Characterization of Ypr1p from Saccharomyces cerevisiae as a 2‐methylbutyraldehyde reductase

Yeast - Tập 19 Số 12 - Trang 1087-1096 - 2002
Gordon Ford1, Elizabeth M. Ellis2,1
1Department of Pharmaceutical Sciences, University of Strathclyde, 204 George Street, Glasgow G1 1XW, UK
2Department of Bioscience, University of Strathclyde, 204 George Street, Glasgow G1 1XW, UK

Tóm tắt

AbstractThe metabolism of aldehydes and ketones in yeast is important for biosynthetic, catabolic and detoxication processes. Aldo‐keto reductases are a family of enzymes that are able to reduce aldehydes and ketones. The roles of individual aldo‐keto reductases in yeast has been difficult to determine because of overlapping substrate specificities of these enzymes. In this study, we have cloned, expressed and characterized the aldo‐keto reductase Ypr1p from the yeast Saccharomyces cerevisiae and we describe its substrate specificity. The enzyme displays high specific activity towards 2‐methylbutyraldehyde, as well as other aldehydes such as hexanal. It exhibits extremely low activity as a glycerol dehydrogenase. The enzyme functions over a wide pH range and uses NADPH as co‐factor. In comparison to other mammalian and yeast aldo‐keto reductases, Ypr1p has relatively high affinity for D,L‐glyceraldehyde (1.08 mM) and hexanal (0.39 mM), but relatively low affinity for 4‐nitrobenzaldehyde (1.07 mM). It displays higher specific activity for 2‐methylbutyraldehyde than does yeast alcohol dehydrogenase and has a Km for 2‐methyl butyraldehyde of 1.09 mM. The enzyme is expressed during growth on glucose, but its levels are rapidly induced by osmotic and oxidative stress. Yeast in which the YPR1 gene has been deleted possess 50% lower 2‐methylbutyraldehyde reductase activity than the wild‐type strain. This suggests that the enzyme may contribute to 2‐methyl butyraldehyde reduction in vivo. It may therefore play a role in isoleucine catabolism and fusel alcohol formation and may influence flavour formation by strains of brewing yeast. Copyright © 2002 John Wiley & Sons, Ltd.

Từ khóa


Tài liệu tham khảo

10.1016/0378-1119(91)90592-Y

10.1093/nar/21.14.3329

10.1016/S0003-9861(51)80013-4

Bradford MM, 1979, A rapid and sensitive method for the quantification of protein utilizing the principle of protein‐dye binding, Anal Biochem, 72, 248, 10.1016/0003-2697(76)90527-3

10.1074/jbc.275.15.10937

10.1042/bj3120535

10.1073/pnas.90.21.10350

10.1016/S0009-2797(00)00259-3

Ganzhorn AJ, 1987, Kinetic characterization of yeast alcohol dehydrogenases: amino acid residue 294 and substrate specificity, J Biol Chem, 262, 2754, 10.1016/S0021-9258(18)61419-X

Gietz RD, 1995, Transforming yeast with DNA, Methods Mol Cell Biol, 5, 255

Hur E, 2000, Crystallization and aldo‐keto reductase activity of Gcy1p from Saccharomyces cerevisiae, Acta Crystallogr, 56, 763

10.1128/MCB.20.21.8157-8167.2000

10.1016/S0006-2952(97)84253-0

10.1016/S0009-2797(00)00295-7

10.1007/s002530051118

10.1016/S0167-4838(98)00217-9

10.1128/AEM.61.4.1580-1585.1995

10.1111/j.1462-5822.2007.00901.x

10.1042/bj3610163

10.1101/gad.10.5.592

10.1016/B978-0-12-380060-2.50011-8

10.1038/387s007

10.1271/bbb.61.375

10.1074/jbc.272.9.5544

Pawlowski J, 1994, Overexpression and mutagenesis of the cDNA for rat liver 3α‐hydroxysteroid/dihydrodiol dehydrogenase. Role of cysteines and tyrosines in catalysis, J Biol Chem, 269, 13502, 10.1016/S0021-9258(17)36860-6

10.1016/0167-4838(82)90402-2

10.1016/S0009-2797(00)00258-1

10.1099/00221287-136-6-1043

Van Iersel M, 1997, Purification and characterization of a novel NADP‐dependent branched‐chain alcohol dehydrogenase from Saccharomyces cerevisiae, Appl Environ Microbiol, 63, 4079, 10.1128/aem.63.10.4079-4082.1997

Vander Jagt DL, 1990, Aldehyde and aldose reductases from human placenta: heterogeneous expression of multiple enzyme forms, J Biol Chem, 265, 10 912, 10.1016/S0021-9258(19)38533-3

10.1042/bj2260669

10.1002/(SICI)1097-0061(19970915)13:11<1065::AID-YEA159>3.0.CO;2-K

10.1002/yea.320101310

10.1016/S0065-2164(08)70014-5

Wermuth B, 1985, Enzymology of Carbonyl Metabolism, 209

10.1074/jbc.M008209200

Thông tin
Thông tin xuất bản

Yeast

Tập 19 Số 12

1087-1096

Thông tin tác giả