Thermostabilization of desulfurization enzymes from Rhodococcos sp. IGTS8. Final technical report

II, John J. Kilbane

Gas Technology Institute, Des Plaines, IL (United States)

关键词: Rhodococcus;    02 Petroleum;    Clone Cells;    Vectors Rhodococcus;    Oxidoreductases;   

DOI  :  10.2172/809375
RP-ID  :  NONE
RP-ID  :  FG02-97ER62464
RP-ID  :  809375

美国|英语

来源: UNT Digital Library

【 摘 要 】

The objective of this project was to develop thermophilic cultures capable of expressing the desulfurization (dsz) operon of Rhodococcus sp. IGTS8. The approaches taken in this project included the development of plasmid and integrative expression vectors that function well in Thermus thermophilus, the cloning of Rhodococcus dsz genes in Thermus expression vectors, and the isolation of bacterial cultures that express the dsz operon at thermophilic temperatures. This project has resulted in the development of plasmid and integrative expression vectors for use in T. thermophilus. The dsz genes have been expressed at moderately thermophilic temperatures (52 C) in Mycobacterium phlei and at temperatures as high as 72 C in T. thermophilus. The tools and methods developed in this project will be generally useful for the expression of heterologous genes in Thermus. Key developments in the project have been the isolation of a Mycobacterium phlei culture capable of expressing the desulfurization operon at 52 C, development of plasmid and integrative expression vectors for Thermus thermophilus, and the development of a host-vector system based on the malate dehydrogenase gene that allows plasmids to be stably maintained in T. thermophilus and provides a convenient reporter gene for the accurate quantification of gene expression. Publications have been prepared regarding each of these topics; these preprints are included.