Isolation and characterization of alkane hydroxylases from a metagenomic library of Pacific deep-sea sediment

Meixiang Xu, Xiang Xiao, Fengping Wang

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Two clones 9E7 and 21G8 in a metagenomic library of the east Pacific deep-sea sediment were found to contain alkane hydroxylase genes (alkB). The whole insert sequences of the two cosmid clones were determined. The insert sequences of 9E7 and 21G8 are 40 and 35 kb, respectively. Besides alkB, several alcohol/aldehyde dehydrogenase genes were also determined. A homolog of rubredoxin 2 of Pseudomonas putida was identified on 9E7 immediately downstream the alkB gene, but was lacking on 21G8. Unlike previous reports, the alkB genes on 9E7 and 21G8 have opposite transcription directions to those of linked alcohol/aldehyde dehydrogenase genes. Phylogenetic analysis put these two deep-sea AlkBs into a unique branch of integral membrane hydroxylases. The two alkB genes (9E7-alkB and 21G8-alkB) were cloned into pCom8 and introduced into two alkB expression host systems P. fluorescens KOB2Δ1 and P. putida GPo12 (pGEc47ΔB). The transformed strains can grow on the n-alkanes from C5 to C16, indicating that both 9E7-AlkB and 21G8-AlkB have a wide substrate range. The data further indicate that the deep sea would be a rich resource for exploring novel alkane-degrading strains and genes.

Original languageEnglish (US)
Pages (from-to)255-262
Number of pages8
JournalExtremophiles : life under extreme conditions
Volume12
Issue number2
DOIs
StatePublished - Mar 2008
Externally publishedYes

Fingerprint

Cytochrome P-450 CYP4A
Metagenomics
Oceans and Seas
Genes
Aldehyde Dehydrogenase
Pseudomonas putida
Alkanes
Alcohol Dehydrogenase
Rubredoxins
Clone Cells
Cosmids
Mixed Function Oxygenases
Membranes

Keywords

  • Alkane hydroxylases
  • Deep sea
  • Metagenomic library

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Microbiology

Cite this

Isolation and characterization of alkane hydroxylases from a metagenomic library of Pacific deep-sea sediment. / Xu, Meixiang; Xiao, Xiang; Wang, Fengping.

In: Extremophiles : life under extreme conditions, Vol. 12, No. 2, 03.2008, p. 255-262.

Research output: Contribution to journalArticle

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