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 language | English |
|---|---|
| Journal | Bioorganic & Medicinal Chemistry |
| Volume | 24 |
| Issue number | 1 |
| Pages (from-to) | 173-176 |
| ISSN | 0968-0896 |
| DOIs | |
| Publication status | Published - 2014 |
| Externally published | Yes |
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