title: A new locus affects cell motility, cellulose binding, and degradation by Cytophaga hutchinsonii.
abstract: Cytophaga hutchinsonii is a Gram-negative gliding bacterium, which can rapidly degrade crystalline cellulose via a novel strategy without any recognizable processive cellulases. Its mechanism of cellulose binding and degradation is still a mystery. In this study, the mutagenesis of C. hutchinsonii with the mariner-based transposon HimarEm3 and gene complementation with the oriC-based plasmid carrying the antibiotic resistance gene cfxA or tetQ were reported for the first time to provide valuable tools for mutagenesis and genetic manipulation of the bacterium. Mutant A-4 with a transposon mutation in gene CHU_0134, which encodes a putative thiol-disulfide isomerase exhibits defects in cell motility and cellulose degradation. The cellulose binding ability of A-4 was only half of that of the wild-type strain, while the endo-cellulase activity of the cell-free supernatants and on the intact cell surface of A-4 decreased by 40%. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of proteins binding to cellulose in the outer membrane showed that most of them were significantly decreased or disappeared in A-4 including some Gld proteins and hypothetical proteins, indicating that these proteins might play an important role in cell motility and cellulose binding and degradation by the bacterium.
authors.affiliation: State Key Laboratory of Microbial Technology, College of Life Science, Shandong University, Jinan 250100, China.
authors.forename: Xiaofei
authors.initials: X
authors.lastname: Ji
authors.forename: Yuanxi
authors.initials: Y
authors.lastname: Xu
authors.forename: Cong
authors.initials: C
authors.lastname: Zhang
authors.forename: Ning
authors.initials: N
authors.lastname: Chen
authors.forename: Xuemei
authors.initials: X
authors.lastname: Lu
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis; D002482:Cellulose / Q000378:metabolism*; D003589:Cytophaga / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D004251:DNA Transposable Elements; D004591:Electrophoresis, Polyacrylamide Gel; D055786:Gene Knockout Techniques; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D005827:Genetics, Microbial / Q000379:methods; D008124:Locomotion; D008967:Molecular Biology / Q000379:methods; D016254:Mutagenesis, Insertional; D019704:Protein Disulfide-Isomerases / Q000235:genetics; D020543:Proteome / Q000032:analysis
chemical_list: D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D020543:Proteome; D002482:Cellulose; D019704:Protein Disulfide-Isomerases