Essential role of the small subunit of thermostable glucose dehydrogenase from Burkholderia cepacia

• Published in: Biotechnology letters Vol. 26; no. 22; pp. 1757 - 1761
• Main Authors: YAMAOKA, Hideaki, FERRI, Stefano, FUJIKAWA, Masako, SODE, Koji
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer 01.11.2004; Springer Nature B.V
• Subjects: Amino Acid Sequence; Amino acids; Biological and medical sciences; Biotechnology; Burkholderia cepacia; Burkholderia cepacia - enzymology; Burkholderia cepacia - genetics; Dehydrogenase; E coli; Enzyme Activation; Enzyme Stability; Escherichia coli - enzymology; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Glucose 1-Dehydrogenase - analysis; Glucose 1-Dehydrogenase - biosynthesis; Glucose 1-Dehydrogenase - chemistry; Molecular Sequence Data; Molecular Weight; Protein Subunits - chemistry; Protein Subunits - metabolism; Proteins; Recombinant Proteins - chemistry; Recombinant Proteins - metabolism; Structure-Activity Relationship; Temperature; Translocation; glucose dehydrogenase; Arginine; Enzyme; Burkholderia cepacia; Chaperone; twin-arginine translocase; Bacteria; Glucose 1-dehydrogenase; Oxidoreductases; Subunit; flavoenzyme; Thermal stability
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-004-4582-0

The co-expression in Escherichia coli of the gamma-subunit and the catalytic alpha-subunit of the thermostable glucose dehydrogenase (GDH) from Burkholderia cepacia sp. SM4 produced 12.7 U GDH activity mg(-1) protein. A 47-amino acid, twin-arginine translocase signal peptide was identified at the amino terminus of the gamma-subunit. The expression of the alpha-subunit in the absence of the gamma-subunit or the gamma-subunit signal peptide failed to produce any detectable GDH protein or activity. The gamma-subunit may be a chaperone-like component that assists folding of the alpha-subunit polypeptide to the active form and its translocation to the periplasm.