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Chengcheng Yang - One of the best experts on this subject based on the ideXlab platform.

Yan Liu - One of the best experts on this subject based on the ideXlab platform.

  • Stereoselective Bioreduction of Ethyl 3-Oxo-3-(2-Thienyl) Propanoate Using the Short-Chain Dehydrogenase/Reductase ChKRED12.
    Journal of microbiology and biotechnology, 2019
    Co-Authors: Zhi-qiang Ren, Yan Liu, Xiao-qiong Pei
    Abstract:

    Ethyl (S)-3-hydroxy-3-(2-thienyl)propanoate((S)-HEES)acts as a key chiral intermediate for the blockbuster antidepressant drug duloxetine, which canbe achieved viathe stereoselective bioreduction ofethyl 3-oxo-3-(2-thienyl) propanoate (KEES) that containsa 3-oxoacyl structure.The sequences of the Short-Chain Dehydrogenase/reductases from Chryseobacterium sp. CA49 were analyzed, and the putative3-oxoacyl-acyl-carrier-protein reductase, ChKRED12, was able to stereoselectivelycatalyze theNADPH-dependent reduction to produce (S)-HEES.The reductase activity of ChKRED12 towardsothersubstrates with 3-oxoacyl structure were confirmed with excellent stereoselectivity (>99% enantiomeric excess) in most cases. When coupled with a cofactor recycling system using glucose Dehydrogenase, the ChKRED12 was able to catalyze the complete conversion of 100 g/l KEES within 12h, yielding the enantiopure product with >99% ee, showing a remarkable potential to produce (S)-HEES.

  • stereoselective bioreduction of ethyl 3 oxo 3 2 thienyl propanoate using the short chain Dehydrogenase reductase chkred12
    Journal of Microbiology and Biotechnology, 2019
    Co-Authors: Zhi-qiang Ren, Yan Liu, Xiao-qiong Pei
    Abstract:

    Ethyl (S)-3-hydroxy-3-(2-thienyl)propanoate((S)-HEES)acts as a key chiral intermediate for the blockbuster antidepressant drug duloxetine, which canbe achieved viathe stereoselective bioreduction ofethyl 3-oxo-3-(2-thienyl) propanoate (KEES) that containsa 3-oxoacyl structure.The sequences of the Short-Chain Dehydrogenase/reductases from Chryseobacterium sp. CA49 were analyzed, and the putative3-oxoacyl-acyl-carrier-protein reductase, ChKRED12, was able to stereoselectivelycatalyze theNADPH-dependent reduction to produce (S)-HEES.The reductase activity of ChKRED12 towardsothersubstrates with 3-oxoacyl structure were confirmed with excellent stereoselectivity (>99% enantiomeric excess) in most cases. When coupled with a cofactor recycling system using glucose Dehydrogenase, the ChKRED12 was able to catalyze the complete conversion of 100 g/l KEES within 12h, yielding the enantiopure product with >99% ee, showing a remarkable potential to produce (S)-HEES.

  • characterization of a robust anti prelog short chain Dehydrogenase reductase chkred20 from chryseobacterium sp ca49
    Journal of Molecular Catalysis B-enzymatic, 2014
    Co-Authors: Tuoxian Tang, Yan Liu
    Abstract:

    ChKRED20 is a Short-Chain Dehydrogenase/reductase (SDR) cloned from Chryseobacterium sp. CA49 for the anti-Prelog bioreduction of 3,5-bis(trifluoromethyl)acetophenone to produce the chiral alcohol intermediate for aprepitant. Purified ChKRED20 showed broad pH adaptability and stability with 91% of the maximal activity retained at pH 10.0. The temperature dependence of activity reached the maxima at 50 degrees C. Its half-lives of thermal inactivation were 163 and 9.8 h at 40 degrees C and 50 degrees C, respectively. The enzyme was resistant to a variety of metal ions, additives, and organic solvents. The enzymatic activity could be enhanced by the addition of particular metal ions or detergents to up to 168%. ChKRED20 also displayed good activity and excellent an ti-Prelog stereoselectivity toward a spectrum of acetophenone derivatives, providing chiral alcohols with >99% ee for the majority of the substrates. (C) 2014 Elsevier B.V. All rights reserved.

