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Title of Journal: Mol Biotechnol

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Abbravation: Molecular Biotechnology

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Springer US

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DOI

10.1007/s00705-001-0778-y

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1559-0305

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Heterologous Overexpression Purification and Char

Authors: AnnKathrin Liliensiek Jennifer Cassidy Gabriele Gucciardo Cliadhna Whitely Francesca Paradisi
Publish Date: 2013/05/05
Volume: 55, Issue: 2, Pages: 143-149
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Abstract

Replacement of chemical steps with biocatalytic ones is becoming increasingly more interesting due to the remarkable catalytic properties of enzymes such as their wide range of substrate specificities and variety of chemo stereo and regioselective reactions This study presents characterisation of an alcohol dehydrogenase ADH from the halophilic archaeum Halobacterium sp NRC1 HsADH2 A hexahistidinetagged recombinant version of HsADH2 HisHsADH2 was heterologously overexpressed in Haloferax volcanii The enzyme was purified in one step by immobilised Niaffinity chromatography HisHsADH2 was halophilic and mildly thermophilic with optimal activity for ethanol oxidation at 4 M KCl around 60 °C and pH 100 The enzyme was extremely stable retaining 80  activity after 30 days HisHsADH2 showed preference for NADPH but interestingly retained 60  activity towards NADH The enzyme displayed broad substrate specificity with maximum activity obtained for 1propanol The enzyme also accepted secondary alcohols such as 2butanol and even 1phenylethanol In the reductive reaction working conditions for HisHsADH2 were optimised for acetaldehyde and found to be 4 M KCl and pH 60 HisHsADH2 displayed intrinsic organic solvent tolerance which is highly relevant for biotechnological applicationsFP acknowledges funding under the Science Foundation Ireland Research Framework Programme and the EPA Kind donations of the strain H sp NRC1 by Prof P Engel and the expression system H volcanii H1209H1325/pTA963 by Dr T Allers are gratefully acknowledged


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Other Papers In This Journal:

  1. Production and Characterization of a Single-Chain Fv Antibody–Alkaline Phosphatase Fusion Protein Specific for Clenbuterol
  2. Improvement of Cephalosporin C Production by Recombinant DNA Integration in Acremonium chrysogenum
  3. Codon Optimization of the “Bos Taurus Chymosin” Gene for the Production of Recombinant Chymosin in Pichia pastoris
  4. Enhanced Extracellular Production of Heterologous Proteins in Bacillus subtilis by Deleting the C-terminal Region of the SecA Secretory Machinery
  5. Enhanced Production of Human Recombinant Proteins from CHO cells Grown to High Densities in Macroporous Microcarriers
  6. Molecular Cloning and Characterization of a Malic Enzyme Gene from the Oleaginous Yeast Lipomyces starkeyi
  7. Hydrophobic Substitution of Surface Residues Affects Lipase Stability in Organic Solvents
  8. Identification of Suitable Endogenous Normalizers for qRT-PCR Analysis of Plasma microRNA Expression in Essential Hypertension
  9. Biosynthesis of Resveratrol in Blastospore of the Macrofungus Tremella fuciformis
  10. Heterologous Expression and Characterization of an Alcohol Dehydrogenase from the Archeon Thermoplasma acidophilum
  11. Plasma Antioxidants and Human Aging: A Study on Healthy Elderly Tunisian Population
  12. Importance of the residue Asp 290 on chain length selectivity and catalytic efficiency of recombinant Staphylococcus simulans lipase expressed in E. coli
  13. Fahd Al-Mulla (ed). Formalin-Fixed Paraffin-Embedded Tissue. Methods and Protocols. Methods in Molecular Biology 724. The Humana Press, ISBN 978-1-61779-054-6
  14. The Peptide Derived from erbB2 Auto-Inhibitor Herstatin Shared in the Same Epitope and Function with Functional Antibody 2C4
  15. End Joining-Mediated Gene Expression in Mammalian Cells Using PCR-Amplified DNA Constructs that Contain Terminator in Front of Promoter
  16. Identification of Functional Regions in the Rhodospirillum rubrum l -Asparaginase by Site-Directed Mutagenesis
  17. Expression of an Acid Urease with Urethanase Activity in E. coli and Analysis of Urease Gene

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