Indene bioconversion by a toluene inducible dioxygenase of Rhodococcus sp I24

Priefert, Horst, O'Brien, Xian M., Lessard, Philip A., Dexter, Annette F., Choi, Ellen E., Tomic, Sladjana, Nagpal, Geeta, Cho, Jennie J., Agosto, Melina, Yang, Lucy, Treadway, Sheri L., Tamashiro, Lance, Wallace, Matthew and Sinskey, Anthony J. (2004) Indene bioconversion by a toluene inducible dioxygenase of Rhodococcus sp I24. Applied Microbiology And Biotechnology, 65 2: 168-176. doi:10.1007/s00253-004-1589-3

Author Priefert, Horst
O'Brien, Xian M.
Lessard, Philip A.
Dexter, Annette F.
Choi, Ellen E.
Tomic, Sladjana
Nagpal, Geeta
Cho, Jennie J.
Agosto, Melina
Yang, Lucy
Treadway, Sheri L.
Tamashiro, Lance
Wallace, Matthew
Sinskey, Anthony J.
Title Indene bioconversion by a toluene inducible dioxygenase of Rhodococcus sp I24
Journal name Applied Microbiology And Biotechnology   Check publisher's open access policy
ISSN 0175-7598
Publication date 2004-08
Sub-type Article (original research)
DOI 10.1007/s00253-004-1589-3
Volume 65
Issue 2
Start page 168
End page 176
Total pages 9
Place of publication Berlin, Germany
Publisher Springer
Language eng
Subject 1007 Nanotechnology
Abstract Rhodococcus sp. I24 can oxygenate indene via at least three independent enzyme activities: (i) a naphthalene inducible monooxygenase (ii) a naphthalene inducible dioxygenase, and (iii) a toluene inducible dioxygenase (TID). Pulsed field gel analysis revealed that the I24 strain harbors two megaplasmids of similar to340 and similar to50 kb. Rhodococcus sp. KY1, a derivative of the I24 strain, lacks the similar to340 kb element as well as the TID activity. Southern blotting and sequence analysis of an indigogenic, I24-derived cosmid suggested that an operon encoding a TID resides on the similar to340 kb element. Expression of the tid operon was induced by toluene but not by naphthalene. In contrast, naphthalene did induce expression of the nid operon, encoding the naphthalene dioxygenase in I24. Cell free protein extracts of Escherichia coli cells expressing tidABCD were used in HPLC-based enzyme assays to characterize the indene bioconversion of TID in vitro. In addition to 1-indenol, indene was transformed to cis-indandiol with an enantiomeric excess of 45.2% of cis-(1S,2R)-indandiol over cis-(1R,2S)-indandiol, as revealed by chiral HPLC analysis. The K-m of TID for indene was 380 muM. The enzyme also dioxygenated naphthalene to cis-dihydronaphthalenediol with an activity of 78% compared to the formation of cis-indandiol from indene. The K-m of TID for naphthalene was 28 muM. TID converted only trace amounts of toluene to 1,2-dihydro-3-methylcatechol after prolonged incubation time. The results indicate the role of the tid operon in the bioconversion of indene to 1-indenol and cis-(1S,2R)-indandiol by Rhodococcus sp. I24.
Keyword Biotechnology & Applied Microbiology
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Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
Australian Institute for Bioengineering and Nanotechnology Publications
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Created: Fri, 25 Jan 2008, 16:18:35 EST