  • Characterization of a robust anti-Prelog Short-Chain Dehydrogenase/reductase ChKRED20 from Chryseobacterium sp. CA49
    Journal of Molecular Catalysis B: Enzymatic, 2014
    Co-Authors: Tang Tuoxian, Yan Liu
    Abstract:

    ChKRED20 is a Short-Chain Dehydrogenase/reductase (SDR) cloned from Chryseobacterium sp. CA49 for the anti-Prelog bioreduction of 3,5-bis(trifluoromethyl)acetophenone to produce the chiral alcohol intermediate for aprepitant. Purified ChKRED20 showed broad pH adaptability and stability with 91% of the maximal activity retained at pH 10.0. The temperature dependence of activity reached the maxima at 50 degrees C. Its half-lives of thermal inactivation were 163 and 9.8 h at 40 degrees C and 50 degrees C, respectively. The enzyme was resistant to a variety of metal ions, additives, and organic solvents. The enzymatic activity could be enhanced by the addition of particular metal ions or detergents to up to 168%. ChKRED20 also displayed good activity and excellent an ti-Prelog stereoselectivity toward a spectrum of acetophenone derivatives, providing chiral alcohols with >99% ee for the majority of the substrates. (C) 2014 Elsevier B.V. All rights reserved.

Xiaohong Yang - One of the best experts on this subject based on the ideXlab platform.

Natalia Y Kedishvili - One of the best experts on this subject based on the ideXlab platform.

  • Medium- and Short-Chain Dehydrogenase/reductase gene and protein families : Medium-chain and Short-Chain Dehydrogenases/reductases in retinoid metabolism.
    Cellular and molecular life sciences : CMLS, 2008
    Co-Authors: Xavier Parés, Natalia Y Kedishvili, Jaume Farrés, Gregg Duester
    Abstract:

    Retinoic acid (RA), the most active retinoid, is synthesized in two steps from retinol. The first step, oxidation of retinol to retinaldehyde, is catalyzed by cytosolic alcohol Dehydrogenases (ADHs) of the medium-chain Dehydrogenase/reductase (MDR) superfamily and microsomal retinol Dehydrogenases (RDHs) of the Short-Chain Dehydrogenase/reductase (SDR) superfamily. The second step, oxidation of retinaldehyde to RA, is catalyzed by several aldehyde Dehydrogenases. ADH1 and ADH2 are the major MDR enzymes in liver retinol detoxification, while ADH3 (less active) and ADH4 (most active) participate in RA generation in tissues. Several NAD+- and NADP+-dependent SDRs are retinoid active. Their in vivo contribution has been demonstrated in the visual cycle (RDH5, RDH12), adult retinoid homeostasis (RDH1) and embryogenesis (RDH10). K m values for most retinoid-active ADHs and RDHs are close to 1 μM or lower, suggesting that they participate physiologically in retinol/retinaldehyde interconversion. Probably none of these enzymes uses retinoids bound to cellular retinol-binding protein, but only free retinoids. The large number of enzymes involved in the two directions of this step, also including aldo-keto reductases, suggests that retinaldehyde levels are strictly regulated.

  • Human retinol Dehydrogenase 13 (RDH13) is a mitochondrial Short-Chain Dehydrogenase/reductase with a retinaldehyde reductase activity
    The FEBS journal, 2007
    Co-Authors: Olga V Belyaeva, Olga V. Korkina, Anton V. Stetsenko, Natalia Y Kedishvili
    Abstract:

    Retinol Dehydrogenase 13 (RDH13) is a recently identified Short-Chain Dehydrogenase/reductase related to microsomal retinoid oxidoreductase RDH11. In this study, we examined the distribution of RDH13 in human tissues, determined its subcellular localization and characterized the substrate and cofactor specificity of purified RDH13 in order to better understand its properties. The results of this study demonstrate that RDH13 exhibits a wide tissue distribution and, by contrast with other members of the RDH11-like group of Short-Chain Dehydrogenases/reductases, is a mitochondrial rather than a microsomal protein. Protease protection assays suggest that RDH13 is localized on the outer side of the inner mitochondrial membrane. Kinetic analysis of the purified protein shows that RDH13 is catalytically active and recognizes retinoids as substrates. Similar to the microsomal RDHs, RDH11, RDH12 and RDH14, RDH13 exhibits a much lower Km value for NADPH than for NADH and has a greater catalytic efficiency in the reductive than in the oxidative direction. The localization of RDH13 at the entrance to the mitochondrial matrix suggests that it may function to protect mitochondria against oxidative stress associated with the highly reactive retinaldehyde produced from dietary β-carotene.

