Functional Analysis of Alkane Hydroxylases from Gram-Negative and Gram-Positive Bacteria
TLDR
Three recombinant host strains are developed for the functional analysis of the novel alkane hydroxylase genes and sequence comparisons show that the level of protein sequence identity between the homologs is as low as 35%, and that the Pseudomonas alkane Hydroxylases are as distantly related to each other as to the remaining alkanes.Abstract:
We have cloned homologs of the Pseudomonas putida GPo1 alkane hydroxylase from Pseudomonas aeruginosa PAO1, Pseudomonas fluorescens CHA0, Alcanivorax borkumensis AP1, Mycobacterium tuberculosis H37Rv, and Prauserella rugosa NRRL B-2295. Sequence comparisons show that the level of protein sequence identity between the homologs is as low as 35%, and that the Pseudomonas alkane hydroxylases are as distantly related to each other as to the remaining alkane hydroxylases. Based on the observation that rubredoxin, an electron transfer component of the GPo1 alkane hydroxylase system, can be replaced by rubredoxins from other alkane hydroxylase systems, we have developed three recombinant host strains for the functional analysis of the novel alkane hydroxylase genes. Two hosts, Escherichia coli GEc137 and P. putida GPo12, were equipped with pGEc47 Delta B, which encodes all proteins necessary for growth on medium-chain-length alkanes (C(6) to C(12)), except a functional alkane hydroxylase. The third host was an alkB knockout derivative of P. fluorescens CHA0, which is no longer able to grow on C(12) to C(16) alkanes. All alkane hydroxylase homologs, except the Acinetobacter sp. ADP1 AlkM, allowed at least one of the three hosts to grow on n-alkanes.read more
Citations
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Journal ArticleDOI
Obligate oil-degrading marine bacteria.
TL;DR: Genome sequencing and functional genomic analysis of Alcanivorax borkumensis, the paradigm of OHCB, has provided significant insights into the genomic basis of the efficiency and versatility of its hydrocarbon utilization, the metabolic routes underlying its special hydrocarbon diet, and its ecological success.
Journal ArticleDOI
Degradation of alkanes by bacteria
TL;DR: This review summarizes current knowledge on how microorganisms degrade alkanes, focusing on the biochemical pathways used and on how the expression of pathway genes is regulated and integrated within cell physiology.
Journal ArticleDOI
Alkane hydroxylases involved in microbial alkane degradation.
TL;DR: This review focuses on the role and distribution in the environment of alkane hydroxylases and their (potential) applications in bioremediation and biocatalysis.
Journal ArticleDOI
Genome sequence of the ubiquitous hydrocarbon-degrading marine bacterium Alcanivorax borkumensis
Susanne Schneiker,Vitor A. P. Martins dos Santos,Daniela Bartels,Thomas Bekel,Martina Brecht,Jens Buhrmester,Tatyana N. Chernikova,Renata Denaro,Manuel Ferrer,Christoph Gertler,Alexander Goesmann,Olga V. Golyshina,Filip Kaminski,Amit N. Khachane,Siegmund Lang,Burkhard Linke,Alice C. McHardy,Alice C. McHardy,Folker Meyer,Taras Nechitaylo,Alfred Pühler,Daniela Regenhardt,Oliver Rupp,Julia S. Sabirova,Werner Selbitschka,Michail M. Yakimov,Kenneth N. Timmis,Frank-Jörg Vorhölter,Stefan Weidner,Olaf Kaiser,Peter N. Golyshin +30 more
TL;DR: The genome sequence provides the basis for the future design of strategies to mitigate the ecological damage caused by oil spills and provides A. borkumensis SK2 with a competitive edge in oil-polluted environments.
Journal ArticleDOI
Bacterial metabolism of long-chain n-alkanes.
Alexander Wentzel,Alexander Wentzel,Trond E. Ellingsen,Hans-Kristian Kotlar,Sergey B. Zotchev,Mimmi Throne-Holst +5 more
TL;DR: Recent advances in the understanding of bacterial metabolism of long-chain n-alkanes are summarized and bacterial strategies for accessing these highly hydrophobic substrates are presented, along with systems for their enzymatic degradation and conversion into products of potential industrial value.
References
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Arapid alkaline extraction procedure forscreening recombinant plasmid DNA
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