Pipeline for Targeted Meta-Proteomic Analyses to Assess the Diversity of Cattle Rumen Microbial Urease.

ABSTRACT: In the rumen of cattle, urease produced by ureolytic bacteria catalyzes the hydrolysis of urea to ammonia, which plays an important role in nitrogen metabolism and animal production. A high diversity of rumen bacterial urease genes was observed in our previous study; however, information on urease protein diversity could not be determined due to technical limitations. Here, we developed a targeted meta-proteomic pipeline to analyze rumen urease protein diversity. Protein extraction (duration of cryomilling in liquid nitrogen), protein digestion state (in-solution or in-gel), and the digestion enzyme used (trypsin or Glu-C/Lys-C) were optimized, and the digested peptides were analyzed by LC-MS/MS. Four minutes was the best duration for cryomilling and yielded the highest urease activity. Tr

SUBMITTER: Zhang X 

PROVIDER: S-EPMC7531017 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature