Efficient synthesis of optically pure alcohols by asymmetric hydrogen-transfer biocatalysis: application of engineered enzymes in a 2-propanol–water medium

Itoh, Nobuhide; Isotani, Kentaro; Nakamura, Masaru; Inoue, Kousuke; Isogai, Yasuhiro; Makino, Yoshihide

doi:10.1007/s00253-011-3447-4

Cited by 41 publications

(41 citation statements)

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“…Previously, we reported an efficient method for producing both enantiomers of chiral alcohols by asymmetric hydrogentransfer bioreduction of ketones in a 2-propanol (IPA)-water medium using E. coli biocatalysts expressing a mutated form of phenylacetaldehyde reductase (PAR) (22,23) and Leifsonia ADH (LSADH) (24,25). However, PAR and LSADH do not fully possess the required substrate specificity or stereospecificity; for example, LSADH does not accept methyl benzoylformate, 2-acetylpyridine, or 3-quinuclidinone as a substrate (26,27).…”

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confidence: 99%

Efficient PCR-Based Amplification of Diverse Alcohol Dehydrogenase Genes from Metagenomes for Improving Biocatalysis: Screening of Gene-Specific Amplicons from Metagenomes

Kariya

Kurokawa

2014

Appl Environ Microbiol

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Screening of gene-specific amplicons from metagenomes (S-GAM) has tremendous biotechnological potential. We used this approach to isolate alcohol dehydrogenase (adh) genes from metagenomes based on the Leifsonia species adh gene (lsadh), the enzyme product of which can produce various chiral alcohols. A primer combination was synthesized by reference to homologs of lsadh, and PCR was used to amplify nearly full-length adh genes from metagenomic DNAs. All adh preparations were fused with lsadh at the terminal region and used to construct Escherichia coli plasmid libraries. Of the approximately 2,000 colonies obtained, 1,200 clones were identified as adh positive (ϳ60%). Finally, 40 adh genes, Hladh-001 to Hladh-040 (for homologous Leifsonia adh), were identified from 223 clones with high efficiency, which were randomly sequenced from the 1,200 clones. The Hladh genes obtained via this approach encoded a wide variety of amino acid sequences (8 to 99%). After screening, the enzymes obtained (HLADH-012 and HLADH-021) were confirmed to be superior to LSADH in some respects for the production of antiPrelog chiral alcohols. Metagenomics is an emerging and powerful tool for the isolation of genes, the enzyme products of which have industrial applications (1-6). Screening of metagenomic libraries to find novel enzymes or enzymes homologous to those described previously has been used successfully to isolate lipases (7, 8), amylases (9), amidases (10), oxidoreductases (11), dehydratases (12), cytochrome P450 (13), styrene monooxygenase (14), and other enzymes (1-6). However, most previous approaches featured metagenomic DNA extraction and Escherichia coli library construction, followed by sequence-or function/molecule-based screens of the library. Such approaches are very time-consuming and inefficient, especially in terms of detection; much of the DNA sequenced and analyzed is irrelevant, and target genes may be expressed ambiguously in E. coli host cells. PCR amplification of truncated genes from metagenomes would facilitate the identification of genes encoding superior enzymes and yield homologous gene sets that could be used for DNA shuffling (15). Although previous studies based on PCR-mediated methods that utilize primers designed from inner conserved sequences have been conducted for biocatalysts, including lipase (8), cytochrome P450 (13), 2,5-diketo-D-gluconic acid reductase (16), Pseudomonas alcohol dehydrogenase (ADH) (17), and other biocatalysts (3, 4, 6), the methods are not very efficient in many cases and often fail to produce complete functional genes.Enantioselective organic synthesis is useful for producing chiral synthones for the preparation of fine chemicals, including pharmaceuticals and agricultural chemicals. The asymmetric reduction of ketones is one of the most promising approaches, because no substrate is lost, in contrast to when racemic separation is performed. Chiral metal complexes, such as BINAP-Ru, have been used successfully as chemocatalysts in a number of cases of enantioselective synthe...

