Two sets of biphenyl and PCB degradation genes on a linear plasmid in Rhodococcus erythropolis TA421

Polychlorinated Biphenyl/Biphenyl Degrading Gene Clusters inRhodococcussp. K37, HA99, and TA431 Are Different from Well-KnownbphGene Clusters of Rhodococci

“…TA421 has seven bphC genes (bphC1 to bphC7). 10,11) In addition, TA421 carries an approximately 500-kb linear plasmid that contains three of the seven bphC genes (bphC2, bphC3, and bphC4).…”

Functional Expression of Three Rieske Non-Heme Iron Oxygenases Derived from Actinomycetes inRhodococcusSpecies for Investigation of Their Degradation Capabilities of Dibenzofuran and Chlorinated Dioxins

“…For promoter replacement (i.e., to switch from P dfdA to P dfdB ), the dfdA1-A4 genes with EcoRV sites introduced just downstream of the start codon of dfdA1 were PCR-amplified and ligated downstream of P dfdB (pRK401-P B +dfdA). A plasmid, pTB1, that encoded bphA1A2A3A4 of R. erythropolis TA421 was used as the PCR template, 28) and the amplified DNA fragment extending from the start codon of bphA1 TA421 to the stop codon of bphA4 TA421 was ligated downstream of P dfdB (pRK401-P B +bphA TA421 ). The two functionally unidentified genes in the bph gene cluster of R. rhodochrous K37 23) separating the terminal dioxygenase genes bphA1A2 from the electron transfer protein genes bphA3A4 were removed.…”

“…42) It has been reported that S. wittichii RW1 has multiple copies of the terminal dioxygenase subunit gene for AhDO, 43) and there is a possibility that these uninvestigated AhDOs affect dioxin degradation in RW1 cells. Also, it has been shown that R. opacus SAO101 can utilize diverse aromatic compounds, 44) and therefore might have multiple AhDO genes with different specificities in addition to narAaAb genes, 45) as reported for some diverse aromatics-degrading actinomycetes, such as Rhodococcus jostii RHA1, 46) R. erythropolis TA421, 28) and Mycobacterium vanbaalenii PYR-1.…”

“…YK3 20) and the biphenyl (BP)-utilizing strains Rhodococcus erythropolis TA421 28) and R. rhodochrous K37. 23) In this study, three actinomycetales-derived AhDO genes for which it is difficult to produce functionally active proteins in E. coli were functionally expressed in a Rhodococcus strain using a constitutive expression promoter for the genus Rhodococcus.…”

“…We have reported that both of these genes were involved in BP utilization. 23,28) The amino acid sequences of their respective BphA subunits had only from 35% (BphA4) to 44% (BphA3) identity. Our preliminary attempts to express the two BphAs in E. coli failed (data not shown for BphA TA421 , see ref.…”