  • properties of short chain Dehydrogenase reductase ralr1 characterization of purified enzyme its orientation in the microsomal membrane and distribution in human tissues and cell lines
    Biochemistry, 2003
    Co-Authors: Olga V Belyaeva, Peter S Nelson, Anton V. Stetsenko, Natalia Y Kedishvili
    Abstract:

    Recently, we reported the first biochemical characterization of a novel member of the Short-Chain Dehydrogenase/reductase superfamily, retinal reductase 1 (RalR1) (Kedishvili et al. (2002) J. Biol. Chem. 277, 28909-28915). In the present study, we purified the recombinant enzyme from the microsomal membranes of insect Sf9 cells, determined its catalytic efficiency for the reduction of retinal and the oxidation of retinol, established its transmembrane topology, and examined the distribution of RalR1 in human tissues and cell lines. Purified RalR1-His(6) exhibited the apparent K(m) values for all-trans-retinal and all-trans-retinol of 0.12 and 0.6 microM, respectively. The catalytic efficiency (k(cat)/K(m)) for the reduction of all-trans-retinal (150,000 min(-1) mM(-1)) was 8-fold higher than that for the oxidation of all-trans-retinol (18,000 min(-1) mM(-1)). Protease protection assays and site-directed mutagenesis suggested that the enzyme is anchored in the membrane by the N-terminal signal-anchor domain, with the majority of the polypeptide chain located on the cytosolic side of the membrane. An important feature that prevented the translocation of RalR1 across the membrane was the positively charged R(25)K motif flanking the N-terminal signal-anchor. The cytosolic orientation of RalR1 suggested that, in intact cells, the enzyme would function predominantly as a reductase. Western blot analysis revealed that RalR1 is expressed in a wide variety of normal human tissues and cancer cell lines. The expression pattern and the high catalytic efficiency of RalR1 are consistent with the hypothesis that RalR1 contributes to the reduction of retinal in various human tissues.

  • evidence that the human gene for prostate short chain Dehydrogenase reductase psdr1 encodes a novel retinal reductase ralr1
    Journal of Biological Chemistry, 2002
    Co-Authors: Natalia Y Kedishvili, Olga V Chumakova, Sergei V Chetyrkin, Olga V Belyaeva, Elena A Lapshina, Daniel W Lin, Masazumi Matsumura, Peter S Nelson
    Abstract:

    All-trans-retinoic acid is a metabolite of vitamin A (all-trans-retinol) that functions as an activating ligand for a family of nuclear retinoic acid receptors. The intracellular levels of retinoic acid in tissues are tightly regulated, although the mechanisms underlying the control of retinoid metabolism at the level of specific enzymes are not completely understood. In this report we present the first characterization of the retinoid substrate specificity of a novel Short-Chain Dehydrogenase/reductase (SDR) encoded by RalR1/PSDR1, a cDNA recently isolated from the human prostate (Lin, B., White, J. T., Ferguson, C., Wang, S., Vessella, R., Bumgarner, R., True, L. D., Hood, L., and Nelson, P. S. (2001) Cancer Res. 61, 1611-1618). We demonstrate that RalR1 exhibits an oxidoreductive catalytic activity toward retinoids, but not steroids, with at least an 800-fold lower apparent K(m) values for NADP+ and NADPH versus NAD+ and NADH as cofactors. The enzyme is approximately 50-fold more efficient for the reduction of all-trans-retinal than for the oxidation of all-trans-retinol. Importantly, RalR1 reduces all-trans-retinal in the presence of a 10-fold molar excess of cellular retinol-binding protein type I, which is believed to sequester all-trans-retinal from nonspecific enzymes. As shown by immunostaining of human prostate and LNCaP cells with monoclonal anti-RalR1 antibodies, the enzyme is highly expressed in the epithelial cell layer of human prostate and localizes to the endoplasmic reticulum. The enzymatic properties and expression pattern of RalR1 in prostate epithelium suggest that it might play a role in the regulation of retinoid homeostasis in human prostate.

  • Cloning of the human RoDH-related short chain Dehydrogenase gene and analysis of its structure.
    Chemico-Biological Interactions, 2001
    Co-Authors: Natalia Y Kedishvili, Olga V Belyaeva, Wendy H. Gough
    Abstract:

    We have previously characterized the first human NAD(+)-dependent short chain Dehydrogenase capable of oxidizing all-trans-retinol and androgens, and found only in the liver and skin. In a search for related human enzymes, we identified a partial open reading frame, which exhibited >60% sequence identity to human RoDH-4. The full-length cDNA for this enzyme was determined in our laboratory by 5'-RACE PCR and was found to be identical to the recently reported novel type of oxidative human 3alpha-hydroxysteroid Dehydrogenase (3alpha-HSD). Analysis of the genomic structure revealed that the gene for RoDH-like 3alpha-HSD has four translated exons and, possibly, a fifth exon that codes for the 5'-untranslated region. The gene for RoDH-4 appears to have only four exons. The positions of exon-intron boundaries and the sizes of the protein coding regions are identical in 3alpha-HSD and RoDH-4. Moreover, both genes are mapped to chromosome 12q13, and are located in a close proximity to each other. Both genes appear to have satellite pseudogenes. Thus, RoDH-4 and 3alpha-HSD genes share similar structural organization and cluster on human chromosome 12, near the gene for 11-cis retinol Dehydrogenase.

