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Preparation method of heat-resistant beta-galactosidase

A galactosidase, heat-resistant technology, applied in the field of genetic engineering and enzyme engineering, can solve the problems of easy contamination of bacteria, low unit yield of lactase, poor heat resistance, etc.

Inactive Publication Date: 2021-04-13
马金佑
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] A lot of research work has been done on lactase at home and abroad, but it has not been widely used. The main reasons are: 1. The unit yield of lactase is low; Obtained, the extraction and purification process is complicated, and the cost is high; 3. The action temperature of the enzyme is low, the heat resistance is poor, and it is easy to be contaminated with bacteria in industrial production, and the scope of application is narrow

Method used

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  • Preparation method of heat-resistant beta-galactosidase
  • Preparation method of heat-resistant beta-galactosidase
  • Preparation method of heat-resistant beta-galactosidase

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Embodiment 1

[0035] The materials and reagents involved in this embodiment are shown in Table 1, and the experimental instruments are shown in Table 2;

[0036] Table 1 Experimental materials and reagents

[0037] E.coli T-A cloning vector pET20b(+) Shanghai Lianmai Bioengineering Co., Ltd. Plasmid Miniprep Kit Tiangen Biochemical Technology Co., Ltd. Ampicillin Shanghai Tongwei Biological Co., Ltd. Coomassie Brilliant Blue R-250 Beijing Solaibao Technology Co., Ltd. Agarose Gel DNA Recovery Kit Dalian Bao Biological Engineering Co., Ltd. restriction endonuclease Dalian Bao Biological Engineering Co., Ltd. Taq enzyme Dalian Bao Biological Engineering Co., Ltd. In-Fusion Cloning Kit Dalian Baosheng Biological Engineering Co., Ltd. yeast Dxoid Ltd. England tryptone Dxoid Ltd. England Nitrogen source for yeast NEB Pichia pastoris GS115 Invitrogen o-Nitrophenol-β-D-galactoside Applichem Corporation ...

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Abstract

The invention relates to a preparation method of heat-resistant beta-galactosidase. The preparation method of the heat-resistant beta-galactosidase is characterized in that an enzyme gene sequence (beta-galactosidase, GenBank: NT_166518.1) is mutated on the 2010th site and mutated from the original A to C. The preparation method has the beneficial effect that the optimal reaction temperature of the obtained mutated beta-galactosidase is greatly increased.

Description

technical field [0001] The invention belongs to the field of genetic engineering and enzyme engineering, in particular to the preparation of heat-resistant beta-galactosidase. Background technique [0002] Lactose intolerance is a disease of lactose indigestion. The main reason is the lack of lactase in the body or its low activity. Lactose cannot be hydrolyzed into simple sugars by lactase, and the human body cannot absorb it. The incidence of lactose malabsorption in adults in my country is as high as 86.7%. [0003] Lactase, also known as β-D-galactoside galactohydrolase, can hydrolyze lactose into glucose and galactose. Lactase is widely used in medicine, dairy products, immunoassay detection, chemical synthesis, biology, There are applications in the field of environmental protection. At present, it is recognized that the safe strains for preparing lactase are mainly probiotics, Kluyveromyces lactis, Aspergillus oryzae, Aspergillus niger, and Kluyveromyces fragilis. ...

Claims

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Application Information

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IPC IPC(8): C12N9/38C12N15/56C12N15/70
CPCC12N9/2471C12N15/70C12Y302/01023
Inventor 马金佑
Owner 马金佑
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