Amino Acid Sequence and Characterization of Aldo-keto Reductase from Bakers’ Yeast

Nakamura, Katsumi; Kondo, Shinichi; Kawai, Yasushi; Nakajima, Nobuyoshi; Ohno, Atsuyoshi

doi:10.1271/bbb.61.375

Cited by 35 publications

(21 citation statements)

References 3 publications

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“…In addition to the decreased acetoin accumulation, the most obvious effect of BDH1 or mutated BDH1 overexpression was decreased diacetyl production. Five enzymes having in vitro diacetyl reductase activity in S. cerevisiae, including Bdh1, which reduces diacetyl as the second best substrate after acetoin, have been identified previously (9,12,24,38,43). We have shown that the overexpression or deletion of BDH1 results in a twofold decrease or increase in diacetyl levels, respectively, suggesting that Bdh1 is a rate-limiting enzyme in diacetyl reduction.…”

Section: Discussionmentioning

confidence: 71%

Engineering of 2,3-Butanediol Dehydrogenase To Reduce Acetoin Formation by Glycerol-Overproducing, Low-AlcoholSaccharomyces cerevisiae

Ehsani

Fernández

Biosca

et al. 2009

Appl Environ Microbiol

111

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Engineered Saccharomyces cerevisiae strains overexpressing GPD1, which codes for glycerol-3-phosphate dehydrogenase, and lacking the acetaldehyde dehydrogenase Ald6 display large-scale diversion of the carbon flux from ethanol toward glycerol without accumulating acetate. Although GPD1 ald6 strains have great potential for reducing the ethanol contents in wines, one major side effect is the accumulation of acetoin, having a negative sensory impact on wine. Acetoin is reduced to 2,3-butanediol by the NADH-dependent 2,3-butanediol dehydrogenase Bdh1. In order to investigate the influence of potential factors limiting this reaction, we overexpressed BDH1, coding for native NADH-dependent Bdh1, and the engineered gene BDH1 221,222,223 , coding for an NADPH-dependent Bdh1 enzyme with the amino acid changes 221 EIA 223 to 221 SRS 223, in a glycerol-overproducing wine yeast. We have shown that both the amount of Bdh1 and the NADH availability limit the 2,3-butanediol dehydrogenase reaction. During wine fermentation, however, the major limiting factor was the level of synthesis of Bdh1. Consistent with this finding, the overproduction of native or engineered Bdh1 made it possible to redirect 85 to 90% of the accumulated acetoin into 2,3-butanediol, a compound with neutral sensory characteristics. In addition, the production of diacetyl, a compound causing off-flavor in alcoholic beverages, whose production is increased in glycerol-overproducing yeast cells, was decreased by half. The production of higher alcohols and esters, which was slightly decreased or unchanged in GPD1 ald6 cells compared to that in the control cells, was not further modified in BDH1 cells. Overall, rerouting carbons toward glycerol and 2,3-butanediol represents a new milestone in the engineering of a low-alcohol yeast with desirable organoleptic features, permitting the decrease of the ethanol contents in wines by up to 3°.

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Section: Discussionmentioning

confidence: 71%

Engineering of 2,3-Butanediol Dehydrogenase To Reduce Acetoin Formation by Glycerol-Overproducing, Low-AlcoholSaccharomyces cerevisiae

Ehsani

Fernández

Biosca

et al. 2009

Appl Environ Microbiol

111

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“…Methylglyoxal pathway is found in T. thermosaccrolyticum [23], C. sphenoides [20] and S. cerevisiae [35,36]. These organisms are reported to ferment common sugars such as glucose, fructose, mannose, galactose, xylose, arabinose, lactose and cellobiose [21,23].…”

Section: Biochemical Pathways For the Production Of 12-propanediolmentioning

confidence: 99%

Microbial production and applications of 1,2-propanediol

et al. 2010

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“…PL reductase showed less than 10% identity with these yeast AKRs and differs substantially from them in substrate specificity. For example, PL reductase did not reduce aldoses such as xylose, and in contrast to AKRs from S. salmonicolor and S. cerevisiae, which show high activity toward 4-nitrobenzaldehyde (17,19,20), PL reductase showed the highest reactivity toward 2-nitrobenzaldehyde. Thus, the previously studied yeast AKRs may show little if any activity toward PL although their activity toward PL has not been reported.…”

