Abstract
Different microbial strains are able to transform oleic acid (OA) into 10-hydroxystearic acid (10-HSA) by means of the catalytic activity of the enzymes oleate hydratase (EC 4.2.1.53). Lactobacillus rhamnosus ATCC 53103 performs this biotransformation with very high stereoselectivity, affording enantiopure (R)-10-HSA. In this work, we cloned, in Escherichia coli, the oleate hydratase present in the above-mentioned probiotic strain. Our study demonstrated that the obtained recombinant hydratase retains the catalytic properties of the Lactobacillus strain but that its activity was greatly affected by the expression procedure. According to our findings, we devised a reliable procedure for the hydration of oleic acid using a recombinant E. coli whole-cell catalyst. We established that the optimal reaction conditions were pH 6.6 at 28 °C in phosphate buffer, using glycerol and ethanol as co-solvents. According to our experimental protocol, the biocatalyst does not show significant substrate inhibition as the hydration reaction can be performed at high oleic acid concentration (up to 50 g/L).
Highlights
The enzymatic addition of water to the double bond of free fatty acids was first described, in 1962, by Wallen et al [1] in the course of a study on the ability of the Pseudomonas sp. strain 3266 to catalyze the conversion of oleic acid 1 (OA) into 10-hydroxystearic acid 4 (10-HSA) (Figure 1).Catalysts 2020, 10, 1122; doi:10.3390/catal10101122 www.mdpi.com/journal/catalystsCatalysts 2020, 10, x FOR PEER REVIEW Figure Figure1
We have previously demonstrated the versatility of the probiotic bacteria Lactobacillus rhamnosus of the formation of diols and triols
Oleate hydratase from Lactobacillus rhamnosus ATCC 53103 was selected due to the positive results obtained with this strain in converting OA into 10-HSA [6]
Summary
The enzymatic addition of water to the double bond of free fatty acids was first described, in 1962, by Wallen et al [1] in the course of a study on the ability of the Pseudomonas sp. Strain 3266 to catalyze the conversion of oleic acid 1 (OA) into 10-hydroxystearic acid 4 (10-HSA) (Figure 1). Catalysts 2020, 10, x FOR PEER REVIEW Figure Figure. 1. The most unsaturated fatty oleic acids: oleic (1), linoleic (2)(3)and (3). 1. The mostcommon common CC1818 unsaturated fatty acids:. (1), linoleic (2) and linolenic acidlinolenic and acid and their transformation into (R)-10-hydroxystearic acid (4), (S)-(12Z)-10-hydroxy-octadecenoic their transformation into (R)-10-hydroxystearic acid (4), (S)-(12Z)-10-hydroxy-octadecenoic acid (5). (a) acid (5)and and(S)-(12Z,15Z)-10-hydroxy-octadecadienoic (S)-(12Z,15Z)-10-hydroxy-octadecadienoic (6), respectively.
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