Abstract
Soapstock is a by‐product of edible oil refining process with low cash value. It is rich in oleic and linoleic acids which can be converted to value‐added compounds by certain microorganisms. Nocardia cholesterolicum NRRL 5767 (N. cholesterolicum NRRL 5767) is a well‐known microbe that can transform oleic acid to 10‐hydroxyoctadecanoic acid (10‐HOOD, 85%) and 10‐oxo‐octadecanoic (10‐OXOD, 10%). The 10‐OXOD contaminant complicates downstream purification of 10‐OHOD. The conversion reactions are catalyzed by oleate dehydrogenase and secondary alcohol dehydrogenase. The long‐term goal is strain improvement of N. cholesterolicum NRRL 5767 by blocking 2‐ADH to create an industrially useful microbe for the production of 10‐OHOD from oleic acid. M. luteus was used for 2‐ADH purification due to its lack of oleate hydratase which hampered the purification of 2‐ADH from N. cholesterolicum NRRL 5767.The 2‐ADH from M. luteus WIUJH20 has been purified to homogeneity and its N‐terminal sequence determined. Using BLAST search tool, the 2‐ADH gene has been identified as belonging to the 3‐hydroxyacyl CoA dehydrogenase superfamily. The gene has been amplified by PCR and cloned into pGEX2T. The recombinant 2‐ADH protein has been overexpressed in E. coli, purified, and its Km and Vmax determined.Supported in part by grants from USDA CSREES 04‐35504‐14712 and 02‐015470.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
