Cloning, expression and characterization of a putative 7alpha-hydroxysteroid dehydrogenase in Comamonas testosteroni. 2014

Wei Ji, and Yuanan Chen, and Hao Zhang, and Xiao Zhang, and Ziyi Li, and Yuanhua Yu
Jilin University, College of Animal Science and Veterinary Medicine, Changchun, PR China; Changchun University of Science and Technology, School of Life Science and Technology, Changchun, PR China.

The short-chain dehydrogenase/reductase (SDR) superfamily is a large and diverse group of genes with members found in all forms of life. Comamonas testosteroni (C. testosterone) ATCC11996 is a Gram-negative bacterium which can use steroids as carbon and energy source. In the present investigation, we found a novel SDR gene 7alpha-hydroxysteroid dehydrogenase (7α-HSD) which is located 11.9 kb upstream from hsdA with the same transcription orientation in the C. testosteroni genome. The open reading frame of this putative 7alpha-hydroxysteroid dehydrogenase gene consists of 771 bp and translates into a protein of 256 amino acids. Two consensus sequences of the SDR superfamily were found, an N-terminal Gly-X-X-X-Gly-X-Gly cofactor-binding motif and a Tyr-X-X-X-Lys segment (residues 161-165 in the 7α-HSD sequence) essential for catalytic activity of SDR proteins. To produce purified 7α-HSD protein, the 7α-HSD gene was cloned into plasmid p(ET-15b) and the over expressed protein was purified by His-tag sequence on metal chelate chromatography. To prove that 7α-HSD is involved in the metabolic pathway of steroid compounds, we constructed a 7α-HSD knock-out mutant of C. testosteroni. Compared to the wild type C. testosteroni, degradation of testosterone, estradiol and cholesterol were decreased in the 7α-HSD knock-out mutant. Furthermore, growth in the medium with testosterone, estradiol and cholesterol was impaired in 7α-HSD knock-out mutant. The results showed that 7α-HSD is involved in steroid degradation.

UI MeSH Term Description Entries
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D003001 Cloning, Molecular The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells. Molecular Cloning
D006913 Hydroxysteroid Dehydrogenases Enzymes of the oxidoreductase class that catalyze the dehydrogenation of hydroxysteroids. (From Enzyme Nomenclature, 1992) EC 1.1.-. Hydroxysteroid Dehydrogenase,Dehydrogenase, Hydroxysteroid,Dehydrogenases, Hydroxysteroid
D000595 Amino Acid Sequence The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION. Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D001426 Bacterial Proteins Proteins found in any species of bacterium. Bacterial Gene Products,Bacterial Gene Proteins,Gene Products, Bacterial,Bacterial Gene Product,Bacterial Gene Protein,Bacterial Protein,Gene Product, Bacterial,Gene Protein, Bacterial,Gene Proteins, Bacterial,Protein, Bacterial,Proteins, Bacterial
D001483 Base Sequence The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence. DNA Sequence,Nucleotide Sequence,RNA Sequence,DNA Sequences,Base Sequences,Nucleotide Sequences,RNA Sequences,Sequence, Base,Sequence, DNA,Sequence, Nucleotide,Sequence, RNA,Sequences, Base,Sequences, DNA,Sequences, Nucleotide,Sequences, RNA
D015964 Gene Expression Regulation, Bacterial Any of the processes by which cytoplasmic or intercellular factors influence the differential control of gene action in bacteria. Bacterial Gene Expression Regulation,Regulation of Gene Expression, Bacterial,Regulation, Gene Expression, Bacterial
D016415 Sequence Alignment The arrangement of two or more amino acid or base sequences from an organism or organisms in such a way as to align areas of the sequences sharing common properties. The degree of relatedness or homology between the sequences is predicted computationally or statistically based on weights assigned to the elements aligned between the sequences. This in turn can serve as a potential indicator of the genetic relatedness between the organisms. Sequence Homology Determination,Determination, Sequence Homology,Alignment, Sequence,Alignments, Sequence,Determinations, Sequence Homology,Sequence Alignments,Sequence Homology Determinations
D020604 Comamonas testosteroni A species of gram-negative, aerobic rods formerly called Pseudomonas testosteroni. It is differentiated from other Comamonas species by its ability to assimilate testosterone and to utilize phenylacetate or maleate as carbon sources. Pseudomonas testosteroni

Related Publications

Wei Ji, and Yuanan Chen, and Hao Zhang, and Xiao Zhang, and Ziyi Li, and Yuanhua Yu
June 2015, Chemico-biological interactions,
Wei Ji, and Yuanan Chen, and Hao Zhang, and Xiao Zhang, and Ziyi Li, and Yuanhua Yu
June 2000, Biochemical and biophysical research communications,
Wei Ji, and Yuanan Chen, and Hao Zhang, and Xiao Zhang, and Ziyi Li, and Yuanhua Yu
November 2021, Molecular biology reports,
Wei Ji, and Yuanan Chen, and Hao Zhang, and Xiao Zhang, and Ziyi Li, and Yuanhua Yu
March 2012, The Journal of steroid biochemistry and molecular biology,
Wei Ji, and Yuanan Chen, and Hao Zhang, and Xiao Zhang, and Ziyi Li, and Yuanhua Yu
April 2005, Beijing da xue xue bao. Yi xue ban = Journal of Peking University. Health sciences,
Wei Ji, and Yuanan Chen, and Hao Zhang, and Xiao Zhang, and Ziyi Li, and Yuanhua Yu
January 1999, Advances in experimental medicine and biology,
Wei Ji, and Yuanan Chen, and Hao Zhang, and Xiao Zhang, and Ziyi Li, and Yuanhua Yu
February 2003, Chemico-biological interactions,
Wei Ji, and Yuanan Chen, and Hao Zhang, and Xiao Zhang, and Ziyi Li, and Yuanhua Yu
January 2001, Chemico-biological interactions,
Wei Ji, and Yuanan Chen, and Hao Zhang, and Xiao Zhang, and Ziyi Li, and Yuanhua Yu
November 1996, European journal of biochemistry,
Copied contents to your clipboard!