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Screening of Organic Tolerant Organisms and Homologous Expression of Lipase Gene in Burkholderia sp.SIB005

Author: JiangHuan
Tutor: HuangJianZhong
School: Fujian Normal University
Course: Microbiology
Keywords: An organic solvent Homologous expression Lipase
CLC: Q93
Type: Master's thesis
Year: 2010
Downloads: 41
Quote: 0
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Abstract


Solvent Tolerant Bacteria are a class of novel extremophiles , until the 1980s was only system in-depth study . Through a variety of tolerance mechanisms , effectively resist or reduce the toxic effects of organic solvents their cells . Therefore, in the whole-cell catalysis, environmental pollution control , resistance to organic solvents extremophiles have broad prospects for industrial applications . In this study, through targeted screening a the lipase production capacity and benzene and alkanes with organic solvent tolerance Burkholderia species Burkholderia sp.SIB005 view of Burkholderia strains fat the complex enzyme secretion mechanism and the need for homologous chaperone protein involved in the experimental choose by homologous to express the way to further improve the vitality of the of Burkholderia sp.SIB005 lipase . In order to achieve the lipase gene of the strain on the efficient expression of the clone the Burkholderia sp.SIB005 the lipase gene and the partner gene and upstream promoter region ( lipAB ) and the promoter region of The Burkholderia sp.SIB005 lipase gene and upstream ( LIPA ) with low copy expression the plasmid pBBR22b connection was transformed into E. coli DH5a , plasmids were extracted through electricity into the two recombinant the expression plasmid go to SIB005 strain was successfully constructed containing pBBR22b-lipAB, pBBR22b-lipA engineered strains of the two expression plasmids . The two engineering strain and strain SIB005 fat enzyme activity was measured under the same pieces By comparison, the first two lipase activity were increased by 88 % and 25% of highest SIB005 of initial enzyme activity on the basis of experiments show that the lipase genes highly expressed the lipase chaperone protein inseparable . By increasing the number of copies of the lipase gene and the chaperone gene and upstream promoter , the successful implementation of the efficient expression of homologous the strain SIB005 lipase .

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CLC: > Biological Sciences > Microbiology
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