Production of 10-hydroxystearic acid from oleic acid by whole cells of recombinant Escherichia coli containing oleate hydratase from Stenotrophomonas maltophilia

Joo, Young-Chul; Seo, Eun-Sun; Kim, Yeong-Su; Kim, Kyoung-Rok; Park, Jin Byung; Oh, Deok‐Kun

doi:10.1016/j.jbiotec.2012.01.002

Cited by 83 publications

(64 citation statements)

References 24 publications

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“…Cytochrome P450s, for example, are well known for their ability to break a C-H σ bond of fatty acid alkyl chain and attach a hydroxyl group to the carbon (Whitehouse et al 2012;Budde et al 2004;Lu et al 2010). Hydratases break a CC π bond in the middle of the alkyl chain and attach a hydroxyl group at one of the carbons (Joo et al 2012). Laccases convert unsaturated fatty acids into epoxy or hydroxy fatty acids (Virk et al 2012).…”

Section: Introductionmentioning

confidence: 99%

fadD deletion and fadL overexpression in Escherichia coli increase hydroxy long-chain fatty acid productivity

Bae

Park

Jung

et al. 2014

Appl Microbiol Biotechnol

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A major problem of long-chain fatty acid (LCFA) hydroxylation using Escherichia coli is that FadD (long-chain fatty acyl-CoA synthetase), which is necessary for exogenous LCFA transport, also initiates cellular consumption of LCFA. In this study, an effective method to prevent the cellular consumption of LCFA without impairing its transport is proposed. The main idea is that a heterologous enzyme which consumes LCFA can replace FadD in LCFA transport. For the model heterologous enzyme, CYP153A from Marinobacter aquaeolei, which converts palmitic acid into ω-hydroxy palmitic acid, was expressed in E. coli. When fadD was deleted from an E. coli strain, CYP153A indeed maintained the ability to transport LCFA. A disadvantage of fadD deletion mutant is the fact that FadD deficiency downregulates the transcription of fadL (outer membrane LCFA transporter) via FadR (fatty acid metabolism regulator protein), was solved by fadL overexpression from a plasmid. In addition, the overexpression of fadL was able to offset catabolite repression on fadL, allowing glucose to be used as the primary carbon source. In conclusion, the strain with fadD deletion and fadL overexpression showed 5.5-fold increase in productivity compared to the wild-type strain, converting 2.6 g/L (10.0 mM) of palmitic acid into 2.4 g/L (8.8 mM) of ω-hydroxy palmitic acid in a shake flask. This simple genetic manipulation can be applied to any LCFA hydroxylation using E. coli.

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Section: Introductionmentioning

confidence: 99%

fadD deletion and fadL overexpression in Escherichia coli increase hydroxy long-chain fatty acid productivity

Bae

Park

Jung

et al. 2014

Appl Microbiol Biotechnol

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“…WP_017356052), and the synthesized gene was cloned and expressed in the Escherichia coli strain ER2566. The gene encoding the original OhyA of S. maltophilia was 1,770 bp (13). The enzyme was then expressed and purified as a soluble protein from the supernatant of lysed cells using HisTrap affinity chromatography.…”

Section: Resultsmentioning

confidence: 99%

“…The biochemical properties of OhyAs from other bacteria, including S. pyogenes, M. caseolyticus (12), L. fusiformis (11), S. maltophilia (13), and B. breve (8,9), were subsequently characterized. These OhyAs showed low activity toward PUFAs.…”

Section: Discussionmentioning

confidence: 99%

“…The biochemical properties of OhyAs from Elizabethkingia meningoseptica (6,7), Bifidobacterium breve (8,9), Streptococcus pyogenes (10), Lysinibacillus fusiformis (11), Macrococcus caseolyticus (12), Stenotrophomonas maltophilia (13), and Stenotrophomonas nitritireducens (14) have been characterized to date. All OhyAs contain flavin adenine dinucleotide (FAD)-binding motifs.…”

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confidence: 99%

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

Kang

Seo

Shin

et al. 2017

Appl Environ Microbiol

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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 -hydroxycarboxylic and ␣, -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, ␣-linolenic acid, and ␥-linolenic acid, were greater than those of OhyA1. Moreover, the specific activity of S. maltophilia OhyA2 toward unsaturated fatty acids, with the exception of ␥-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, ␣-linolenic acid, and ␥-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. KEYWORDS Stenotrophomonas maltophilia, enzyme characterization, oleate hydrataseO leate hydratase (OhyA) converts unsaturated fatty acids to 10-hydroxy fatty acids, which are then converted to ␥-lactones by the yeasts Candida boidinii and Waltomyces lipofer (1, 2). ␥-Lactones are flavor compounds used in food and cosmetics (3). Multistep enzymatic reactions involving OhyA, alcohol dehydrogenase, Baeyer-Villiger monooxygenases, and esterase convert unsaturated fatty acids to -hydroxycarboxylic and ␣, -dicarboxylic acids (4), which are used in the preparation of resins, nylons, plastics, and lubricants (5). An effective OhyA is necessary for the efficient production of ␥-lactones, -hydroxycarboxylic acids, and ␣, -dicarboxylic acids.

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“…Rosberg-Cody et al, 2011), Lysinibacillus fusiformis(Kim et al, 2012;Seo et al, 2013), Stenotrophomonas maltophilia(Jeon et al, 2012;Joo et al, 2012b), Macrococcus caseolyticus(Joo et al, 2012a), Lactobacillus rhamnosus…”

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confidence: 99%

Stereochemistry of enzymatic water addition to C = C bonds

Chen

Otten

Hanefeld

2015

Biotechnology Advances

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Production of 10-hydroxystearic acid from oleic acid by whole cells of recombinant Escherichia coli containing oleate hydratase from Stenotrophomonas maltophilia

Cited by 83 publications

References 24 publications

fadD deletion and fadL overexpression in Escherichia coli increase hydroxy long-chain fatty acid productivity

fadD deletion and fadL overexpression in Escherichia coli increase hydroxy long-chain fatty acid productivity

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

Stereochemistry of enzymatic water addition to C = C bonds

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