Peter S Nelson - One of the best experts on this subject based on the ideXlab platform.

  • properties of short chain Dehydrogenase reductase ralr1 characterization of purified enzyme its orientation in the microsomal membrane and distribution in human tissues and cell lines
    Biochemistry, 2003
    Co-Authors: Olga V Belyaeva, Peter S Nelson, Anton V. Stetsenko, Natalia Y Kedishvili
    Abstract:

    Recently, we reported the first biochemical characterization of a novel member of the Short-Chain Dehydrogenase/reductase superfamily, retinal reductase 1 (RalR1) (Kedishvili et al. (2002) J. Biol. Chem. 277, 28909-28915). In the present study, we purified the recombinant enzyme from the microsomal membranes of insect Sf9 cells, determined its catalytic efficiency for the reduction of retinal and the oxidation of retinol, established its transmembrane topology, and examined the distribution of RalR1 in human tissues and cell lines. Purified RalR1-His(6) exhibited the apparent K(m) values for all-trans-retinal and all-trans-retinol of 0.12 and 0.6 microM, respectively. The catalytic efficiency (k(cat)/K(m)) for the reduction of all-trans-retinal (150,000 min(-1) mM(-1)) was 8-fold higher than that for the oxidation of all-trans-retinol (18,000 min(-1) mM(-1)). Protease protection assays and site-directed mutagenesis suggested that the enzyme is anchored in the membrane by the N-terminal signal-anchor domain, with the majority of the polypeptide chain located on the cytosolic side of the membrane. An important feature that prevented the translocation of RalR1 across the membrane was the positively charged R(25)K motif flanking the N-terminal signal-anchor. The cytosolic orientation of RalR1 suggested that, in intact cells, the enzyme would function predominantly as a reductase. Western blot analysis revealed that RalR1 is expressed in a wide variety of normal human tissues and cancer cell lines. The expression pattern and the high catalytic efficiency of RalR1 are consistent with the hypothesis that RalR1 contributes to the reduction of retinal in various human tissues.

  • evidence that the human gene for prostate short chain Dehydrogenase reductase psdr1 encodes a novel retinal reductase ralr1
    Journal of Biological Chemistry, 2002
    Co-Authors: Natalia Y Kedishvili, Olga V Chumakova, Sergei V Chetyrkin, Olga V Belyaeva, Elena A Lapshina, Daniel W Lin, Masazumi Matsumura, Peter S Nelson
    Abstract:

    All-trans-retinoic acid is a metabolite of vitamin A (all-trans-retinol) that functions as an activating ligand for a family of nuclear retinoic acid receptors. The intracellular levels of retinoic acid in tissues are tightly regulated, although the mechanisms underlying the control of retinoid metabolism at the level of specific enzymes are not completely understood. In this report we present the first characterization of the retinoid substrate specificity of a novel Short-Chain Dehydrogenase/reductase (SDR) encoded by RalR1/PSDR1, a cDNA recently isolated from the human prostate (Lin, B., White, J. T., Ferguson, C., Wang, S., Vessella, R., Bumgarner, R., True, L. D., Hood, L., and Nelson, P. S. (2001) Cancer Res. 61, 1611-1618). We demonstrate that RalR1 exhibits an oxidoreductive catalytic activity toward retinoids, but not steroids, with at least an 800-fold lower apparent K(m) values for NADP+ and NADPH versus NAD+ and NADH as cofactors. The enzyme is approximately 50-fold more efficient for the reduction of all-trans-retinal than for the oxidation of all-trans-retinol. Importantly, RalR1 reduces all-trans-retinal in the presence of a 10-fold molar excess of cellular retinol-binding protein type I, which is believed to sequester all-trans-retinal from nonspecific enzymes. As shown by immunostaining of human prostate and LNCaP cells with monoclonal anti-RalR1 antibodies, the enzyme is highly expressed in the epithelial cell layer of human prostate and localizes to the endoplasmic reticulum. The enzymatic properties and expression pattern of RalR1 in prostate epithelium suggest that it might play a role in the regulation of retinoid homeostasis in human prostate.

