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Acid-resistant alpha-amylase

An amylase and enzyme gene technology, applied in the field of acid-resistant α-amylase, can solve the problems of increasing environmental burden and complex processing technology, and achieve the effects of simplifying starch saccharification process, reducing environmental pollution, and high enzyme activity

Inactive Publication Date: 2021-07-23
广州博识生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In the sugar refining and fermentation industry, starch is the main raw material. In the starch processing process, the starch milk is liquefied to saccharified, and the pH is 4.5, 6.5, and 4.5 respectively. During this period, acid and alkali need to be added to adjust the pH, and acid is added during the process. Alkali will complicate the processing technology and increase the burden on the environment. If the α-amylase used for liquefaction has strong acid stability, there is no need to adjust the pH. Therefore, it is necessary to research and develop acid-resistant α-amylase

Method used

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Experimental program
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Embodiment 1

[0028] The materials and reagents involved in this embodiment are shown in Table 1, and the experimental instruments are shown in Table 2;

[0029] Table 1 Experimental materials and reagents

[0030]

[0031]

[0032] Table 2 Experimental equipment

[0033] Gene shock introduction instrument Bio-Rad Bole T100 PCR instrument Beijing Century Kexin Scientific Instrument Co., Ltd. BBS-H1100 clean bench Shandong Boke Biological Industry Co., Ltd. DYY-6D Nucleic Acid Electrophoresis Instrument Beijing Liuyi Instrument Factory Mini Protein 3 protein electrophoresis system American Bio-Rad Corporation GelDoc gel imaging system American Bio-Rad Corporation UV1900 UV Visible Spectrophotometer Ranch (Shanghai) Instrument Technology Co., Ltd. TGL-16MB high speed refrigerated centrifuge Changsha Xiangzhi Centrifuge Instrument Co., Ltd. THZ-300 constant temperature culture shaker Shanghai Yitian Scientific Instrument ...

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Abstract

The invention relates to acid-resistant alpha-amylase. An alpha-amylase gene sequence (alpha-amylas e, GenBank: KX216807.1) is obtained from National Center of Biotechnology Information (NCBI); a mutation product is obtained through error-prone polymerase chain reaction(PCR); the mutation product is recombined into pichia pastoris to be expressed, mutons with high specific enzyme activity under environments of pH3.5, pH4.5, pH5.5, pH6.5 and pH7.5 are screened to obtain an acid-resistant muton, and a gene sequence of the acid-resistant muton is mutated into TT from original CC at a 430 site and a 431 site. The invention has the beneficial effects that the obtained mutant alpha-amylase still has high enzyme activity in the environment of pH4.5; a starch saccharification process can be simplified, and environmental pollution is reduced.

Description

technical field [0001] The invention belongs to the field of genetic engineering and enzyme engineering, in particular to an acid-resistant alpha-amylase. Background technique [0002] α-amylase, α-1,4-glucan-4-glucan hydrolase (EC.3.2.1.1), α-amylase randomly acts on starch, glycogen, oligosaccharide or polysaccharide molecules The α-1,4-glucosidic bond in the starch is hydrolyzed into small molecular substances such as glucose, maltose, and oligosaccharides. Amylase is a class of enzyme products with the widest application and the largest output. Industrial α-amylases are divided into two categories: bacterial α-amylases and fungal α-amylases according to different microbial sources. The main products of bacterial α-amylase hydrolyzing starch are dextrin and a small amount of oligosaccharides and glucose. Fungal α-amylase can hydrolyze starch and maltotriose, and the final products are mainly maltose, some oligosaccharides and a small amount of glucose. Due to its excel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/26C12N15/81C12N15/56
CPCC12N9/2414C12N15/815C12Y302/01001C12N2800/102
Inventor 刘庚敬
Owner 广州博识生物科技有限公司