Biocatalytic study of novel oleate hydratases

[Display omitted] •Establishment of Hydratase Engineering Database comprising 2046 oleate hydratases.•Cloning and heterologous expression of nine selected oleate hydratase homologs.•Substrate specificities and hydration activities towards fatty acids were studied.•Activity towards novel truncated C1...

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Published inJournal of molecular catalysis. B, Enzymatic Vol. 133; pp. S243 - S249
Main Authors Schmid, Jens, Steiner, Lisa, Fademrecht, Silvia, Pleiss, Jürgen, Otte, Konrad B., Hauer, Bernhard
Format Journal Article
LanguageEnglish
Published Elsevier B.V 01.11.2016
Subjects
Alkenes
Biocatalysis
Database analysis
Hydration
Oleate hydratases
CC
Tt
Mp
Biocatalysis
Cg
Alkenes
Database analysis
Hydration
Gm
HFam(s)
Em
OAH(s)
Ht
HyED
Oleate hydratases
La
Db
Lj
gDNA
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Summary:[Display omitted] •Establishment of Hydratase Engineering Database comprising 2046 oleate hydratases.•Cloning and heterologous expression of nine selected oleate hydratase homologs.•Substrate specificities and hydration activities towards fatty acids were studied.•Activity towards novel truncated C11 fatty acid analog observed. The direct hydration of CC bonds to yield alcohols or the reverse dehydration is chemically challenging but highly sought after. Recently, oleate hydratases (OAHs) gained attention as biocatalytic alternatives capable of hydrating isolated, non-activated CC bonds. Their natural reaction is the conversion of oleic acid to (R)-10-hydroxystearic acid. In this work, we report the first comparative study of several OAHs. Therefore we established the Hydratase Engineering Database (HyED) comprising 2046 putative OAHs from eleven homologous families and selected nine homologs for cloning in E. coli. The heterologously expressed enzymes were evaluated concerning activity and substrate specificity. The enzymes have a broad substrate scope ranging from oleic acid (C18) to the novel synthetic substrate (Z)-undec-9-enoic acid (C11). The OAHs from Elizabethkingia meningoseptica and Chryseobacterium gleum showed the best expression, highest stability and broadest substrate scope, making them interesting candidates for directed evolution to engineer them for the application as general hydratase catalysts.
ISSN:1381-1177
1873-3158
DOI:10.1016/j.molcatb.2017.01.010