Cloning and Characterization of a Novel ��-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.

Strain improvement studies on <i>Microbacterium foliorum</i> GA2 for production of α-amylase in solid state fermentation: Biochemical characteristics and wash performance analysis at low temperatures

“…3). On the contrary, Xu et al (2014) reported that the optimum temperature for the best amylase activity of purified AmyPL was 50∞C. Thus, it can be assumed that mutation might improve the activity by altering the dynamic properties of residues in the active site of the enzyme.…”

Bacterial and Archaeal α-Amylases: Diversity and Amelioration of the Desirable Characteristics for Industrial Applications

“…Its temperature and pH optima were 10–15°C and pH 8–9. α-Amylases have also been reported from metagenomic libraries from pygmy loris ( Nycticebus pygmaeus ) and cow dung (Xu et al, 2014 ; Sharma et al, 2015 ).…”

Molecular and Biochemical Characterization of Salt-Tolerant Trehalose-6-Phosphate Hydrolases Identified by Screening and Sequencing Salt-Tolerant Clones From the Metagenomic Library of the Gastrointestinal Tract

“…The animal is believed to be domesticated for as long as 8000 years, during which common salt was exclusively provided to the animal by local farmers as an additional feed ( Das et al, 2011 ). This study screened salt-tolerant clones from the constructed fecal metagenomic library of N. pygmaeus and B. frontalis ( Xu et al, 2014 ; Dong et al, 2016 ). We analyzed potential salt-tolerant genes and enzymes through high-throughput sequencing and bioinformatics technology.…”