Expression and export of Pseudomonas putida NTU-8 creatinase by Escherichia coli using the chitinase signal sequence of Aeromonas hydrophila.
The gene for the creatinase from Pseudomonas putida NTU-8 was sequenced and revealed an open reading frame (ORF) of 1209 base pairs encoding a polypeptide of 403 amino acids with a calculated molecular weight (M(r)) of 45,691. The deduced amino acid sequence is very similar to that of the creatinase of Pseudomonas putida and Flavobacterium sp. An overproduction system for the chitinase signal peptide--creatinase hybrid gene was constructed by using the pQE-51 expression vector in E. coli JM109. The amount of this fusion enzyme was about 50% exported into the periplasmic space of E. coli.
Hong
M C
MC
Department of Biochemistry, Medical College, National Cheng Kung University, Tainan, Taiwan, ROC.
Chang
J C
JC
Wu
M L
ML
Chang
M C
MC
D016980:Aeromonas hydrophila / Q000201:enzymology*; D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport; D002688:Chitinases / Q000737:chemistry*; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000235:genetics; D016366:Open Reading Frames; D021382:Protein Sorting Signals / Q000737:chemistry*; D016958:Pseudomonas putida / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid; D014512:Ureohydrolases / Q000235:genetics* / Q000378:metabolism
D004269:DNA, Bacterial; D021382:Protein Sorting Signals; D002688:Chitinases; D014512:Ureohydrolases; C019944:creatinase