  • Isolation and characterization of the murine prostate Short-Chain Dehydrogenase/reductase 1 (Psdr1) gene, a new member of the Short-Chain steroid Dehydrogenase/reductase family.
    Gene, 2002
    Co-Authors: Stacy Moore, Colin Pritchard, Biaoyang Lin, Camari Ferguson, Peter S Nelson
    Abstract:

    We report the isolation and characterization of a complementary DNA (cDNA) encoding a novel member of the Short-Chain Dehydrogenase/reductase (SDR) gene family that we have designated murine prostate Short-Chain Dehydrogenase/reductase 1 (Psdr1). Psdr1 was cloned as a 3.2 kbp transcript from mouse testis cDNA based on the sequence of the recently described androgen-regulated human PSDR1 gene (Cancer Res. 61 (2001) 1611). The putative protein encoded by Psdr1 consists of 316 amino acids with 85% identity to human PSDR1. A search against the BLOCKS database of conserved protein motifs indicates that Psdr1 retains features essential for SDR function. Northern analyses demonstrate that Psdr1 is highly expressed in the murine testis and liver and exhibits several isoforms. Cloning and sequence analysis of the putative Psdr1 promoter region identified motifs with homology to the consensus androgen response element and progesterone response element. The Psdr1 gene was mapped to mouse chromosome 12q31-34, which has synteny with the human PSDR1 chromosomal location (14q23-24.3). Together, these data describe a new member of the SDR gene family that may be involved in the tissue-specific metabolism of retinoids or steroid hormones.

  • Isolation and characterization of the murine prostate Short-Chain Dehydrogenase/reductase 1 (Psdr1) gene, a new member of the Short-Chain steroid Dehydrogenase/reductase family.
    Gene, 2002
    Co-Authors: Stacy Moore, Colin Pritchard, Biaoyang Lin, Camari Ferguson, Peter S Nelson
    Abstract:

    We report the isolation and characterization of a complementary DNA (cDNA) encoding a novel member of the Short-Chain Dehydrogenase/reductase (SDR) gene family that we have designated murine prostate Short-Chain Dehydrogenase/reductase 1 (Psdr1). Psdr1 was cloned as a 3.2 kbp transcript from mouse testis cDNA based on the sequence of the recently described androgen-regulated human PSDR1 gene (Cancer Res. 61 (2001) 1611). The putative protein encoded by Psdr1 consists of 316 amino acids with 85% identity to human PSDR1. A search against the BLOCKS database of conserved protein motifs indicates that Psdr1 retains features essential for SDR function. Northern analyses demonstrate that Psdr1 is highly expressed in the murine testis and liver and exhibits several isoforms. Cloning and sequence analysis of the putative Psdr1 promoter region identified motifs with homology to the consensus androgen response element and progesterone response element. The Psdr1 gene was mapped to mouse chromosome 12q31‐34, which has synteny with the human PSDR1 chromosomal location (14q23‐24.3). Together, these data describe a new member of the SDR gene family that may be involved in the tissue-specific metabolism of retinoids or steroid hormones. q 2002 Published by Elsevier Science B.V.

  • Prostate Short-Chain Dehydrogenase Reductase 1 (PSDR1): A New Member of the Short-Chain Steroid Dehydrogenase/Reductase Family Highly Expressed in Normal and Neoplastic Prostate Epithelium
    Cancer research, 2001
    Co-Authors: Biaoyang Lin, Camari Ferguson, James T White, Shunyou Wang, Robert Vessella, Roger Bumgarner, Lawrence D True, Leroy Hood, Peter S Nelson
    Abstract:

    Genes regulated by androgenic hormones are of critical importance for the normal physiological function of the human prostate gland, and they contribute to the development and progression of prostate carcinoma. We used cDNA microarrays comprised of prostate-derived cDNAs to profile transcripts regulated by androgens in prostate cancer cells. This study identified a novel gene that we have designated prostate Short-Chain Dehydrogenase/reductase 1 ( PSDR1 ), that exhibits increased expression on exposure to androgens in the LNCaP prostate cancer cell line. Northern analysis demonstrated that PSDR1 is highly expressed in the prostate gland relative to other normal human tissues. The PSDR1 cDNA and putative protein exhibit homology to the family of Short-Chain Dehydrogenase/reductase enzymes and thus identify a new member of this family. Cloning and analysis of the putative PSDR1 promoter region identified a potential androgen-response element. We used a radiation-hybrid panel to map the PSDR1 gene to chromosome 14q23-24.3. In situ hybridization localizes PSDR1 expression to normal and neoplastic prostate epithelium. These results identify a new gene involved in the androgen receptor-regulated gene network of the human prostate that may play a role in the pathogenesis of prostate carcinoma.