Section: Discussionmentioning

confidence: 91%

“…The complete amino acid sequences of three kinds of yeast AKRs have been reported so far: xylose reductases (aldose reductases) from xylose-assimilating yeasts (15,16), aldehyde reductase from Sporobolomyces salmonicolor (17), and an enzyme from S. cerevisiae which shows high reactivity toward ␣-and ␤-keto esters (18,19). An amino-terminal sequence of AKR from S. cerevisiae has been reported (20).…”

Section: Discussionmentioning

confidence: 99%

Purification, Molecular Cloning, and Catalytic Activity ofSchizosaccharomyces pombe Pyridoxal Reductase

Nakano¹,

Morita²,

Yamamoto³

et al. 1999

Journal of Biological Chemistry

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Pyridoxal reductase (PL reductase), which catalyzes reduction of PL by NADPH to form pyridoxine and NADP ؉ , was purified from Schizosaccharomyces pombe. The purified enzyme was very unstable but was stabilized by low concentrations of various detergents such as Tween 40. The enzyme was a monomeric protein with the native molecular weight of 41,000 ؎ 1,600. The enzyme showed a single absorption peak at 280 nm (E 1% ‫؍‬ 10.0). PL and 2-nitrobenzaldehyde were excellent substrates, and no measurable activity was observed with short chain aliphatic aldehydes; substrate specificity of PL reductase was obviously different from those of yeast aldo-keto reductases (AKRs) so far purified. The peptide sequences of PL reductase were identical with those in a hypothetical 333-amino acid protein from S. pombe (the DDBJ/EMBL/GenBank TM accession number D89205). The gene corresponding to this protein was expressed in Escherichia coli, and the purified protein was found to have PL reductase activity. The recombinant PL reductase showed the same properties as those of native PL reductase. PL reductase showed only low sequence identities with members of AKR superfamily established to date; it shows the highest identity (18.5%) with human Shaker-related voltage-gated K ؉ channel ␤2 subunit. The elements of secondary structure of PL reductase, however, distributed similarly to those demonstrated in the three-dimensional structure of human aldose reductase except that loop A region is lost, and loop B region is extended. Amino acid residues involved in substrate binding or catalysis are also conserved. Conservation of these features, together with the major modifications, establish PL reductase as the first member of a new AKR family, AKR8.Pyridoxal reductase (PL 1 reductase) (formerly designated as PN dehydrogenase) catalyzes reduction of PL with NADPH and oxidation of PN with NADP ϩ as the reverse reaction. The enzyme was for the first time found in a budding yeast, Saccharomyces cerevisiae, i.e. bakers' (1) and brewers' (2) yeasts. Guirard and Snell (3) have purified it to homogeneity from bakers' yeast and showed that the enzyme is a monomeric protein with the molecular weight of about 33,000. They designated the enzyme as PL reductase because of the equilibrium of the enzyme reaction lying so far to formation of PN, and the substrate specificity, molecular weight, and the monomeric structure of the enzyme. The enzyme resembled chlordecone reductase (4) in optimal pH, molecular weight, specific requirement of for NADPH, and behavior toward sulfhydryl reagents, barbital, and common ketone substrates.The chlordecone reductase from human liver belongs to family 1 of the AKR superfamily.2 The AKRs form an expanding oxidoreductase superfamily classified into seven families containing a variety of monomeric oxidoreductases such as aldehyde and aldose reductases, hydroxysteroid dehydrogenases, chalcone reductases, Shaker-related voltage-gated K ϩ channel ␤2 subunits, and aflatoxin B 1 -aldehyde reductases (5). Because the nomenclature ...

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Amino Acid Sequence and Characterization of Aldo-keto Reductase from Bakers’ Yeast

Cited by 35 publications

References 3 publications

Engineering of 2,3-Butanediol Dehydrogenase To Reduce Acetoin Formation by Glycerol-Overproducing, Low-AlcoholSaccharomyces cerevisiae

Engineering of 2,3-Butanediol Dehydrogenase To Reduce Acetoin Formation by Glycerol-Overproducing, Low-AlcoholSaccharomyces cerevisiae

Microbial production and applications of 1,2-propanediol

Purification, Molecular Cloning, and Catalytic Activity ofSchizosaccharomyces pombe Pyridoxal Reductase

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