Oleate hydratase

C. To isolated double bonds (Example oleate hydratase)... 

Occlusion zones 29 Octahedral geometry 311 Octopus, intelligence of 24 Oils, composition of 380 Okadaic acid 545 Old yellow enzyme 783 Oleate hydratase 526, 688 Oleic acid 381 Olfactory responses 558 Oligomer(s)... 

The enzyme oleate hydratase (E.C. 4.2.1.53) from Pseudomonas catalyzes the elimination of water from (R)-lO-hydroxystearate or the addition of water to a number of free unsaturated fatty acids, yielding (R)-lO-hydroxy fatty acids. Substrates that have been identified include linoleic acid, oleic acid and palmitoleic acid, which are converted to the corresponding 10-hydroxy-fatty acids 42l... 

Huang, J.-K., C.T. Hou, and M.O. Bagby Purification and Characterization of Oleate Hydratase from Nocardia cholesterolicum NRRL 5767 Physical and Chemical Properties, 34th West Central States Biochemistry Conference, Ames, lA, Abstract p. 51 (1991). 

Many studies have focused on 10-hydroxystearic acid production from oleic acid, but production via recombinant cells has been attempted only recently [138]. A putative fatty acid hydratase from Stenotrophomonas maltophilia was cloned and expressed in E. coli. The recombinant enzyme showed the highest hydration activity for oleic acid among the fatty acids tested, indicating that the enzyme is an oleate hydratase. The optimal conditions for the production of 10-hydroxystearic acid were pH 6.5, 35 °C, 0.05% w/v Tween 40,10 gl cells, and 50gl oleic acid. Under these conditions, whole recombinant cells produced 49gl 10-hydroxystearic acid for 4h, with a conversion yield of 98%w/w and a volumetric productivity of 12.3 gl h , which were 18% and 2.5-fold higher than those of whole wild-type S. maltophilia cells, respectively. 

K.R., Park, J.B., and Oh, D.K. (2012) Production of 10-hydroxystearic acid from oleic acid by whole cells of recombinant Escherichia coli containing oleate hydratase from Stenotrophomonas maltophilia.). Biotechnol, 158, 17-23, and references therein. 

However, oleate hydratase (EC 4.2.1.53), an enzyme from the class of hydrolyases, can catalyse this reaction on oleate creating (R)-lO-hydrohystearate and vice versa. The enzyme and its appropriate gene were purified from Elizabethkingia meningoseptica and cloned and expressed in E. coli (Bevers et al. 2009). The problem is that this enzyme has a relatively narrow specificity and cannot catalyse a conversion from isobutanol to isobutene. 

Bevers LE et al (2009) Oleate hydratase catalyzes the hydration of a nonactivated carbon-carbon bond. J Bacteriol 191(15) 5010-2... 

Kim BN, Joo YC, Kim YS, Kim KR, Oh DK (2012) Production of 10-hydroxystearic acid fi om oleic acid and olive oil hydrolyzate by an oleate hydratase fi om Lysinibacillus fiisiformis. Appl Microbiol Biotechnol 95(4) 929-937. doi 10.1007/s00253-011-3805-2 Kongo JM (ed) (2013) Lactic acid bacteria—R D for food, health and livestock purposes, InTech, Rijeka, Croatia, doi 10.5772/2825 ISBN 978-953-51-0955-6 Klimin B, Mlinch T (1998) Microbiological process for producing gamma-decalactone. US patent No 5 849551A... 

Oleate is an abundant 18-carbon monounsaturated fatty acid with a cis double bond between C-9 and C-10 (denoted A ). In the first step of oxidation, oleate is converted to oleoyl-CoA and, like the saturated fatty acids, enters the mitochondrial matrix via the carnitine shuttle (Fig. 17-6). Oleoyl-CoA then undergoes three passes through the fatty acid oxidation cycle to yield three molecules of acetyl-CoA and the coenzyme A ester of a A, 12-carbon unsaturated fatty acid, cis-A -dodecenoyl-CoA (Fig. 17-9). This product cannot serve as a substrate for enoyl-CoA hydratase, which acts only on trans double bonds. The auxiliary enzyme A, A -enoyl-CoA isomerase isomerizes the ci5-A -enoyl-CoA to the fra/J5-A -enoyl-CoA, which is converted by enoyl-CoA hydratase into the corresponding L-/3-hydroxyacyl-CoA (fra/75-A -dodecenoyl-CoA). This intermediate is now acted upon by the remaining enzymes of /3 oxidation to yield acetyl-CoA and the coenzyme A ester of a 10-carbon saturated fatty acid, decanoyl-CoA. The latter undergoes four more passes through the pathway to yield five more molecules of acetyl-CoA. Altogether, nine acetyl-CoAs are produced from one molecule of the 18-carbon oleate. 

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