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Purificiation of Cryptococcus Laurentii Lipase and Rapid Measurement of Lipase Thermostability

Author: FengWeiZong
Tutor: LinLin
School: Fujian Normal University
Course: Biochemistry and Molecular Biology
Keywords: Cryptococcus laurentii Lipase Thermal stability ANS Purification Fluorescent probe N - terminal sequencing Enzymatic Properties
CLC: Q78
Type: Master's thesis
Year: 2010
Downloads: 36
Quote: 0
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Abstract


Lipase as an important industrial enzymes, paper, pharmaceutical , food , household chemicals, energy development and other fields have a wide range of research and application . By ion-exchange column Sepharose Fast Flow to purified of Cryptococcus laurentii Yeast B40 ( Cryptococcus laurentii B40 ) , lipase , a purity of more than 95% of the lipase solution , SDS - PAGE electrophoresis shows the relative molecular mass of about 44.3KDa sequence analysis showed that the N-terminal 12 amino acid sequence of the lipase : DFGPITIYTPPA . Enzymatic properties analysis showed that the lipase is a low-temperature alkaline lipase , more suited to a temperature of 25 ° C , the optimum pH of 10.2 , the Tm to 40.90 ° C , able to catalyze the hydrolysis of short carbon chain ( 12 carbon ) triglycerides, mixed carbon-based triglycerides also have a catalytic role . Analysis of the influence of the type of metal ion and a surfactant of the enzyme , the heavy metal ions can promote lipid was dissolved in the reagent is able to improve the catalytic effectiveness of the strong inhibition of the enzyme activity play . The thermal stability of the lipase is lipase one of the important properties , good thermal stability means save for convenience, ease of transport, be able to withstand the high temperature environment in the industrial production applications . Looking for a simple and rapid determination of thermal stability of the lipase method for screening thermal stability of lipase plays an important role , which can greatly save the cost of experiments to improve the efficiency of the experiment , to shorten the cycle of research into the production practice . This paper reports a simple, high-throughput thermal stability assay of lipase - ANS lipase rapid determination of the stability of the establishment of the method , the method is the measurement time is shortened by a few days of the traditional NaOH titration hours and is very suitable for high-throughput determination of thermal stability of lipase .

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