Purification and cDNA Cloning of NADPH-Dependent Aldoketoreductase, Involved in Asymmetric Reduction of Methyl 4-Bromo-3-Oxobutyrate, from
            <i>Penicillium citrinum</i>
            IFO4631

Asako, Hiroyuki; Wakita, Ryuhei; Matsumura, Kenji; Shimizu, Masatoshi; Sakai, Jun

doi:10.1128/aem.71.2.1101-1104.2005

Cited by 14 publications

(5 citation statements)

References 19 publications

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“…The gld1 gene showed highest homology to an aldoketoreductase from Penicillium citrinum [14] in a blast search, not considering hypothetical proteins. The kinetic properties of GLD1 were also distinctly different from those of GLD2.…”

Section: Gld2mentioning

confidence: 99%

“…We have made a clustalw alignment of GLD1 and GLD2 of H. jecorina together with some homologous proteins for which the protein sequences have been published and some of the kinetic properties have been described. GLD1 showed highest homology to the P. citrinum KER [14], the S. cerevisiae YPR1 [15] and the S. cerevisiae GCY1 [16]. From their kinetic properties, all these proteins can be categorized as EC 1.1.1.72.…”

Section: Gld2mentioning

confidence: 99%

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Enzymes for the NADPH‐dependent reduction of dihydroxyacetone and D‐glyceraldehyde and L‐glyceraldehyde in the mould Hypocrea jecorina

et al. 2006

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Dihydroxyacetone (DHA), d-glyceraldehyde and l-glyceraldehyde can be reduced using NADPH as a cofactor to form glycerol and NADP. Enzymes catalysing this reaction are generally called NADP:glycerol dehydrogenases. NADP:glycerol dehydrogenase activity is common in moulds and filamentous fungi. Enzymes from different species of filamentous fungi have been purified and characterized. The enzymes purified from Aspergillus niger [1] and Aspergillus nidulans [2] catalyse the reversible reaction from glycerol and NADP to DHA and NADPH. For the A. niger enzyme, an equilibrium constant of 3.1-4.6 · 10 )12 m was estimated for the reaction: The mould Hypocrea jecorina (Trichoderma reesei) has two genes coding for enzymes with high similarity to the NADP-dependent glycerol dehydrogenase. These genes, called gld1 and gld2, were cloned and expressed in a heterologous host. The encoded proteins were purified and their kinetic properties characterized. GLD1 catalyses the conversion of d-glyceraldehyde and l-glyceraldehyde to glycerol, whereas GLD2 catalyses the conversion of dihydroxyacetone to glycerol. Both enzymes are specific for NADPH as a cofactor. The properties of GLD2 are similar to those of the previously described NADP-dependent glycerol-2-dehydrogenases (EC 1.1.1.156) purified from different mould species. It is a reversible enzyme active with dihydroxyacetone or glycerol as substrates. GLD1 resembles EC 1.1.1.72. It is also specific for NADPH as a cofactor but has otherwise completely different properties. GLD1 reduces d-glyceraldehyde and l-glyceraldehyde with similar affinities for the two substrates and similar maximal rates. The activity in the oxidizing reaction with glycerol as substrate was under our detection limit. Although the role of GLD2 is to facilitate glycerol formation under osmotic stress conditions, we hypothesize that GLD1 is active in pathways for sugar acid catabolism such as d-galacturonate catabolism.Abbreviations DHA, dihydroxyacetone; DHAP, dihydroxyacetone phosphate.

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Section: Gld2mentioning

confidence: 99%

Section: Gld2mentioning

confidence: 99%

Enzymes for the NADPH‐dependent reduction of dihydroxyacetone and D‐glyceraldehyde and L‐glyceraldehyde in the mould Hypocrea jecorina

et al. 2006

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“…So, the optimal reaction time for the kinetic resolution of (±)-ethyl 3-hydroxybutyrate was characterized to be 1.5 h. Chiral β-hydroxyesters are one important class of chiral drug intermediates. The synthesis of chiral β-hydroxyesters is of great importance in pharmaceutical industry [28][29][30]. Chiral EHB contains one secondary alcohol functional group and one carboxylic ester functional group.…”

Section: Effect Of Reaction Time On the Kinetic Resolution Of (±)-Ethmentioning

confidence: 99%

Functional Characterization of a Robust Marine Microbial Esterase and Its Utilization in the Stereo-Selective Preparation of Ethyl (S)-3-Hydroxybutyrate

