{'fraction_non_alphanumeric': 0.020058997050147492, 'fraction_numerical': 0.01887905604719764, 'mean_word_length': 5.352059925093633, 'pattern_counts': {'":': 0, '<': 0, '<?xml version=': 0, '>': 0, 'https://': 0, 'lorem ipsum': 0, 'www.': 0, 'xml': 0}, 'pii_count': 0, 'substrings_counts': 0, 'word_list_counts': {'cursed_substrings.json': 0, 'profanity_word_list.json': 0, 'sexual_word_list.json': 0, 'zh_pornsignals.json': 0}}
Molecular Cloning and Expression of an Oleate Hydratase From Nocardia Cholesterolicum NRRL 5767

The production of household cooking oils in an industrial setting yields soapstock as a byproduct during the refining process. Soapstock contains a significant amount of oleic acid and due to soapstocks very low price, it serves as a cheap source of oleic acid. Oleate hydratase (Ohase) converts oleic acid to the industrially useful product 10(R)‐hydroxystearic acid (10‐HSA), in a regioselective manner. Classically, 10‐HSA is utilized to thicken grease, but it has potential to be used in the production of plastics, lubricants and other various industrially relevant materials. Barring the use of biocatalytic microorganisms containing Ohase, this reaction would take place under unfavorable conditions involving the use of high temperatures/pressures, strong acids or metal catalysts, and all of these methods lead to little regioselectivity in the final product. Therefore, the use of a biocatalyst that catalyzes the conversion of oleate hydratase to 10‐HSA is extremely desirable for industrial applications. The bacterium Nocardia Cholesterolicum species NRRL 5767 is able to convert 95% of oleic acid added into the bacterial culture to 10‐HSA. Ohase has never been isolated or characterized at either the protein level or DNA level from NRRL 5767. This work is the first report of molecular cloning of the Ohase gene from NRRL 5767 genomic DNA into the pET28a expression vector. Functional recombinant Ohase enzyme has been expressed and purified. Enzyme activity of Ohase was determined by the conversion of oleic acid to 10‐HSA or by spectrophotometrically through a coupling reaction.