

A bacillus and lipase technology, applied in the field of microorganisms, can solve the problems of poor tolerance, easy death, short storage period, etc., and achieve the effects of reducing the material-to-weight ratio, improving economic benefits and protecting the environment

Active Publication Date: 2019-04-05
HUBEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Ordinary bacteria have poor tolerance, short storage period, and are easy to die, so they are not ideal microbial preparations

Method used

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  • A lipase-producing bacillus and its application
  • A lipase-producing bacillus and its application
  • A lipase-producing bacillus and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1: the breeding of bacterial classification

[0018] (1) Media and solutions used in the test

[0019] Each 1000g enrichment medium contains: waste oil 10mL, yeast extract 2g, NaCl 0.5g, K 2 HPO 4 1g, MgSO 4 · 7H 2 O 0.1g, (NH 4 ) 2 SO 4 1g, the balance is water, the pH is natural;

[0020] Each 1000g of selective separation medium contains: 10g of tryptone, 5g of yeast extract, 10g of NaCl, 2mL of tributyrin, 20g of agar, the balance is water, and the pH is natural;

[0021] Each 1000g shake flask seed medium contains: glucose 20g, K 2 HPO 4 1g, MgSO 4 · 7H 2 O 0.5g, (NH 4 ) 2 SO 4 5g, 25g of peptone, 10mL of waste oil, the balance is water, and the pH is natural;

[0022] Each 1000g solid slant medium contains: tryptone 10g, yeast extract 5g, NaCl 10g, agar 20g, the balance is water, and the pH is natural;

[0023] The re-screened fermentation medium on every 1000g shake flask contains: glucose 5g, K 2 HPO 4 1g, MgSO 4 · 7H 2 O0.5...

Embodiment 2

[0028] Example 2: Bacillus Bacillus Optimization of culture conditions for sp. HFE722

[0029] (1) Bacterial strain: the strain bacillus that embodiment 1 screens out Bacillus sp. HFE722.

[0030] (2) Method steps:

[0031] single factor test Bacillus Effects of sp. HFE722 fermentation temperature, initial pH, fermentation cycle, inoculum volume, and liquid volume on lipase production.

[0032] The fermentation medium was the initial medium (glucose 5g, K 2 HPO 4 1g, MgSO 4 · 7H 2 O 0.5g, (NH 4 ) 2 SO 4 1g, yeast powder 20g, waste oil 10mL, the balance is water). The initial fermentation conditions were: natural pH, inoculum size 1% (v / v), liquid volume 100 mL / 250 mL, 30°C, 160r / min shake flask fermentation for 36h.

[0033] a. Optimal test of fermentation cycle

[0034] Other fermentation conditions are the initial fermentation conditions, and samples are taken every 6 hours to measure the lipase enzyme activity, and the determination of Bacillus Bacillus Opti...

Embodiment 3

[0043] Embodiment 3: Effect of waste oil, olive oil, soybean oil, peanut oil, cottonseed oil on strain HFE722 producing enzyme

[0044] The gutter oil in the initial medium was replaced with olive oil, soybean oil, peanut oil and cottonseed oil, and the effects of various oil additives on the enzyme production of the strain were investigated under the same addition amount.

[0045] Figure 6 The control group refers to no oil added. It can be seen from the figure that the effects of waste oil, olive oil and cottonseed oil on the enzyme activity are not much different. When the additive is waste oil, the enzyme activity is up to 6.3U / mL. From the perspective of cost and environmental protection, waste oil can perfectly replace other oil species as the enzyme production promoter of the medium.

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Abstract

The invention discloses a bacillus Bacillus sp.HFE722 and its application, this strain has been preserved in China Center for Type Culture Collection on December 10, 2015, and the preservation number is: CCTCC NO:M 2015735. The bacterial strain bred by the present invention can produce high-yield lipase, and waste oil is used as a raw material, so that the waste can be reused, the environment is protected, and sustainable development is realized; the bacterial strain can significantly improve the feed conversion rate, improve the growth performance of livestock as a feed additive, Regulating the intestinal microflora and reducing the feed-to-weight ratio provide an important theoretical and practical basis for reducing the cost of livestock and poultry farming and improving economic benefits.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, in particular, relates to a bacillus with strong ability to produce lipase ( Bacillus sp . ) and its applications. Background technique [0002] Lipase is a kind of special ester bond hydrolase, and it is one of the most in-demand biocatalysts, generally used to catalyze hydrolysis and synthesis reactions. The catalytic activity of lipase mainly lies in its protein structure. At the oil-water interface, it catalyzes the hydrolysis of the ester bond of triacylglycerol to generate monoglyceride, diglyceride or directly generate glycerol and fatty acid. [0003] Lipases of microbial origin are most abundant, including those of bacterial, mold, and yeast origin. Due to the variety of microorganisms and rapid reproduction, the lipase produced has a wider pH value than animal and plant lipases, and the lipases derived from microorganisms are generally secreted extracellular enzymes, which are...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/20A23K20/189
CPCC12N9/20C12Y301/01003C12N1/205C12R2001/07
Inventor 蔡俊胡珺王常高杜馨林建国
Owner HUBEI UNIV OF TECH