Molecular cloning, heterologous expression, and functional characterization of a cellulolytic enzyme (Cel PRII) from buffalo rumen metagenome.

A cellulase encoding gene, Cel PRII, was identified from Mehsani buffalo rumen metagenome, and cloned and expressed in Escherichia coli BL21(DE3)pLysS. The 1170 bp full length gene encodes a 389 residue polypeptide (Cel PRII) containing a catalytic domain belonging to glycosyl hydrolase (GH) 5 family. The fusion protein consisting of the Cel PRII, thioredoxin tag and 6x Histidine tag with predicted molecular weight of 63 kDa when recovered from inclusion bodies under denaturing conditions, exhibited cellulolytic activity against carboxymethyl cellulose (CMC).

Isolation and characterization of novel multifunctional recombinant family 26 glycoside hydrolase from Mehsani buffalo rumen metagenome.

Rumen microbiota harbor a diverse set of carbohydrate-active enzymes (CAZymes), which play a crucial role in the degradation of a complex plant polysaccharide thereby providing metabolic energy to the host animals. Earlier, we reported CAZYme analysis from the buffalo rumen metagenome by high throughput shotgun sequencing. Among the various CAZymes, glycoside hydrolase family 26 (GH26) enzymes have a number of industrial applications including in paper, oil, biofuel, food, feed, pharmaceutical, coffee, and detergent industries.

Goat activin receptor type IIB knockdown by artificial microRNAs in vitro.

Activin receptor type IIB (ACVR2B) has been known to negatively regulate the muscle growth through mediating the action of transforming growth factor beta superfamily ligands. Recently, the artificial microRNAs (amiRNAs) which are processed by endogenous miRNA processing machinery have been proposed as promising approach for efficient gene knockdown. We evaluated amiRNAs targeting goat ACVR2B in HEK293T and goat myoblasts cells.