Improvement of desulfurization activity in Rhodococcus erythropolis KA2-5-1 by genetic engineering

• Published in: Bioscience, biotechnology, and biochemistry Vol. 65; no. 2; pp. 239 - 246
• Main Authors: Hirasawa, K. (Petroleum Energy Center, Shimizu, Shizuoka (Japan)), Ishii, Y, Kobayashi, M, Koizumi, K, Maruhashi, K
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience Biotechnology and Agrochemistry 01.02.2001
• Subjects: Amino Acid Sequence; Bacterial Proteins - genetics; Biological and medical sciences; Biotechnology; dibenzothiophene; DNA Helicases; DNA-Binding Proteins; dszA gene; dszB gene; dszC gene; Energy; ENZYMIC ACTIVITY; Escherichia coli; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Genes, Bacterial; GENETIC ENGINEERING; Genetic technics; Genetic Vectors; Industrial applications and implications. Economical aspects; Methods. Procedures. Technologies; Modification of gene expression level; Molecular Sequence Data; Multigene Family; plasmid pRC4; Plasmids - genetics; POLLUTION; Proteins - genetics; RHODOCOCCUS (BACTERIA); Rhodococcus - genetics; Rhodococcus - metabolism; Rhodococcus erythropolis; Sequence Homology, Amino Acid; sulfur; Sulfur - metabolism; Thiophenes - metabolism; Trans-Activators; Use of microorganisms or biopolymers in fossil fuel recovery; Light gas oil; Recombinant microorganism; Plasmid; Gene cluster; Desulfurization; Thiophene derivatives; Genetic transformation; Sulfur Organic compounds; Bacteria; Rhodococcus erythropolis; Actinomycetes; Fuel desulfurization
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.65.239

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.