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Molecular Cloning and Expression of a Putative Oleate Hydratase Isozyme From Nocardia Cholesterolicum NRRL 5767
Author(s) -
Huang JenqKuen,
Alhmadi Hekmat B,
Vanderway David R,
Hoerner Cole A,
Wen Lisa
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.838.1
Subject(s) - cloning (programming) , biochemistry , recombinant dna , enzyme , gene , isozyme , molecular cloning , oleic acid , clone (java method) , chemistry , microbiology and biotechnology , biology , gene expression , computer science , programming language
Nocardia Cholesterolicum NRRL 5767 is well‐known for the ability to transform about 95% of added oleic acid to value‐added product, namely 10‐hydroxystearic acid (10‐HSA). Oleate hydratase is the enzyme catalyzes this conversion. The objective of this research was to clone, overexpress, and characterize oleate hydratase. Nocardia Cholesterolicum NRRL 5767 genome contains two annotated oleate hydratase. Here, we report cloning of an annotated oleate hydratase gene from the N. cholestericum genomic DNA by PCR. The gene was inserted into pET28a. Expression of functional recombinant oleate hydratase was achieved by IPTG induction. The ability to produce large quantities of the protein will permit characterization and evaluation of its suitability for use in immobilized enzyme for the production of 10‐HSA. Support or Funding Information Western Illinois University Foundation and the Department of Chemistry.