alexa Cloning of poly(3-hydroxybutyrate) depolymerase from a marine bacterium, Alcaligenes faecalis AE122, and characterization of its gene product
Environmental Sciences

Environmental Sciences

Journal of Pollution Effects & Control

Author(s): Keiko Kita, Nobuo Kato, Shunichiro Mashiba, Masatoshi Nagita, Kaori Ishimaru

Abstract Share this page

A DNA fragment that carries the gene coding for poly(3-hydroxybutyrate) (PHB) depolymerase was cloned from the chromosomal DNA of Alcaligenes faecalis AE122 isolated from seawater. The open reading frame encoding the precursor of the PHB depolymerase was 1905 base pairs (bp) long, corresponding to a protein of 635 amino acid residues (Mr=65 208). The promoter site, which could be recognized by Escherichia coli RNA polymerase, was upstream from the gene, and the sequence adhering to the ribosome-binding sequence was found in front of the gene. The deduced amino acid sequence agreed with the N-terminal amino acid sequence of the purified PHB depolymerase from amino acid 28 onwards. Analysis of the deduced amino acid sequence revealed the domain structure of the protein; a signal peptide of 27 amino acids long was followed by a catalytic domain of about 400 amino acids, a fibronectin type III module sequence, and a putative substrate binding domain. The molecular mass (62 526) of the mature protein deduced from the nucleotide sequence was significantly lower than the value (95 kDa) estimated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but coincided well with the value (62 426) estimated from matrix-assisted laser desorption ionization mass spectra. By comparison of the primary structure with those of other PHB depolymerases, the substrate binding domain was found to consist of two domains, PHB-specific and poly(3-hydroxyvalerate)-specific ones, connected by a linker region. The PHB depolymerase gene was expressed in Escherichia coli under the control of the tac promoter. The enzyme expressed in E. coli was purified from culture broth and showed the same catalytic properties as the enzyme from A. faecalis.

This article was published in Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression and referenced in Journal of Pollution Effects & Control

Relevant Expert PPTs

Relevant Speaker PPTs

Recommended Conferences

Relevant Topics

Peer Reviewed Journals
 
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
International Conferences 2017-18
 
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

Agri, Food, Aqua and Veterinary Science Journals

Dr. Krish

agrifoodaquavet@omicsonline.com

1-702-714-7001 Extn: 9040

Clinical and Biochemistry Journals

Datta A

clinical_biochem@omicsonline.com

1-702-714-7001Extn: 9037

Business & Management Journals

Ronald

business@omicsonline.com

1-702-714-7001Extn: 9042

Chemical Engineering and Chemistry Journals

Gabriel Shaw

chemicaleng_chemistry@omicsonline.com

1-702-714-7001 Extn: 9040

Earth & Environmental Sciences

Katie Wilson

environmentalsci@omicsonline.com

1-702-714-7001Extn: 9042

Engineering Journals

James Franklin

engineering@omicsonline.com

1-702-714-7001Extn: 9042

General Science and Health care Journals

Andrea Jason

generalsci_healthcare@omicsonline.com

1-702-714-7001Extn: 9043

Genetics and Molecular Biology Journals

Anna Melissa

genetics_molbio@omicsonline.com

1-702-714-7001 Extn: 9006

Immunology & Microbiology Journals

David Gorantl

immuno_microbio@omicsonline.com

1-702-714-7001Extn: 9014

Informatics Journals

Stephanie Skinner

omics@omicsonline.com

1-702-714-7001Extn: 9039

Material Sciences Journals

Rachle Green

materialsci@omicsonline.com

1-702-714-7001Extn: 9039

Mathematics and Physics Journals

Jim Willison

mathematics_physics@omicsonline.com

1-702-714-7001 Extn: 9042

Medical Journals

Nimmi Anna

medical@omicsonline.com

1-702-714-7001 Extn: 9038

Neuroscience & Psychology Journals

Nathan T

neuro_psychology@omicsonline.com

1-702-714-7001Extn: 9041

Pharmaceutical Sciences Journals

John Behannon

pharma@omicsonline.com

1-702-714-7001Extn: 9007

Social & Political Science Journals

Steve Harry

social_politicalsci@omicsonline.com

1-702-714-7001 Extn: 9042

 
© 2008-2017 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version
adwords