Multi-copy expression and fed-batch production of Rhodotorula araucariae epoxide hydrolase in Yarrowia lipolytica

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dc.contributor.authorMaharajh D.
dc.contributor.authorRoth R.
dc.contributor.authorLalloo R.
dc.contributor.authorSimpson C.
dc.contributor.authorMitra R.
dc.contributor.authorGorgens J.
dc.contributor.authorRamchuran S.
dc.date.accessioned2011-05-15T15:56:36Z
dc.date.available2011-05-15T15:56:36Z
dc.date.issued2008
dc.description.abstractEpoxide hydrolases (EHs) of fungal origin have the ability to catalyze the enantioselective hydrolysis of epoxides to their corresponding diols. However, wild type fungal EHs are limited in substrate range and enantioselectivity. Additionally, the production of fungal epoxide hydrolase (EH) by wild-type strains is typically very low. In the present study, the EH-encoding gene from Rhodotorula araucariae was functionally expressed in Yarrowia lipolytica, under the control of a growth phase inducible hp4d promoter, in a multi-copy expression cassette. The transformation experiments yielded a positive transformant, with a final EH activity of 220 U/g dw in shake-flask cultures. Evaluation of this transformant in batch fermentations resulted in ∼ 7-fold improvement in EH activity over the flask scale. Different constant specific feed rates were tested in fed-batch fermentations, resulting in an EH activity of 1,750 U/g dw at a specific feed rate of ∼ 0.1 g/g/h, in comparison to enzyme production levels of 0.3 U/g dw for the wild type R. araucariae and 52 U/g dw for an Escherichia coli recombinant strain expressing the same gene. The expression of EH in Y. lipolytica using a multi-copy cassette demonstrates potential for commercial application. © 2008 Springer-Verlag.
dc.description.versionArticle
dc.identifier.citationApplied Microbiology and Biotechnology
dc.identifier.citation79
dc.identifier.citation2
dc.identifier.issn1757598
dc.identifier.other10.1007/s00253-008-1420-7
dc.identifier.urihttp://hdl.handle.net/10019.1/9938
dc.subjectCell culture
dc.subjectEnantioselectivity
dc.subjectEnzyme activity
dc.subjectFermentation
dc.subjectHydrolysis
dc.subjectEnzyme production
dc.subjectEpoxide hydrolases
dc.subjectFed-batch fermentation
dc.subjectRhodotorula araucariae
dc.subjectYarrowia lipolytica
dc.subjectFungi
dc.subjectepoxide
dc.subjectepoxide hydrolase
dc.subjectenzyme activity
dc.subjectgene expression
dc.subjecthydrolysis
dc.subjectyeast
dc.subjectarticle
dc.subjectbacterial strain
dc.subjectenantioselectivity
dc.subjectenzyme activity
dc.subjectenzyme mechanism
dc.subjectEscherichia coli
dc.subjectfed batch fermentation
dc.subjectfungal gene
dc.subjectfungal strain
dc.subjectfungus
dc.subjectgene cassette
dc.subjectgene expression
dc.subjecthp4d gene
dc.subjecthydrolysis
dc.subjectnonhuman
dc.subjectnucleotide sequence
dc.subjectpromoter region
dc.subjectRhodotorula araucariae
dc.subjectwild type
dc.subjectYarrowia lipolytica
dc.subjectBioreactors
dc.subjectBiotechnology
dc.subjectEpoxide Hydrolases
dc.subjectEpoxy Compounds
dc.subjectFermentation
dc.subjectFungal Proteins
dc.subjectGene Expression
dc.subjectRecombinant Fusion Proteins
dc.subjectRhodotorula
dc.subjectYarrowia
dc.subjectEscherichia coli
dc.subjectRhodotorula araucariae
dc.subjectYarrowia lipolytica
dc.titleMulti-copy expression and fed-batch production of Rhodotorula araucariae epoxide hydrolase in Yarrowia lipolytica
dc.typeArticle
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