Improvement of desulfurization activity in Rhodococcus erythropolis KA2-5-1 by genetic engineering.
Rhodococcus erythropolis KA2-5-1 can desulfurize dibenzothiophene (DBT) into 2-hydroxybiphenyl. A cryptic plasmid, pRC4, which was derived from R. rhodochrous IFO3338, was combined with an Escherichia coli vector to construct an E. coli-Rhodococcus shuttle vector. The complete nucleotide sequence of 2582-bp pRC4 was analyzed. Based on the characteristics of its putative replication genes, pRC4 was assigned to the family of pAL5000-related replicons. The desulfurization gene cluster, dszABC, and the related reductase gene, dszD, cloned from KA2-5-1, were reintroduced into KA2-5-1 and efficiently expressed. The DBT desulfurization ability of the transformant carrying two dszABC clusters and one dszD on the vector was about 4-fold higher than that of the parent strain, and the transformant also showed improved desulfurization activity for light gas oil (LGO). Sulfur components in LGO before and after the reaction were analyzed with gas chromatography-atomic emission detection.
Hirasawa
K
K
Bio-Refining Process Laboratory, Advanced Technology and Research Institute, Petroleum Energy Center, Shimizu-shi, Shizuoka, Japan.
Ishii
Y
Y
Kobayashi
M
M
Koizumi
K
K
Maruhashi
K
K
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D004265:DNA Helicases; D004268:DNA-Binding Proteins; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D005818:Genetic Engineering; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D005810:Multigene Family; D010957:Plasmids / Q000235:genetics; D011506:Proteins / Q000235:genetics; D012240:Rhodococcus / Q000235:genetics* / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013455:Sulfur / Q000378:metabolism*; D013876:Thiophenes / Q000378:metabolism; D015534:Trans-Activators
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D011506:Proteins; C048817:RepB protein, Bacteria; D013876:Thiophenes; D015534:Trans-Activators; C045264:replication initiator protein; D013455:Sulfur; D004265:DNA Helicases; C016366:dibenzothiophene