pH-rate profiles of L-arabinitol 4-dehydrogenase from Hypocrea jecorina and its application in L-xylulose production

Manish Kumar Tiwari, Raushan Kumar Singh, Hui Gao, Taesu Kim, Suhwan Chang, Han S. Kim, Jung-Kul Lee

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

L-Arabinitol 4-dehydrogenase (LAD) from Hypocrea jecorina (HjLAD) was cloned and overexpressed in Escherichia coil BL21 (DE3). The kinetics of L-arabinitol oxidation by NAD(+), catalyzed by HjLAD, was studied within the pH range of 7.0-9.5 at 25 degrees C. The turnover number (k(cat)) and the catalytic efficiency (k(cat)/K-m) were 4200 min(-1) and 290 mM(-1) min(-1), respectively. HjLAD showed the highest turnover number and catalytic efficiency among all previously characterized LADs. In further application of HjLAD, rare L-sugar L-xylulose was produced by the enzymatic oxidation of arabinitol to give a yield of approximately 86%. (C) 2013 Elsevier Ltd. All rights reserved.
Original languageEnglish
JournalBioorganic & Medicinal Chemistry
Volume24
Issue number1
Pages (from-to)173-176
ISSN0968-0896
DOIs
Publication statusPublished - 2014
Externally publishedYes

Keywords

  • BIOCHEMISTRY
  • CHEMISTRY,
  • LIVER SORBITOL DEHYDROGENASE
  • YEAST ALCOHOL-DEHYDROGENASE
  • L-LYXOSE
  • CLONING
  • GENE
  • EXPRESSION
  • XYLITOL
  • XYLOSE
  • BIOCATALYSIS
  • INHIBITION
  • Rare sugar
  • L-Xylulose
  • PH-Dependent kinetics
  • Molecular dynamics
  • pH-Dependent kinetics
  • Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Endospore-forming Gram-Positives [07810] Bacillus subtilis species
  • Facultatively Anaerobic Gram-Negative Rods Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Enterobacteriaceae [06702] Escherichia coli species expression system strain-BL21
  • Fungi Plantae (Fungi, Microorganisms, Nonvascular Plants, Plants) - Ascomycetes [15100] Hypocrea jecorina species Neurospora crassa species
  • Fungi Plantae (Fungi, Microorganisms, Nonvascular Plants, Plants) - Fungi Imperfecti or Deuteromycetes [15500] Penicillium chrysogenum species Aspergillus niger species Aspergillus oryzae species Trichoderma longibrachiatum species
  • Hypocrea jecorina LAD gene [Ascomycetes] Hypocrea jecorina L-arabinitol 4-dehydrogenase gene
  • arabinitol 2152-56-9
  • L-arabinitol 4-dehydrogenase expression EC 1.1.1.12
  • L-sugar
  • L-xylulose 527-50-4 oxidation
  • NAD 53-84-9
  • 03502, Genetics - General
  • 03504, Genetics - Plant
  • 10062, Biochemistry studies - Nucleic acids, purines and pyrimidines
  • 10064, Biochemistry studies - Proteins, peptides and amino acids
  • 10802, Enzymes - General and comparative studies: coenzymes
  • 31000, Physiology and biochemistry of bacteria
  • 31500, Genetics of bacteria and viruses
  • 51518, Plant physiology - Enzymes
  • Enzymology
  • Molecular Genetics
  • catalytic efficiency
  • enzymatic oxidation
  • enzyme kinetics
  • pH-rate profile
  • substrate turnover number
  • Biochemistry and Molecular Biophysics
  • 3M6I Protein databank amino acid sequence
  • cloning laboratory techniques, genetic techniques
  • Facultatively Anaerobic Gram-Negative Rods Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Enterobacteriaceae [06702] Escherichia coli species strain-BL21
  • Fungi Plantae (Fungi, Microorganisms, Nonvascular Plants, Plants) - Ascomycetes [15100] Hypocrea jecorina species
  • Hypocrea jecorina HjLAD gene [Ascomycetes]
  • levo-arabinitol 4-dehydrogenase
  • levo-xylulose
  • 10060, Biochemistry studies - General
  • 51522, Plant physiology - Chemical constituents
  • arabinitol
  • levo arabinitol 4 dehydrogenase
  • levo xylulose
  • oxidoreductase
  • unclassified drug
  • xylulose
  • affinity chromatography
  • alcohol oxidation
  • alkalinity
  • amino acid sequence
  • article
  • crystal structure
  • dehydrogenation
  • enzyme active site
  • enzyme activity
  • Escherichia coli
  • gel filtration chromatography
  • Hypocrea jecorina
  • kinetics
  • Michaelis Menten kinetics
  • microenvironment
  • molecular weight
  • nonhuman
  • oxidation
  • pH
  • polyacrylamide gel electrophoresis
  • protein secondary structure
  • sequence alignment
  • site directed mutagenesis
  • steady state
  • Trichoderma longibrachiatum
  • turnover time
  • l-Xylulose
  • ALDOSE reductase

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