Cooperative degradation of chitin by extracellular and cell surface-expressed chitinases from Paenibacillus sp. strain FPU-7.
Chitin, a major component of fungal cell walls and invertebrate cuticles, is an exceedingly abundant polysaccharide, ranking next to cellulose. Industrial demand for chitin and its degradation products as raw materials for fine chemical products is increasing. A bacterium with high chitin-decomposing activity, Paenibacillus sp. strain FPU-7, was isolated from soil by using a screening medium containing α-chitin powder. Although FPU-7 secreted several extracellular chitinases and thoroughly digested the powder, the extracellular fluid alone broke them down incompletely. Based on expression cloning and phylogenetic analysis, at least seven family 18 chitinase genes were found in the FPU-7 genome. Interestingly, the product of only one gene (chiW) was identified as possessing three S-layer homology (SLH) domains and two glycosyl hydrolase family 18 catalytic domains. Since SLH domains are known to function as anchors to the Gram-positive bacterial cell surface, ChiW was suggested to be a novel multimodular surface-expressed enzyme and to play an important role in the complete degradation of chitin. Indeed, the ChiW protein was localized on the cell surface. Each of the seven chitinase genes (chiA to chiF and chiW) was cloned and expressed in Escherichia coli cells for biochemical characterization of their products. In particular, ChiE and ChiW showed high activity for insoluble chitin. The high chitinolytic activity of strain FPU-7 and the chitinases may be useful for environmentally friendly processing of chitin in the manufacture of food and/or medicine.
Department of Bioscience, Fukui Prefectural University, Eiheiji, Fukui, Japan.
Takafumi
T
Itoh
Takao
T
Hibi
Yutaka
Y
Fujii
Ikumi
I
Sugimoto
Akihiro
A
Fujiwara
Fumiko
F
Suzuki
Yukimoto
Y
Iwasaki
Jin-Kyung
JK
Kim
Akira
A
Taketo
Hisashi
H
Kimoto
D001431:Bacteriological Techniques / Q000379:methods; D002686:Chitin / Q000235:genetics / Q000378:metabolism*; D002688:Chitinases / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D003470:Culture Media / Q000737:chemistry; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D008403:Mass Screening / Q000379:methods; D008969:Molecular Sequence Data; D056507:Paenibacillus / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology
D003470:Culture Media; D004269:DNA, Bacterial; D002686:Chitin; D002688:Chitinases