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A modified β-glucosidase and its expression gene and application

A technology for glucosidase and gene expression, which is applied to the modified beta-glucosidase and its expression gene and application fields, and can solve the problems of strict reaction conditions, harsh substrate concentration requirements, and difficulty in precise control of reaction time.

Active Publication Date: 2017-12-05
SHANDONG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method has problems such as strict requirements on the substrate concentration, strict requirements on reaction conditions, and difficulty in accurately controlling the reaction time.
So far, there is no report on the simultaneous production of laminaribiose, cellobiose and sophorose by enzymatic method

Method used

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  • A modified β-glucosidase and its expression gene and application
  • A modified β-glucosidase and its expression gene and application
  • A modified β-glucosidase and its expression gene and application

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Embodiment Construction

[0007] Technical scheme of the present invention is as follows:

[0008] A modified β-glucosidase, the amino acid sequence of which is shown in SEQ ID NO.1.

[0009] A modified expression gene of β-glucosidase, the nucleotide sequence of which is shown in SEQ ID NO.2.

[0010] A recombinant expression vector obtained by inserting the nucleotide sequence shown in SEQ ID NO.1 into the expression vector.

[0011] Preferably according to the present invention, the expression vector is a pET-32A expression vector.

[0012] A recombinant cell is obtained by transforming the above-mentioned recombinant expression vector into the cell.

[0013] Preferably according to the present invention, the cell is Escherichia coli BL21(DE3).

[0014] The application of the above-mentioned modified β-glucosidase in the production of laminaribiose, cellobiose and / or sophorose, the steps are as follows:

[0015] The expression gene whose nucleotide sequence is shown in SEQ ID NO.2 is introduced ...

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Abstract

The invention relates to a modified β-glucosidase, its expression gene and application. The modified β-glucosidase amino acid sequence is shown in SEQ ID NO.1; the β-glucosidase expression gene, nucleic acid sequence is shown in SEQ ID NO.2. Utilize β-glucosidase described in the present invention to produce the disaccharides such as laminaribiose, cellobiose, sophorose, compare with existing disaccharide production technology, speed of reaction is obviously quickened, and reaction temperature is gentler, has effectively reduced Production energy consumption and production cost, and the β-glucosidase of the present invention has a high expression level in the recombinant strain, the purification process is simple, and the produced laminaribiose and sophorose content is high, which is suitable for large-scale industrial production.

Description

technical field [0001] The present invention relates to a modified β-glucosidase and its expression gene and its application, especially a method for modifying β-glucosidase and its expression gene and using the enzyme to produce laminaribiose, cellobiose and sophorose , belonging to the field of biotechnology and biochemical technology. Background technique [0002] Disaccharides are sugars formed by the condensation of two monosaccharide molecules with the removal of one water molecule. Common disaccharides include sucrose, laminaribiose, cellobiose, sophorose, etc. Laminaribiose is a product of glucose caramelization, which is mostly used in the agricultural field and can be used as a preservative. Generally, it is obtained by hydrolysis or acetic acid hydrolysis of natural plant polysaccharides; cellobiose is a product of cellulose hydrolysis, and is often used as an inducer of cellulase produced by strains such as Trichoderma; sophorose is mostly used in medicine, chem...

Claims

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Application Information

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IPC IPC(8): C12N9/42C12N15/56C12N15/70C12N1/21C12P19/14C12P19/12C12R1/19
CPCC12N9/2445C12P19/12C12P19/14C12Y302/01021
Inventor 方诩牛康乐韩丽娟王明钰
Owner SHANDONG UNIV
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