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Efficient PCR-Based Amplification of Diverse Alcohol Dehydrogenase Genes from Metagenomes for Improving Biocatalysis: Screening of Gene-Specific Amplicons from Metagenomes

Kariya

Kurokawa

2014

Appl Environ Microbiol

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“…S749. 162 The catalytic activity of artificial metalloenzymes 190 and 191 derived from bovine β-lactoglobuline for the ATH of aromatic ketones using sodium formate in water was reported. It depends not only on the metal (ruthenium or rhodium) but also on the chelating activity and the length of the lipidic chain.…”

Section: Enzymatic Reactionsmentioning

confidence: 99%

Catalytic asymmetric transfer hydrogenation of ketones: recent advances

Foubelo

Nájera

Yus

2015

Tetrahedron: Asymmetry

215

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Abstract-In this review article we will consider the main processes on asymmetric transfer hydrogenation (ATH) of ketones from 2008 up today. The most effective organometallic compounds (derived from Ru, Rh, Ir, Fe, Os, Ni, Co, and Re) and chiral ligands (derived from amino alcohols, diamines, sulphur-and phosphorous-containing compounds, as well as heterocyclic systems) will be shown paying special attention to functionalized substrates, tandem reactions, processes under nonconventional conditions, supported catalysts, dynamic kinetic resolutions (DKR), the use of water as a green solvent, theoretical and experimental studies on reaction mechanisms, enzymatic processes and finally applications to the total synthesis of biologically active organic molecules. _____________________________________________________________________________________

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“…After amino-acid sequence analysis, the recombinant PAR was expressed in Escherichia coli [51]. One PAR mutant (HAR-1) could asymmetrically reduce 195 g L −1 OPBE to (R)-HPBE with 99% ee and 89% isolation yield [52]. The shotgun library of OPBE [53].…”

Section: Selected Opb(e) Redutasesmentioning

confidence: 99%

Bioreductive preparation of ACE inhibitors precursor (R)-2-hydroxy-4-phenylbutanoate esters: Recent advances and future perspectives

2015

Bioresour. Bioprocess.

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Optically active (R)-2-hydroxy-4-phenylbutanoate esters ((R)-HPBE) are key precursors for the production of angiotension-converting enzyme (ACE) inhibitors, which are important prescriptive drugs for preventing the formation of angiotensin II and lowering the blood pressure. The biocatalytic asymmetric reduction of ethyl 2-oxo-4-phenylbutanoate (OPBE) to (R)-HPBE with carbonyl reductases has several advantageous attributes, including high enantioselectivity, mild reaction condition, high catalytic efficiency, and environmental benignity. An increasing number of OPBE reductases have been discovered owing to the drastic achievements in genomics, screening and evolution technologies, and process engineering. The potential of (R)-HPBE production process has also been intensively evaluated. This review covers recent progress on the bioreductive preparation of (R)-HPBE, especially on various screening approaches for the identification of OPBE reductases and their characterization.

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Efficient synthesis of optically pure alcohols by asymmetric hydrogen-transfer biocatalysis: application of engineered enzymes in a 2-propanol–water medium

Cited by 41 publications

References 31 publications

Efficient PCR-Based Amplification of Diverse Alcohol Dehydrogenase Genes from Metagenomes for Improving Biocatalysis: Screening of Gene-Specific Amplicons from Metagenomes

Efficient PCR-Based Amplification of Diverse Alcohol Dehydrogenase Genes from Metagenomes for Improving Biocatalysis: Screening of Gene-Specific Amplicons from Metagenomes

Catalytic asymmetric transfer hydrogenation of ketones: recent advances

Bioreductive preparation of ACE inhibitors precursor (R)-2-hydroxy-4-phenylbutanoate esters: Recent advances and future perspectives

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