Production of 10-hydroxystearic acid from oleic acid and olive oil hydrolyzate by an oleate hydratase from Lysinibacillus fusiformis

Abstract

A recombinant enzyme from Lysinibacillus fusiformis was expressed, purified, and identified as an oleate hydratase because the hydration activity of the enzyme was the highest for oleic acid (with a k (cat) of 850 min(-1) and a K (m) of 540 μM), followed by palmitoleic acid, γ-linolenic acid, linoleic acid, myristoleic acid, and α-linolenic acid. The optimal reaction conditions for the enzymatic production of 10-hydroxystearic acid were pH 6.5, 35 °C, 4% (v/v) ethanol, 2,500 U ml(-1) (8.3 mg ml(-1)) of enzyme, and 40 g l(-1) oleic acid. Under these conditions, 40 g l(-1) (142 mM) oleic acid was converted into 40 g l(-1) (133 mM) 10-hydroxystearic acid for 150 min, with a molar yield of 94% and a productivity of 16 g l(-1) h(-1), and olive oil hydrolyzate containing 40 g l(-1) oleic acid was converted into 40 g l(-1) 10-hydroxystearic acid for 300 min, with a productivity of 8 g l(-1) h(-1).

In vitro synthesis of 9,10-dihydroxyhexadecanoic acid using recombinant Escherichia coli

“…Different types of hydroxy fatty acid production has been reported with the help of microbes. Production of 10-hydroxystearic acid has been achieved by whole cell conversions using wild-type [19–21] and recombinant microorganisms [22]. 10-Hydroxystearic acid is produced from oleic acid using whole cells of recombinant E. coli expressing the oleate hydratase gene of Stenotrophomonas maltophilia [22].…”

Production of 13S-hydroxy-9(Z)-octadecenoic acid from linoleic acid by whole recombinant cells expressing linoleate 13-hydratase from Lactobacillus acidophilus

“…The solvent was removed from the extract using a rotary evaporator. The obtained fatty acids were silylated with a 3:1 mixture of pyridine and N-methyl-N-(trimethylsilyl)trifluoroacetamide (Kim et al, 2012). The silylated fatty acids in the organic phase were analyzed using a gas chromatography (GC) (6890N, Agilent Santa Clara, CA, USA) equipped with a flame ionization detector and a Supelco SPB-1 capillary column (Belafonte, PA, USA).…”

Engineering and application of enzymes for lipid modification, an update

“…Meanwhile several other ∆9 hydratases have been recombinantly expressed as for instance the oleate hydratases from Lysinibacillus fusiformis [130], Stenotrophomonas maltophilia [131] or from Macrococcus caseolyticus [132]. These oleate hydratases have in common, that they do not at all or only to a minor extent hydrate ∆12 double bonds in linoleic acid.…”