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Screening of two complementary collections of Saccharomyces cerevisiae to identify enzymes involved in stereo-selective reductions of specific carbonyl compounds: an alternative to protein purification.

Katz, Michael LU ; Hahn-Hägerdal, Bärbel LU and Gorwa-Grauslund, Marie-Francoise LU (2003) In Enzyme and Microbial Technology 33(2-3). p.163-172
Abstract
Two collections of Saccharomyces cerevisiae strains, having open reading frames (ORFs) either overexpressed or deleted, were screened to identify cytosolic NADPH-dependent reductases involved in stereo-selective reductions of specific carbonyl compounds. As model compounds diacetyl (diketone) and ethyl acetoacetate (3-oxo ester) were used. The reductases encoded by YBR149w, YDR368w, YMR226c and YOR120w were found to reduce diacetyl and YOL151w, YHR104w, YGL157w, YOR120w and YDR368w reduced ethyl acetoacetate.



We cloned YBR149w, YDR368w and YMR226c using a vector with the strong constitutive GPD promotor and investigated the encoded reductases with respect to reduction of diacetyl. YBR149w, YDR368w and YMR226c were shown... (More)
Two collections of Saccharomyces cerevisiae strains, having open reading frames (ORFs) either overexpressed or deleted, were screened to identify cytosolic NADPH-dependent reductases involved in stereo-selective reductions of specific carbonyl compounds. As model compounds diacetyl (diketone) and ethyl acetoacetate (3-oxo ester) were used. The reductases encoded by YBR149w, YDR368w, YMR226c and YOR120w were found to reduce diacetyl and YOL151w, YHR104w, YGL157w, YOR120w and YDR368w reduced ethyl acetoacetate.



We cloned YBR149w, YDR368w and YMR226c using a vector with the strong constitutive GPD promotor and investigated the encoded reductases with respect to reduction of diacetyl. YBR149w, YDR368w and YMR226c were shown to encode (S)-specific diacetyl reductases capable of further reducing racemic acetoin to butanediol. Furthermore, we demonstrated that the purified (S)-diacetyl reductase (EC 1.2.1.56) most likely is the Image-arabinose dehydrogenase encoded by YBR149w (Ara1p). (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Screening, Reductase, Yeast, NADPH, Deletion library, Overexpression library, Open reading frame, Diacetyl, Ethyl acetoacetate
in
Enzyme and Microbial Technology
volume
33
issue
2-3
pages
163 - 172
publisher
Elsevier
external identifiers
  • wos:000184588900004
  • scopus:0142186831
ISSN
0141-0229
DOI
language
English
LU publication?
yes
id
658ef72c-681a-490f-b983-f9973731cea0 (old id 132807)
date added to LUP
2007-06-28 14:07:31
date last changed
2018-05-29 09:21:06
@article{658ef72c-681a-490f-b983-f9973731cea0,
  abstract     = {Two collections of Saccharomyces cerevisiae strains, having open reading frames (ORFs) either overexpressed or deleted, were screened to identify cytosolic NADPH-dependent reductases involved in stereo-selective reductions of specific carbonyl compounds. As model compounds diacetyl (diketone) and ethyl acetoacetate (3-oxo ester) were used. The reductases encoded by YBR149w, YDR368w, YMR226c and YOR120w were found to reduce diacetyl and YOL151w, YHR104w, YGL157w, YOR120w and YDR368w reduced ethyl acetoacetate.<br/><br>
<br/><br>
We cloned YBR149w, YDR368w and YMR226c using a vector with the strong constitutive GPD promotor and investigated the encoded reductases with respect to reduction of diacetyl. YBR149w, YDR368w and YMR226c were shown to encode (S)-specific diacetyl reductases capable of further reducing racemic acetoin to butanediol. Furthermore, we demonstrated that the purified (S)-diacetyl reductase (EC 1.2.1.56) most likely is the Image-arabinose dehydrogenase encoded by YBR149w (Ara1p).},
  author       = {Katz, Michael and Hahn-Hägerdal, Bärbel and Gorwa-Grauslund, Marie-Francoise},
  issn         = {0141-0229},
  keyword      = {Screening,Reductase,Yeast,NADPH,Deletion library,Overexpression library,Open reading frame,Diacetyl,Ethyl acetoacetate},
  language     = {eng},
  number       = {2-3},
  pages        = {163--172},
  publisher    = {Elsevier},
  series       = {Enzyme and Microbial Technology},
  title        = {Screening of two complementary collections of Saccharomyces cerevisiae to identify enzymes involved in stereo-selective reductions of specific carbonyl compounds: an alternative to protein purification.},
  url          = {http://dx.doi.org/},
  volume       = {33},
  year         = {2003},
}