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Cloning of Baeyer-Villiger monooxygenases from Comamonas, Xanthobacter and Rhodococcus using polymerase chain reaction with highly degenerate primers by Jan B. Van Beilen; Frédéric Mourlane; Markus A. Seeger; Jasminka Kovac; Zhi Li; Theo H. M. Smits; Urban Fritsche; Bernard Witholt is a Environmental Science article available to read on EtoBox.

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## Summary To clone novel type 1 Baeyer‐Villiger monooxygenase (BVMO) genes, we isolated or collected 25 bacterial strains able to grow on alicyclic compounds. Twelve of the bacterial strains yielded polymerase chain reaction (PCR) fragments with highly degenerate primers based on the sequences of known and putative BVMOs. All these fragments were found to encode peptides homologous to published BVMO sequences. The complete BVMO genes and flanking DNA were cloned from a __Comamonas__, a __Xanthobacter__ and a __Rhodococcus__ strain using the PCR fragments as probes. BVMO genes cloned from the first two strains could be expressed to high levels in __Escherichia coli__ using standard expression vectors, and the recombinants converted cyclopentanone and cyclohexanone to the corresponding lactones. The __Rhodococcus__ BVMO, a putative steroid monooxygenase, could be expressed after modification of the N‐terminal sequence. However, recombinants expressing this protein did not show activity towards progesterone. An esterase homologue located directly upstream of the __Xanthobacter__ BVMO gene and a dehydrogenase homologue encoded directly downstream of the __Comamonas__ sp. NCIMB 9872 BV

Who reads Cloning of Baeyer-Villiger monooxygenases from Comamonas, Xanthobacter and Rhodococcus using polymerase chain reaction with highly degenerate primers?

Author

Jan B. Van Beilen; Frédéric Mourlane; Markus A. Seeger; Jasminka Kovac; Zhi Li; Theo H. M. Smits; Urban Fritsche; Bernard Witholt

Publisher

John Wiley and Sons; Wiley (Blackwell Publishing); Blackwell Publishing Inc.; Wiley (ISSN 1462-2912)

Published

2003

Language

EN

Field

Environmental Science (Physical Sciences)