Structure‐Based Mechanism of Oleate Hydratase from <i>Elizabethkingia meningoseptica</i>

Engleder, Matthias; Pavkov‐Keller, Tea; Emmerstorfer, Anita; Hromic, Altijana; Schrempf, Sabine; Steinkellner, Georg; Wriessnegger, Tamara; Leitner, Erich; Strohmeier, Gernot A.; Kaluzna, Iwona; Mink, Daniel; Schürmann, Martin; Wallner, Silvia; Macheroux, Peter; Gruber, Karl; Pichler, Harald

doi:10.1002/cbic.201500269

Cited by 69 publications

(183 citation statements)

References 21 publications

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“…All currently known FAHs share a conserved N-terminal nucleotide binding motif for non-covalent attachment of the essential flavin adenine dinucleotide (FAD) cofactor despite their high overall sequence diversity (Wierenga et al 1986; Kleiger and Eisenberg 2002). Therefore, all FAHs characterised so far are flavin-dependent proteins (Volkov et al 2010; Joo et al 2012a; Engleder et al 2015; Hirata et al 2015; Kang et al 2017). Since the redox (reduction/oxidation) state of FAD does not change during substrate conversion, FAHs are belonging to the approx.…”

Section: Enzymes For Hydration and Their Propertiesmentioning

confidence: 99%

On the current role of hydratases in biocatalysis

Engleder

Pichler

2018

Appl Microbiol Biotechnol

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Water addition to carbon-carbon double bonds provides access to value-added products from inexpensive organic feedstock. This interesting but relatively little-studied reaction is catalysed by hydratases in a highly regio- and enantiospecific fashion with excellent atom economy. Considering that asymmetric hydration of (non-activated) carbon-carbon double bonds is virtually impossible with current organic chemistry, enzymatic hydration reactions are highly attractive for industrial applications. Hydratases have been known for several decades but their biocatalytic potential has only been explored over the past 15 years. As a result, a considerable amount of information on this enzyme group has become available, enabling their development for practical applications. This review focuses on hydratases catalysing water addition to non-activated carbon-carbon double bonds, and examines hydratases from a biochemical, structural and mechanistic angle. Current challenges and opportunities in hydration biocatalysis are discussed, and, ultimately, their potential for organic synthesis is highlighted.

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Section: Enzymes For Hydration and Their Propertiesmentioning

confidence: 99%

On the current role of hydratases in biocatalysis

Engleder

Pichler

2018

Appl Microbiol Biotechnol

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“…The reaction mechanism of OhyA from E. meningoseptica proposed that FAD was involved in the correct localization of the substrate and amino acids in the active site of OhyA (7). In the present study, the cofactor effect on the activities of holo-OhyAs and role of the residue of FAD-binding motif on the enzyme stability were investigated.…”

Section: Discussionmentioning

confidence: 89%

“…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.…”

mentioning

confidence: 99%

“…All OhyAs contain flavin adenine dinucleotide (FAD)-binding motifs. When FAD was not bound to OhyA, its catalytic activity was abolished (12) or significantly reduced (7). Thus, OhyAs are FAD-dependent enzymes belonging to the myosin cross-reactive antigen (MCRA) protein family, and they are widely distributed in bacteria, especially in pathogenic and intestinal bacteria.…”

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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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“…LAH is a structural homodimer and possesses a long substrate binding channel for fatty acids with an N-terminal lid domain in each protomer. In 2015, Engleder et al, were able to resolve the holo structure of the oleate hydratase from Elizabethkingia meningoseptica with its FAD cofactor (pdb: 4UIR) [129].…”

Section: Fatty Acid Double Bond Hydratasesmentioning

confidence: 99%

Engineering and application of enzymes for lipid modification, an update

Zorn

Oroz‐Guinea

Brundiek³

et al. 2016

Progress in Lipid Research

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This review first provides a brief introduction into the most important tools and strategies for protein engineering (i.e. directed evolution and rational protein design combined with high-throughput screening methods) followed by examples from literature, in which enzymes have been optimized for biocatalytic applications. This covers engineered lipases with altered fatty acid chain length selectivity, fatty acid specificity and improved performance in esterification reactions. Furthermore, recent achievements reported for phospholipases, lipoxygenases, P450 monooxygenases, decarboxylating enzymes, fatty acid hydratases and the use of enzymes in cascade reactions are treated.

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Structure‐Based Mechanism of Oleate Hydratase from Elizabethkingia meningoseptica

Cited by 69 publications

References 21 publications

On the current role of hydratases in biocatalysis

On the current role of hydratases in biocatalysis

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

Engineering and application of enzymes for lipid modification, an update

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