title: Gene Cloning and Expression of Cellulase of Bacillus amyloliquefaciens Isolated from the Cecum of Goose.
abstract: A kind of bacteria secreting cellulase and showing probiotic attributes was isolated from the cecum of goose and identified as Bacillus amyloliquefaciens by analysis of 16S rRNA gene sequence and named as B. amyloliquefaciens S1. In vitro assays, the enzymatic activity of the strain was determined by the reducing-sugar method, and the proper culture conditions of producing cellulase and some properties of the cellulase were investigated. The cultural mixture of the bacteria had a high cellulase activity of 1.25 U/mL. In order to improve the utilization rate of the cellulase, some properties of the cellulase were studied. The best reaction pH of the enzymes was 7.0 and the optimum reaction temperature was 60°C. The enzyme was a kind of neutral cellulase that possessing strong resistance against heat and acidity. It showed high activity to absorbent cotton, soybean meal, and filter paper. Meanwhile, a gene encoding a kind of cellulase was cloned and prokaryotic expressed in Escherichia coli. The gene had 1500 bp in length, encoding a protein of 55 kDa, which was confirmed by SDS-PAGE and Western blotting. This study explored the possibility of degrading ability of bacteria with its probiotic attributes to enhance digestibility of the feed and gut health of animal. It also provided some basis for its further functional analysis and practical application as a microbial preparation for the breeding.
authors: a Center for Developmental Biology , School of Life Science, Anhui Agriculture University , Hefei , Anhui , Republic of China.
authors: Linghong
authors: L
authors: Sun
authors: a Center for Developmental Biology , School of Life Science, Anhui Agriculture University , Hefei , Anhui , Republic of China.
authors: Jiangyan
authors: J
authors: Cao
authors: a Center for Developmental Biology , School of Life Science, Anhui Agriculture University , Hefei , Anhui , Republic of China.
authors: Ying
authors: Y
authors: Liu
authors: a Center for Developmental Biology , School of Life Science, Anhui Agriculture University , Hefei , Anhui , Republic of China.
authors: Junjie
authors: J
authors: Wang
authors: a Center for Developmental Biology , School of Life Science, Anhui Agriculture University , Hefei , Anhui , Republic of China.
authors: Panpan
authors: P
authors: Guo
authors: a Center for Developmental Biology , School of Life Science, Anhui Agriculture University , Hefei , Anhui , Republic of China.
authors: Zaigui
authors: Z
authors: Wang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000069977:Bacillus amyloliquefaciens / Q000201:enzymology* / Q000235:genetics; D002432:Cecum / Q000382:microbiology; D002480:Cellulase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D004372:Ducks / Q000382:microbiology*; D005777:Geese / Q000382:microbiology*; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D006863:Hydrogen-Ion Concentration; D010802:Phylogeny; D017422:Sequence Analysis, DNA / Q000662:veterinary; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D002480:Cellulase
keywords: Bacillus amyloliquefaciens; cecum of goose; cellulase; enzyme properties; probiotic attributes