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Studies on Alkaliphilic Mechnism of Alkaliphilic β-Mannanase from Alkaliphilic Bacillus SP.16-5

Author: ZhangYunHua
Tutor: DanAnShan
School: Northeast Agricultural University
Course: Animal Nutrition and Feed Science
Keywords: Mechanism of basophils β - mannanase Structure and Function Error-prone PCR
CLC: Q936
Type: Master's thesis
Year: 2004
Downloads: 135
Quote: 3
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Abstract


Mechanism to maintain the vitality and stability of proteins under alkaline conditions in protein engineering research hot spots , but due to the complexity of the mechanism involved in the research , little alkalophilic Bacillus N16-5 basophilic the β - mannanase of research material , the preliminary study on the the alkalophilic mechanism protein under alkaline conditions to maintain the vitality and stability . The study basophilic β - mannanase gene isolated from alkalophilic Bacillus N16-5 genome subcloned pUC18 for cloning and expression vector was constructed mutant library . DCTP and dTTP the - rich dNTF mixture containing high concentrations of Mg 2 < / sup> PCR buffer solution was subjected to error-prone PCR to construct a mutant library by high throughput screening method , filtering, to obtain an optimum pH mutant changed , a structure and spatial changes in the structure of the measurement and prediction , a structure determination showed that changes in the two mutation points in total , in order to analyze the variation in the characteristics of the enzyme caused by the point of change of a single bit result , or both together , to build a single mutant chimeras . The results showed that the glycine (A) to valine (V) changes in the optimum pH of the mutant enzyme molecules changed . Prediction of the spatial structure of that may be due to the two spiral decrease of the molecular surface , resulting in the variation in characteristics . Alkalophilic enzyme on a structure from a structural comparison of the alkali - β- mannanase and other β - mannanase of the addicted , and no uniform characteristics to explain their basophils .

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CLC: > Biological Sciences > Microbiology > Microbial biochemistry
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