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Synthetase gene for T.fusca mycose and method for preparing mycose

A trehalose synthase, trehalose technology, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., can solve problems such as DNA sequences and amino acid sequences are very different, enzyme activities are different, and bacterial species sources are different.

Inactive Publication Date: 2005-01-12
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] Although these publications have provided some enzymes and technical solutions related to trehalose synthesis through microbial isolation and cloning, and then these enzymes are used to prepare trehalose, the strain sources of these trehalose synthases are different, and the DNA sequences and The amino acid sequence is very different, the enzyme activity is different, and most of them have not been applied at present

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0110] 1. Cloning of trehalose synthase gene (treS)

[0111] Inoculate Thermobifida fusca into the following liquid medium (g / L): glucose 4.0, yeast extract 4.0, malt extract 10.0, pH 7.2, and then shake at a constant temperature of 50°C Shake on the bed for 24 hours, then centrifuge at 5000rpm for 10 minutes to collect the bacteria. Total DNA extraction from Thermobista spp. was then performed as described in "Molecular Cloning: A Laboratory Manual (Second Edition)" (Sambrook, et al. 1989, Molecular Cloning: a Laboratory Manual).

[0112] Design the following primers:

[0113] 1. Upstream primer (Sense primer):

[0114] 5-GAG CCATGG CCCAGGCACACCCGGAGTGGT-3

[0115] 2. Antisense Primer

[0116] 5-TGGTCT CTGCAG TCAATTCAACCGCAGCCAGTAATAG-3

[0117] '

[0118] Then proceed according to the steps described in PCR Cloning Protocols: 94°C for 60 seconds, and then perform 27 ring diameters: 95°C for 45 seconds, 55°C for 60 seconds, and 72°C for 10 minutes.

[0119] The PCR ...

Embodiment 2

[0132] Using cassava starch as the starting material, after liquefaction by α-amylase, the maltose generated by β-amylase is reacted according to Example 1. The experiment shows that 100 kg of starch can obtain about 40 kg of trehalose. The conversion efficiency of moisture and impurities in starch is the same as that of using maltose directly as a substrate.

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Abstract

This invention relates to a new clone mycose synthesized zyme gene and its expression in the correlated host which zyme gene clone belong to thermobifida turning the malt sager to mycose under normal biochemical reaction, its zyme substrate conversion efficiency increases along with the decreasing of the reaction temperature, taking the malt sager or amylum as the raw material and turn it to alpha 1-mycose alpha-1 by cloned.

Description

technical field [0001] The invention relates to a trehalase gene, in particular to an enzyme obtained by using biotechnology and a method for producing trehalose. Background technique [0002] Trehalose, more precisely α,α-1,1-trehalose is composed of two molecules of glucose connected by α,α-1,1-glycosidic bonds. Trehalose is a natural disaccharide widely present in microorganisms, shrimps, Saccharomyces cerevisiae, mushrooms and various fungi, insects, plants, etc. But the content is very small. In the past, it was mainly extracted from dry yeast. Due to the low content, the extraction process is complicated and the cost is extremely high. Such expensive trehalose can only be limited to some special fields such as the application of activity maintenance of medical biological products. With the development and progress of human society, it is becoming more and more obvious to apply trehalose to maintain the umami taste and improve the texture of various foods. Therefore...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/90C12N15/61C12P19/12
Inventor 韦宇拓黄日波蒙健宗卢福燊庞中文朱绮霞陈发忠罗兆飞卢运琨王青艳黄鲲
Owner GUANGXI UNIV
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