Protein with alpha-galactosidase activity and application of protein

A galactosidase, protein technology, applied in the application, glycosylase, enzyme and other directions, can solve the problems of affecting the digestion and absorption of nutrients, easy to produce flatulence and so on

Pending Publication Date: 2017-05-17
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This type of oligosaccharides is fermented by large intestine microorganisms, wh

Method used

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  • Protein with alpha-galactosidase activity and application of protein
  • Protein with alpha-galactosidase activity and application of protein
  • Protein with alpha-galactosidase activity and application of protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Embodiment 1, the preparation of α-galactosidase galC

[0082] The present invention provides an alpha-galactosidase gene (galC gene) derived from Aspergillus oryzae, the sequence of the galC gene is sequence 2 in the sequence listing, and its coding sequence (CDS) is sequence 3 (sequence 3 The shown DNA fragment is named as the galC coding gene), the α-galactosidase (galC) shown in the 92-817th position of the sequence 3 coding sequence 1, the sequence 3 is carried out codon optimization, and the sequence encoding galC is obtained 43-2223, the galC-encoding gene shown in the 43-2223-position of Sequence 4 is recorded as the galC-Y encoding gene.

[0083] 1. Preparation of recombinant plasmids and recombinant bacteria

[0084] The DNA fragment between the EcoR I and Xba I recognition sequences of the pPICZαA plasmid was replaced with the galC coding gene shown in Sequence 3 to obtain a recombinant plasmid, which was named pPICZαA-galC. pPICZαA-galC can express the pro...

Embodiment 2

[0123] Example 2, α-galactosidase galC can degrade oligosaccharides in soybean milk

[0124] 1. Preparation of soymilk

[0125] Weigh 100g of soybeans, soak them in tap water overnight at room temperature, put them into a soymilk machine, and crush them to produce soymilk.

[0126] 2. galC can degrade oligosaccharides in soy milk

[0127] Three treatment groups and three control groups were set up in the experiment, and each group had two parallel experiments. The three treatments were 5 U / ml, 10 U / ml, and 15 U / ml of enzyme, that is, treatment 1, treatment 2 and treatment 3. The specific operation steps are as follows:

[0128] The operation steps of treatment 1, treatment 2 and treatment 3 are as follows: after adjusting the concentration of the fermentation supernatant in step 3 of Example 1, mix it with the soymilk supernatant in equal volumes to obtain a mixed solution, and the enzyme amount of galC in the mixed solution of treatment 1 is 5U / ml, the enzyme amount of ga...

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Abstract

The invention discloses a protein with alpha-galactosidase activity and an application of the protein. The protein with alpha-galactosidase activity is any one of A1)-A5) as follows: A1) the protein with the amino acid sequence from the 92th position to the 817th position of a sequence 1 in a sequence table; A2) the protein with the amino acid sequence from the 87th position to the 817th position of the sequence 1 in the sequence table; A3) the protein with the amino acid sequence shown in the sequence 1 in the sequence table; A4) the protein with the amino acid sequence from the 87th position to the 831th position of sequence 5 in the sequence table; A5) the protein with the amino acid sequence shown in sequence 5 in the sequence table. Experiments prove that protein has alpha-galactosidase activity and the protein and a fermentation product of a recombinant microorganism containing protein-encoding nucleic acids can be used for hydrolyzing substances containing alpha-galactoside bonds and preparing alpha-galactosidase preparations.

Description

technical field [0001] The invention relates to a protein with α-galactosidase activity and its application in the field of biotechnology. Background technique [0002] α-galactosidase, also known as melibiase, can hydrolyze the terminal galactose residues of oligosaccharides such as melibiose, raffinose, and stachyose. α-galactosidase is derived from various microorganisms such as plants and bacteria, fungi, actinomycetes, and yeasts. The α-galactosidases from different sources have great differences in gene structure, molecular weight, isoelectric point, catalytic activity and so on. α-Galactosidase is a glycoprotein with a molecular weight between 30-400kDa. The protein and the glycosyl are connected by covalent bonds, and the glycosyl is mainly composed of 6-8 mannose and acetylglucosamine monosaccharides. The application fields of the enzyme mainly include the following aspects: [0003] 1. Application in food industry. Beans are the main source of protein for human...

Claims

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Application Information

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IPC IPC(8): C12N9/40C12N15/56C12P19/14C12P19/12C12P19/02
CPCC12N9/2465C12P19/02C12P19/12C12P19/14C12Y302/01022
Inventor 曹云鹤杨旭杨勇智郭贺楠郭昱含王剑王春林李德发
Owner CHINA AGRI UNIV
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