title: Cloning and characterization of a novel α-amylase from a fecal microbial metagenome.
abstract: To isolate novel and useful microbial enzymes from uncultured gastrointestinal microorganisms, a fecal microbial metagenomic library of the pygmy loris was constructed. The library was screened for amylolytic activity, and 8 of 50,000 recombinant clones showed amylolytic activity. Subcloning and sequence analysis of a positive clone led to the identification a novel gene (amyPL) coding for α-amylase. AmyPL was expressed in Escherichia coli BL21 (DE3) and the purified AmyPL was enzymatically characterized. This study is the first to report the molecular and biochemical characterization of a novel α-amylase from a gastrointestinal metagenomic library.
authors.affiliation: School of Life Science, Yunnan Normal University, Kunming 650500, China, Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Kunming 650500, China, Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment, Kunming 650500, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Xu
authors.forename: Fuya
authors.initials: F
authors.lastname: Yang
authors.forename: Caiyun
authors.initials: C
authors.lastname: Xiong
authors.forename: Junjun
authors.initials: J
authors.lastname: Li
authors.forename: Xianghua
authors.initials: X
authors.lastname: Tang
authors.forename: Junpei
authors.initials: J
authors.lastname: Zhou
authors.forename: Zhenrong
authors.initials: Z
authors.lastname: Xie
authors.forename: Junmei
authors.initials: J
authors.lastname: Ding
authors.forename: Yunjuan
authors.initials: Y
authors.lastname: Yang
authors.forename: Zunxi
authors.initials: Z
authors.lastname: Huang
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D016000:Cluster Analysis; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005243:Feces / Q000382:microbiology*; D015870:Gene Expression; D008142:Lorisidae; D054892:Metagenome; D008969:Molecular Sequence Data; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D000516:alpha-Amylases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D000516:alpha-Amylases