NDLI: Cloning and expressing DBT (dibenzothiophene) monooxygenase gene (dszC) from Rhodococcus sp. DS-3 in Escherichia coli

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Cloning and expressing DBT (dibenzothiophene) monooxygenase gene (dszC) from Rhodococcus sp. DS-3 in Escherichia coli

Content Provider	Springer Nature Link
Author	Ma, Ting Li, Shanshan Li, Guoqiang Wang, Renjing Liang, Fenglai Liu, Rulin
Copyright Year	2006
Abstract	Dibenzothiophene (DBT) monooxygenase (DszC) catalysis, the first and also the key step in the microbial DBT desulfurization, is the conversion of DBT to DBT sulfone (DBTO$_{2}$). In this study, dszC of a DBT-desulfurizing bacterium Rhodococcus sp. DS-3 was cloned by PCR. The sequence cloned was 99% homologous to Rhodococcus erythropolis IGTS8 that was reported in the Genebank. The gene dszC could be overexpressed effectively after being inserted into plasmid pET28a and transformed into E. coli BL21 strain. The expression amount of DszC was about 20% of total supernatant at low temperature. The soluble DszC in the supernatant was purified by Ni$^{2+}$ chelating His-Tag resin column and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to electronics purity. Only one band was detected by Western-blotting, which is for the antibody released in mouse against purified DszC in the expression product of BL21 (DE3, paC5) and Rhodococcus sp. DS-3. The activity of purified DszC was 0.36 U. DszC can utilize the organic compound such as DBT and methyl-DBT, but not DBT derivates such as DBF, which has no sulfur or inorganic sulfur.
Starting Page	375
Ending Page	380
Page Count	6
File Format	PDF
ISSN	16733509
Journal	Frontiers in Biology
Volume Number	1
Issue Number	4
e-ISSN	16733622
Language	English
Publisher	Nauka/Interperiodica
Publisher Date	2006-01-01
Publisher Institution	Chinese Universities
Publisher Place	Moscow
Access Restriction	Subscribed
Subject Keyword	biodesulfurization Rhodococcus sp dibenzothiophene monooxidase expression Life Sciences
Content Type	Text
Resource Type	Article
Subject	Ecology Genetics Ecology, Evolution, Behavior and Systematics Biotechnology

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