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

Meixiang Xu; Xiang Xiao; Fengping Wang

doi:10.1007/s00792-007-0122-x

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 journal › Article › peer-review

37 Scopus citations

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 language	English (US)
Pages (from-to)	255-262
Number of pages	8
Journal	Extremophiles
Volume	12
Issue number	2
DOIs	https://doi.org/10.1007/s00792-007-0122-x
State	Published - Mar 2008
Externally published	Yes

Keywords

Alkane hydroxylases
Deep sea
Metagenomic library

ASJC Scopus subject areas

Microbiology
Molecular Medicine

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10.1007/s00792-007-0122-x

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@article{2322db8c610b4c87b81a87437ca54a63,

title = "Isolation and characterization of alkane hydroxylases from a metagenomic library of Pacific deep-sea sediment",

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.",

keywords = "Alkane hydroxylases, Deep sea, Metagenomic library",

author = "Meixiang Xu and Xiang Xiao and Fengping Wang",

note = "Funding Information: Acknowledgments We would like to thank Dr. Jan B van Beilen for generously providing us the alkB gene expressed system including strains and vector of P. putida GPo1, P. putida GPo12 (pGEc47∆B), P. fluorescens KOB2∆1, E. coli RK600 and pCom8. We are indebted to Dr. Zongze Shao for his kind recommendations and helpful discussions. Our thanks are due to the crew on the R/V Dayang Yihao for help in collecting the samples. This work was supported by the NSF China Award (40476054), COMRA fund (DY7000M-02).",

year = "2008",

month = mar,

doi = "10.1007/s00792-007-0122-x",

language = "English (US)",

volume = "12",

pages = "255--262",

journal = "Extremophiles",

issn = "1431-0651",

publisher = "Springer Japan",

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T1 - Isolation and characterization of alkane hydroxylases from a metagenomic library of Pacific deep-sea sediment

AU - Xu, Meixiang

AU - Xiao, Xiang

AU - Wang, Fengping

N1 - Funding Information: Acknowledgments We would like to thank Dr. Jan B van Beilen for generously providing us the alkB gene expressed system including strains and vector of P. putida GPo1, P. putida GPo12 (pGEc47∆B), P. fluorescens KOB2∆1, E. coli RK600 and pCom8. We are indebted to Dr. Zongze Shao for his kind recommendations and helpful discussions. Our thanks are due to the crew on the R/V Dayang Yihao for help in collecting the samples. This work was supported by the NSF China Award (40476054), COMRA fund (DY7000M-02).

PY - 2008/3

Y1 - 2008/3

N2 - 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.

AB - 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.

KW - Alkane hydroxylases

KW - Deep sea

KW - Metagenomic library

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U2 - 10.1007/s00792-007-0122-x

DO - 10.1007/s00792-007-0122-x

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SN - 1431-0651

VL - 12

SP - 255

EP - 262

JO - Extremophiles

JF - Extremophiles

IS - 2

ER -

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

Abstract

Keywords

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