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Fucose synthetase gene of corynebacterium glutamicium, and method for preparing fucose

A technology of trehalose synthase and Corynebacterium glutamicum, which is applied in the direction of bacteria, isomerase, DNA/RNA fragments, etc., can solve the problems of not being applied, different enzyme activities, and different sources of strains, etc.

Inactive Publication Date: 2005-01-12
南宁中诺生物工程有限责任公司
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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

[0112] 1. Cloning of trehalose synthase gene (treS)

[0113] Inoculate Corynebacterium glutamicum into the following liquid medium (g / L): 10 grams of beef extract, 10 grams of peptone, 5 grams of sodium chloride, pH 7.2-7.4, and then keep the temperature at 28-30°C Shake on a shaker for 18-36 hours, centrifuge at 5000rpm for 10 minutes to collect the bacteria, then shake on a constant temperature shaker at 30°C for 24 hours, and centrifuge at 5000rpm for 10 minutes to collect the bacteria. Total DNA extraction of Corynebacterium acidicum was then performed as described in "Molecular Cloning: A Laboratory Manual (Second Edition)" (Sambrook, et al. 1989, Molecular Cloning: a Laboratory Manual).

[0114] Design the following primers:

[0115] 1. Forward primer (Sense primer):

[0116] 5'-AGA CCATGG AAACCTCCTGGCATTTCT-3'

[0117] 2. Antisense Primer:

[0118] 5'-TCT CTGCAG TCATTCCATATCGTCCTTTTCATCG-3' is then carried out according to the steps described in PCR Cloning Proto...

Embodiment 2

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

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Abstract

This invention relates to a new mycose synthesized zyme gene clone and its expression in correlate host which zyme gene clone is from corynebacterium glutamicum CICC 10226 which can turn malt sager to mycose under normal biochemical reaction, its substrate conversion efficiency increases along with the reduction of the reaction temperature. It can take malt sager or amylum as the raw material to be turned to 1-mycose with the cloned zyme alpha alpha-1.

Description

technical field [0001] The invention relates to a method for preparing trehalose, in particular to a method for producing trehalose by using enzymes obtained by biotechnology. 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 foo...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N9/90C12N15/52C12P19/12C12P21/02
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 南宁中诺生物工程有限责任公司
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