Enantioselective hydrolysis of unbranched aliphatic 1,2-epoxides by Rhodotorula glutinis

Weijers, C.A.G.M.; Botes, Adriana L.; Dyk, M. S. Van; Bont, J.A.M. de

doi:10.1016/s0957-4166(97)00639-3

Cited by 62 publications

(15 citation statements)

References 12 publications

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“…An increase of activity toward unbranched 1,2-epoxyalkanes with increasing lengths of the side group has also been found with epoxide hydrolases from R. glutinis and different Rhodothorula and Trichosporon spp. (15,48). The epoxide hydrolase from N. punctiforme (Npueh1) was active not only with all terminal epoxides that were tested but also with the 1,2-disubstituted epoxides trans-stilbene oxide (compound 7) and cyclohexene oxide (compound 23) (Tables 4 and 5).…”

Section: Vol 72 2006 Novel Epoxide Hydrolases From Genomic Databasementioning

confidence: 99%

Diversity and Biocatalytic Potential of Epoxide Hydrolases Identified by Genome Analysis

Loo

Kingma

Arand

et al. 2006

Appl Environ Microbiol

109

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Epoxide hydrolases play an important role in the biodegradation of organic compounds and are potentially useful in enantioselective biocatalysis. An analysis of various genomic databases revealed that about 20% of sequenced organisms contain one or more putative epoxide hydrolase genes. They were found in all domains of life, and many fungi and actinobacteria contain several putative epoxide hydrolase-encoding genes. Multiple sequence alignments of epoxide hydrolases with other known and putative ␣/␤-hydrolase fold enzymes that possess a nucleophilic aspartate revealed that these enzymes can be classified into eight phylogenetic groups that all contain putative epoxide hydrolases. To determine their catalytic activities, 10 putative bacterial epoxide hydrolase genes and 2 known bacterial epoxide hydrolase genes were cloned and overexpressed in Escherichia coli. The production of active enzyme was strongly improved by fusion to the maltose binding protein (MalE), which prevented inclusion body formation and facilitated protein purification. Eight of the 12 fusion proteins were active toward one or more of the 21 epoxides that were tested, and they converted both terminal and nonterminal epoxides. Four of the new epoxide hydrolases showed an uncommon enantiopreference for meso-epoxides and/or terminal aromatic epoxides, which made them suitable for the production of enantiopure (S,S)-diols and (R)-epoxides. The results show that the expression of epoxide hydrolase genes that are detected by analyses of genomic databases is a useful strategy for obtaining new biocatalysts.Enantiopure epoxides and vicinal diols are valuable intermediates in the synthesis of a number of pharmaceutical compounds. Epoxide hydrolases (EC 3.3.2.3) catalyze the conversion of epoxides to the corresponding diols. If they are enantioselective, they can be used to produce enantiopure epoxides by means of kinetic resolution (5). In the past, when only epoxide hydrolases from mammalian sources were known (12), the use of epoxide hydrolases in biocatalysis was hampered by their poor availability and insufficient catalytic performance, such as a low turnover rate or poor enantioselectivity. The potential for biocatalytic application of epoxide hydrolases was significantly increased with the discovery of microbial epoxide hydrolases (41), which are easier to produce in large quantities. The cloning and overexpression of several enantioselective epoxide hydrolases, e.g., from Agrobacterium radiobacter (35), Aspergillus niger (3), and potato plants (40), not only facilitated large-scale production of these enzymes but also made it possible to improve their biocatalytic properties by site-directed or random mutagenesis (34,36,43).Since many microbial genome sequences are available in the public domain, it is useful to screen these databases for genes that might encode new enzymes with interesting properties.Novel epoxide hydrolases can be identified by performing a BLAST search of the genomic databases, using amino acid sequences of known epoxide hyd...

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Section: Vol 72 2006 Novel Epoxide Hydrolases From Genomic Databasementioning

confidence: 99%

Diversity and Biocatalytic Potential of Epoxide Hydrolases Identified by Genome Analysis

Loo

Kingma

Arand

et al. 2006

Appl Environ Microbiol

109

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“…Twenty‐four commercially available aliphatic and aromatic epoxides ( 7 – 12 , Scheme ) were assayed with epoxide hydrolases from Aspergillus niger 19 and Rhodotorula glutinis. 20 The reactivity pattern for the substrate series was recorded with both enzymes under their optimal operating conditions (Figure 4). This is the first generally applicable chromogenic assay for epoxide hydrolases 21…”

Section: Methodsmentioning

confidence: 99%

The Adrenaline Test for Enzymes

Wahler¹,

Reymond²

2002

Angew. Chem.

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“…[18] Twenty-four commercially available aliphatic and aromatic epoxides (7 ± 12, Scheme 2) were assayed with epoxide hydrolases from Aspergillus niger [19] and Rhodotorula glutinis. [20] The reactivity pattern for the substrate series was recorded with both enzymes under their optimal operating conditions ( Figure 4). This is the first generally applicable chromogenic assay for epoxide hydrolases.…”

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confidence: 99%