Characterization of a chitinase gene from Stenotrophomonas maltophilia strain 34S1 and its involvement in biological control.
A chitinase gene was cloned on a 2.8-kb DNA fragment from Stenotrophomonas maltophilia strain 34S1 by heterologous expression in Burkholderia cepacia. Sequence analysis of this fragment identified an open reading frame encoding a deduced protein of 700 amino acids. Removal of the signal peptide sequence resulted in a predicted protein that was 68 kDa in size. Analysis of the sequence indicated that the chitinase contained a catalytic domain belonging to family 18 of glycosyl hydrolases. Three putative binding domains, a chitin binding domain, a novel polycystic kidney disease (PKD) domain, and a fibronectin type III domain, were also identified within the sequence. Pairwise comparisons of each domain to the most closely related sequences found in database searches clearly demonstrated variation in gene sources and the species from which related sequences originated. A 51-kDa protein with chitinolytic activity was purified from culture filtrates of S. maltophilia strain 34S1 by hydrophobic interaction chromatography. Although the protein was significantly smaller than the size predicted from the sequence, the N-terminal sequence verified that the first 15 amino acids were identical to the deduced sequence of the mature protein encoded by chiA. Marker exchange mutagenesis of chiA resulted in mutant strain C5, which was devoid of chitinolytic activity and lacked the 51-kDa protein in culture filtrates. Strain C5 was also reduced in the ability to suppress summer patch disease on Kentucky bluegrass, supporting a role for the enzyme in the biocontrol activity of S. maltophilia.
Kobayashi
Donald Y
DY
Department of Plant Biology & Pathology, Cook College, Rutgers State University, New Brunswick, New Jersey 08901, USA. Kobayashi@aesop.rutgers.edu
Reedy
Ralph M
RM
Bick
JulieAnn
J
Oudemans
Peter V
PV
D000595:Amino Acid Sequence; D016956:Burkholderia cepacia / Q000201:enzymology / Q000235:genetics; D002686:Chitin / Q000378:metabolism; D002688:Chitinases / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D020082:Magnaporthe / Q000254:growth & development; D008969:Molecular Sequence Data; D010572:Pest Control, Biological; D010935:Plant Diseases / Q000382:microbiology; D006109:Poaceae / Q000382:microbiology*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D020615:Stenotrophomonas maltophilia / Q000201:enzymology* / Q000235:genetics
D002686:Chitin; D002688:Chitinases