Chitinase B from Serratia marcescens BJL200 is exported to the periplasm without processing.
A gene encoding a chitinase from Serratia marcescens BJL200 was cloned and expressed in Escherichia coli and S. marcescens. Nucleotide sequencing revealed an open reading frame encoding a 55.5 kDa protein of 499 amino acids without a typical signal peptide for export. The cellular localization of the chitinase was studied, using two types of cell fractionation methods of immunocytochemical techniques. These analyses showed that the chitinase is located in the cytoplasm in E. coli, whereas it is exported to the periplasm in S. marcescens. Analysis of chitinase isolated from periplasmic fractions of S. marcescens carrying the cloned gene showed that export of the enzyme is not accompanied by processing at the N-terminus. The chitinase did not show any of the characteristics that have been proposed to direct the export of other non-processed extracellular proteins such as the E. coli haemolysin and might therefore be secreted via a hitherto unknown mechanism.
Brurberg
M B
MB
Laboratory of Microbial Gene Technology, Agricultural University of Norway, As.
Eijsink
V G
VG
Haandrikman
A J
AJ
Venema
G
G
Nes
I F
IF
D000595:Amino Acid Sequence; D001483:Base Sequence; D002688:Chitinases / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000648:ultrastructure; D015870:Gene Expression; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D015183:Restriction Mapping; D012706:Serratia marcescens / Q000201:enzymology* / Q000235:genetics
D004269:DNA, Bacterial; D011994:Recombinant Proteins; D002688:Chitinases