Bioprocess engineering to produce 10-hydroxystearic acid from oleic acid by recombinant Escherichia coli expressing the oleate hydratase gene of Stenotrophomonas maltophilia

2012 ◽  
Vol 47 (6) ◽  
pp. 941-947 ◽  
Author(s):  
Eun-Yeong Jeon ◽  
Jung-Hoo Lee ◽  
Kyung-Mi Yang ◽  
Young-Chul Joo ◽  
Deok-Kun Oh ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Oleic Acid ◽  
Stenotrophomonas Maltophilia ◽  
Recombinant Escherichia Coli ◽  
Hydroxystearic Acid ◽  
Bioprocess Engineering ◽  
Oleate Hydratase

scholarly journals Comparison of Biochemical Properties of the Original and Newly Identified Oleate Hydratases from Stenotrophomonas maltophilia

2017 ◽  
Vol 83 (9) ◽  
Author(s):  
Woo-Ri Kang ◽  
Min-Ju Seo ◽  
Kyung-Chul Shin ◽  
Jin-Byung Park ◽  
Deok-Kun Oh
Keyword(s):  
Fatty Acids ◽  
Oleic Acid ◽  
Linoleic Acid ◽  
Linolenic Acid ◽  
Stenotrophomonas Maltophilia ◽  
Unsaturated Fatty Acids ◽  
Hydroxy Fatty Acids ◽  
Plant Oils ◽  
Content Type ◽  
Oleate Hydratase

ABSTRACT Oleate hydratases (OhyAs) catalyze the conversion of unsaturated fatty acids to 10-hydroxy fatty acids, which are used as precursors of important industrial compounds, including lactones and ω-hydroxycarboxylic and α,ω-dicarboxylic acids. The genes encoding OhyA and a putative fatty acid hydratase in Stenotrophomonas maltophilia were identified by genomic analysis. The putative fatty acid hydratase was purified and identified as an oleate hydratase (OhyA2) based on its substrate specificity. The activity of OhyA2 as a holoenzyme was not affected by adding cofactors, whereas the activity of the original oleate hydratase (OhyA1) showed an increase. Thus, all characterized OhyAs were categorized as either OhyA1 or OhyA2 based on the activities of holoenzymes upon adding cofactors, which were determined by the type of the fourth conserved amino acid of flavin adenine dinucleotide (FAD)-binding motif. The hydration activities of S. maltophilia OhyA2 toward unsaturated fatty acids, including oleic acid, palmitoleic acid, linoleic acid, α-linolenic acid, and γ-linolenic acid, were greater than those of OhyA1. Moreover, the specific activity of S. maltophilia OhyA2 toward unsaturated fatty acids, with the exception of γ-linolenic acid, was the highest among all reported OhyAs. IMPORTANCE All characterized OhyAs were categorized as OhyA1s or OhyA2s based on the different properties of the reported and newly identified holo-OhyAs in S. maltophilia upon the addition of cofactors. OhyA2s showed higher activities toward polyunsaturated fatty acids (PUFAs), including linoleic acid, α-linolenic acid, and γ-linolenic acid, than those of OhyA1s. This suggests that OhyA2s can be used more effectively to convert plant oils to 10-hydroxy fatty acids because plant oils contain not only oleic acid but also PUFAs. The hydration activity of the newly identified OhyA2 from S. maltophilia toward oleic acid was the highest among the activity levels reported so far. Therefore, this enzyme is an efficient biocatalyst for the conversion of plant oils to 10-hydroxy fatty acids, which can be further converted to important industrial materials.

scholarly journals Sustainable Biotransformation of Oleic Acid to 10-Hydroxystearic Acid by a Recombinant Oleate Hydratase from Lactococcus garvieae

Processes ◽  
2019 ◽  
Vol 7 (6) ◽  
pp. 326 ◽  
Author(s):  
Jing Zhang ◽  
Muhammad Bilal ◽  
Shuai Liu ◽  
Jiaheng Zhang ◽  
Hedong Lu ◽  
...  
Keyword(s):  
Oleic Acid ◽  
Fatty Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Practical Interest ◽  
Hydroxystearic Acid ◽  
Enzymatic Production ◽  
Lactococcus Garvieae ◽  
Sds Page ◽  
Oleate Hydratase

Enzymatic hydration of oleic acid into 10-hydroxystearic acid (10-HSA) represents a theme of substantial scientific and practical interest. In this study, a fatty acid hydratase (OHase) from Lactococcus garvieae was cloned and expressed in Escherichia coli. The recombinantly expressed enzyme was identified as oleate hydratase (EC 4.2.1.53) confirming its highest hydration activity for oleic acid. The optimally yielded enzyme fraction was purified and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). A solitary band on SDS-PAGE confirmed the molecular weight of 65 kDa. Gas chromatography-mass spectrometry (GC-MS) analysis scrutinized the silylated hydroxy fatty acid products acquired from the hydration of oleic acid by the oleate hydratase from L. garvieae. Optimal reaction conditions for the enzymatic production of 10-HSA from oleic acid using the purified oleate hydratase were pH 7.5, 30 °C, 105.49 U/mL enzyme solution and 30 g/L oleic acid. In the presence of activity stimulators, that is, magnesium (II) (Mg2+), the oleate hydratase activity was found to be greatly improved at 30 °C. In conclusion, the results revealed the potential efficacy of recombinant enzyme for the biotechnological conversion of oleic acid to 10-HSA acid with high efficiency. The results would be useful for the improved industrial-scale biosynthesis of 10-HSA via an economical and environmentally friendly bioprocess approach.

scholarly journals Oleate Hydratase Catalyzes the Hydration of a Nonactivated Carbon-Carbon Bond

2009 ◽  
Vol 191 (15) ◽  
pp. 5010-5012 ◽  
Author(s):  
Loes E. Bevers ◽  
Martijn W. H. Pinkse ◽  
Peter D. E. M. Verhaert ◽  
Wilfred R. Hagen
Keyword(s):  
Oleic Acid ◽  
Cell Extract ◽  
Hydroxystearic Acid ◽  
Carbon Bond ◽  
Isolation And Characterization ◽  
Elizabethkingia Meningoseptica ◽  
Carbon Carbon Bond ◽  
Oleate Hydratase

ABSTRACT The hydration of oleic acid into 10-hydroxystearic acid was originally described for a Pseudomonas cell extract almost half a century ago. In the intervening years, the enzyme has never been characterized in any detail. We report here the isolation and characterization of oleate hydratase (EC 4.2.1.53) from Elizabethkingia meningoseptica.

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