Overexpression of an archaeal geranylgeranyl diphosphate synthase in Escherichia coli cells.
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 proteins 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.
Ohto
C
C
Bio Research Laboratory, Toyota Motor Corporation, Japan. ohto@square.mk.toyota.co.jp
Nakane
H
H
Hemmi
H
H
Ohnuma
S
S
Obata
S
S
Nishino
T
T
D018528:ATP-Binding Cassette Transporters; D019883:Alkyl and Aryl Transferases / Q000096:biosynthesis / Q000235:genetics*; D001426:Bacterial Proteins / Q000302:isolation & purification; D002352:Carrier Proteins / Q000302:isolation & purification; D005591:Chemical Fractionation; D004926:Escherichia coli; D029968:Escherichia coli Proteins; D051231:Farnesyltranstransferase; D015870:Gene Expression; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D016945:Sulfolobus acidocaldarius
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D011993:Recombinant Fusion Proteins; C461599:maltose transport system, E coli; D019883:Alkyl and Aryl Transferases; D051231:Farnesyltranstransferase