Radioactivity resistant anomalous coccus trehalose synthase gene and trehalose making method

A technology of trehalose synthase and Deinococcus, which is applied in the direction of isomerase, genetic engineering, plant gene improvement, etc., can solve problems such as unapplied, different enzyme activities, and great differences in DNA sequence and amino acid sequence

Inactive Publication Date: 2005-01-26
南宁中诺生物工程有限责任公司
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  • Abstract
  • Description
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  • 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,...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] 1. Cloning of trehalose synthase gene (treS)

[0104] Deinococcus radiodurans was inoculated in the following liquid medium (g / L): glucose 5, yeast extract 5, peptone 10, sodium chloride (NaCl) 5, pH 7.4, Then shake on a constant temperature shaker at 30° C. for 24 hours, and centrifuge at 5000 rpm for 10 minutes to collect bacteria. Total DNA extraction of Deinococcus radiodurans was then performed as described in "Molecular Cloning: A Laboratory Manual (Second Edition)" (Sambrook, et al. 1989, Molecular Cloning: a Laboratory Manual).

[0105] Design the following primers:

[0106] 1. Upstream primer (Sense primer):

[0107] 5-GAG CCATGG CCCAGGCACACCCGGAGTGGT-3

[0108] 2. Antisense Primer

[0109] 5-TGGTCT CTGCAG TCAATTCAACCGCAGCCAGTAATAG-3

[0110] 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.

[0111] The PCR...

Embodiment 2

[0125] Using cassava starch as the starting material, after liquefaction by α-amylase, the maltose generated by the action of 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

The invention relates to gene cloning of wholly new fucose synthetase and its expression in related host. The enzyme gene is cloned from Deinococcus radioduran, which can transform the malt to fucose under normal biochemical conditions. The substrate conversion rate rises while the reaction temperature reduces. The fucose can be used in food industry, sea product cold storage and pharmaceutical activity preserving.

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): C12N1/20C12N9/90C12N15/61C12P19/12
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 南宁中诺生物工程有限责任公司
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