Overexpression of an archaeal geranylgeranyl diphosphate synthase in Escherichia coli cells

• Published in: Bioscience, biotechnology, and biochemistry Vol. 62; no. 6; pp. 1243 - 1246
• Main Authors: Ohto, C. (Toyota Motor Corp., Aichi (Japan)), Nakane, H, Hemmi, H, Ohnuma, S, Obata, S, Nishino, T
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience, Biotechnology, and Agrochemistry 01.06.1998; Japan Society for Bioscience Biotechnology and Agrochemistry
• Subjects: Alkyl and Aryl Transferases - biosynthesis; Alkyl and Aryl Transferases - genetics; Archaea; ATP-Binding Cassette Transporters; Bacterial Proteins - isolation & purification; Biological and medical sciences; Biotechnology; Carrier Proteins - isolation & purification; Chemical Fractionation; ENZIMAS; ENZYME; ENZYMES; ESCHERICHIA COLI; Escherichia coli Proteins; Farnesyltranstransferase; Fundamental and applied biological sciences. Psychology; fusion protein; Gene Expression; Genetic engineering; Genetic technics; Maltose-Binding Proteins; Methods. Procedures. Technologies; Modification of gene expression level; Monosaccharide Transport Proteins; prenyldiphosphate; PROTEINAS RECOMBINANTES; PROTEINE RECOMBINANTE; recombinant enzyme; Recombinant Fusion Proteins - biosynthesis; RECOMBINANT PROTEINS; Sulfolobus acidocaldarius; TRANSFERASAS; TRANSFERASE; TRANSFERASES; Heterologous system; Oligomerization; Purification; Archaeobacteria; Enzyme; Transferases; Escherichia coli; Reaction product; Gene overexpression; Gene expression; Farnesyltranstransferase; Sulfolobaceae; Sulfolobus acidocaldarius; Enzymatic activity; Structure activity relation; Bacteria; Sulfolobales; Recombinant protein; Fusion protein; Enterobacteriaceae
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.62.1243

An archaeal geranylgeranyl diphosphate synthase was overexpressed in Escherichia coli cells as fusion proteins. These fusion proteins retained their thermostability and had higher specific activity than did a partially purified native enzyme Previously reported. We purified 24.3 mg of MBP (maltose-binding protein)-fusion protein and 5.4 mg of GST (glutathione S-transferase)-fusion protein from a one-liter culture of E. coli. The MBP-fusion protein existed in dimer, tetramer, octamer, or dodecamer form, and their product specificities were altered according to the oligomerization. The MBP-fusion protein has protease-sensitive sites in the portion corresponding to geranylgeranyl diphosphate synthase