Cloning and expressing DBT (dibenzothiophene) monooxygenase gene (dszC) from Rhodococcus sp. DS-3 in Escherichia coli |
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Authors: | Ma Ting Li Shanshan Li Guoqiang Wang Renjing Liang Fenglai and Liu Rulin |
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Institution: | (1) College of Life Sciences, Nankai University, Tianjin, 300071, China |
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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 (DBTO2). 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 Ni2+ 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.
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Translated from Acta Scientiarum Naturalium Universitatis Nankaiensis, 2005, 38(6): 1–6 译自: 南开大学学报 (自然科学版), 2005, 38(6): 1–6] |
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Keywords: | biodesulfurization Rhodococcus sp dibenzothiophene monooxidase expression |
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