Wang

Zhang

2016

Appl Biochem Biotechnol

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One novel microbial esterase PHE21 was cloned from the genome of Pseudomonas oryzihabitans HUP022 identified from the deep sea of the Western Pacific. PHE21 was heterologously expressed and functionally characterized to be a robust esterase which behaved high resistance to various metal ions, organic solvents, surfactants, and NaCl. Despite the fact that the two enantiomers of ethyl 3-hydroxybutyrate were hard to be enzymatically resolved before, we successfully resolved racemic ethyl 3-hydroxybutyrate through direct hydrolysis reactions and generated chiral ethyl (S)-3-hydroxybutyrate using esterase PHE21. After process optimization, the enantiomeric excess, the conversion rate, and the yield of desired product ethyl (S)-3-hydroxybutyrate could reach 99, 65, and 87 %, respectively. PHE21 is a novel marine microbial esterase with great potential in asymmetric synthesis as well as in other industries.

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“…Besides, the AKRs from Penicillium citrinum IFO4631, C . albicans , and Kluyveromyces lactis XP1461, was also used for synthesis chiral alcohols [ 22 – 24 ]. Nevertheless, the living example of the synthesis of chiral alcohols by AKRs are few compared to alcohol dehydrogenase and carbonyl reductase, and, it was deserved to be studied [ 25 , 26 ].…”

Section: Introductionmentioning

confidence: 99%

Gene mining, codon optimization and analysis of binding mechanism of an aldo-keto reductase with high activity, better substrate specificity and excellent solvent tolerance

Jiang

2021

PLoS ONE

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The biosynthesis of chiral alcohols has important value and high attention. Aldo–keto reductases (AKRs) mediated reduction of prochiral carbonyl compounds is an interesting way of synthesizing single enantiomers of chiral alcohols due to the high enantio-, chemo- and regioselectivity of the enzymes. However, relatively little research has been done on characterization and apply of AKRs to asymmetric synthesis of chiral alcohols. In this study, the AKR from Candida tropicalis MYA-3404 (C. tropicalis MYA-3404), was mined and characterized. The AKR shown wider optimum temperature and pH. The AKR exhibited varying degrees of catalytic activity for different substrates, suggesting that the AKR can catalyze a variety of substrates. It is worth mentioning that the AKR could catalytic reduction of keto compounds with benzene rings, such as cetophenone and phenoxyacetone. The AKR exhibited activity on N,N-dimethyl-3-keto-3-(2-thienyl)-1-propanamine (DKTP), a key intermediate for biosynthesis of the antidepressant drug duloxetine. Besides, the AKR still has high activity whether in a reaction system containing 10%-30% V/V organic solvent. What’s more, the AKR showed the strongest stability in six common organic solvents, DMSO, acetonitrile, ethyl acetate, isopropanol, ethanol, and methanol. And, it retains more that 70% enzyme activity after 6 hours, suggesting that the AKR has strong solvent tolerance. Furthermore, the protein sequences of the AKR and its homology were compared, and a 3D model of the AKR docking with coenzyme NADPH were constructed. And the important catalytic and binding sites were identified to explore the binding mechanism of the enzyme and its coenzyme. These properties, predominant organic solvents resistance and extensive substrate spectrum, of the AKR making it has potential applications in the pharmaceutical field.

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Purification and cDNA Cloning of NADPH-Dependent Aldoketoreductase, Involved in Asymmetric Reduction of Methyl 4-Bromo-3-Oxobutyrate, from Penicillium citrinum IFO4631

Cited by 14 publications

References 19 publications

Enzymes for the NADPH‐dependent reduction of dihydroxyacetone and D‐glyceraldehyde and L‐glyceraldehyde in the mould Hypocrea jecorina

Enzymes for the NADPH‐dependent reduction of dihydroxyacetone and D‐glyceraldehyde and L‐glyceraldehyde in the mould Hypocrea jecorina

Functional Characterization of a Robust Marine Microbial Esterase and Its Utilization in the Stereo-Selective Preparation of Ethyl (S)-3-Hydroxybutyrate

Gene mining, codon optimization and analysis of binding mechanism of an aldo-keto reductase with high activity, better substrate specificity and excellent solvent tolerance

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