Jens Schmid , Lisa Steiner , Silvia Fademrecht, Jürgen Pleiss, Konrad B. Otte, Bernhard Hauer
{"title":"新型油酸水合酶的生物催化研究","authors":"Jens Schmid , Lisa Steiner , Silvia Fademrecht, Jürgen Pleiss, Konrad B. Otte, Bernhard Hauer","doi":"10.1016/j.molcatb.2017.01.010","DOIUrl":null,"url":null,"abstract":"<div><p>The direct hydration of C<img>C 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 C<img>C bonds. Their natural reaction is the conversion of oleic acid to (<em>R</em>)-10-hydroxystearic acid.</p><p>In this work, we report the first comparative study of several OAHs. Therefore we established the <em>Hydratase Engineering Database</em> (HyED) comprising 2046 putative OAHs from eleven homologous families and selected nine homologs for cloning in <em>E. coli</em>. 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 (<em>Z</em>)-undec-9-enoic acid (C11). The OAHs from <em>Elizabethkingia meningoseptica</em> and <em>Chryseobacterium gleum</em> 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.</p></div>","PeriodicalId":16416,"journal":{"name":"Journal of Molecular Catalysis B-enzymatic","volume":"133 ","pages":"Pages S243-S249"},"PeriodicalIF":0.0000,"publicationDate":"2016-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molcatb.2017.01.010","citationCount":"24","resultStr":"{\"title\":\"Biocatalytic study of novel oleate hydratases\",\"authors\":\"Jens Schmid , Lisa Steiner , Silvia Fademrecht, Jürgen Pleiss, Konrad B. Otte, Bernhard Hauer\",\"doi\":\"10.1016/j.molcatb.2017.01.010\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>The direct hydration of C<img>C 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 C<img>C bonds. Their natural reaction is the conversion of oleic acid to (<em>R</em>)-10-hydroxystearic acid.</p><p>In this work, we report the first comparative study of several OAHs. Therefore we established the <em>Hydratase Engineering Database</em> (HyED) comprising 2046 putative OAHs from eleven homologous families and selected nine homologs for cloning in <em>E. coli</em>. 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 (<em>Z</em>)-undec-9-enoic acid (C11). The OAHs from <em>Elizabethkingia meningoseptica</em> and <em>Chryseobacterium gleum</em> 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.</p></div>\",\"PeriodicalId\":16416,\"journal\":{\"name\":\"Journal of Molecular Catalysis B-enzymatic\",\"volume\":\"133 \",\"pages\":\"Pages S243-S249\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2016-11-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/j.molcatb.2017.01.010\",\"citationCount\":\"24\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Molecular Catalysis B-enzymatic\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1381117717300103\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"Chemical Engineering\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Molecular Catalysis B-enzymatic","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1381117717300103","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Chemical Engineering","Score":null,"Total":0}
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.
期刊介绍:
Journal of Molecular Catalysis B: Enzymatic is an international forum for researchers and product developers in the applications of whole-cell and cell-free enzymes as catalysts in organic synthesis. Emphasis is on mechanistic and synthetic aspects of the biocatalytic transformation.
Papers should report novel and significant advances in one or more of the following topics;
Applied and fundamental studies of enzymes used for biocatalysis;
Industrial applications of enzymatic processes, e.g. in fine chemical synthesis;
Chemo-, regio- and enantioselective transformations;
Screening for biocatalysts;
Integration of biocatalytic and chemical steps in organic syntheses;
Novel biocatalysts, e.g. enzymes from extremophiles and catalytic antibodies;
Enzyme immobilization and stabilization, particularly in non-conventional media;
Bioprocess engineering aspects, e.g. membrane bioreactors;
Improvement of catalytic performance of enzymes, e.g. by protein engineering or chemical modification;
Structural studies, including computer simulation, relating to substrate specificity and reaction selectivity;
Biomimetic studies related to enzymatic transformations.