Title: Targeted quantification of functional enzyme dynamics in environmental samples for microbially mediated biogeochemical processes

Abstract

Summary Microbial enzymes catalytically drive biogeochemical processes in environments. The dynamic linkage between functional enzymes and biogeochemical species transformation has, however, rarely been investigated for decades because of the challenges to directly quantify enzymes in environmental samples. The diversity of microorganisms, the low amount of available biomass and the complexity of chemical composition in environmental samples represent the main challenges. To address the diversity challenge, we first identify several signature peptides that are conserved in the targeted enzymes with the same functionality across many phylogenetically diverse microorganisms using metagenome‐based protein sequence data. Quantification of the signature peptides then allows estimation of the targeted enzyme abundance. To achieve analyses of the requisite sensitivity for complex environmental samples with low available biomass, we adapted a recently developed ultrasensitive targeted quantification technology, termed high‐pressure high‐resolution separations with intelligent selection and multiplexing (PRISM) by improving peptide separation efficiency and method detection sensitivity. Nitrate reduction dynamics catalyzed by dissimilatory and assimilatory enzymes in a hyporheic zone sediment was used as an example to demonstrate the application of the enzyme quantification approach. Together with the measurements of biogeochemical species, the approach enables investigating the dynamic linkage between functional enzymes and biogeochemical processes.

Authors:

Li, Minjing ^[1]; Gao, Yuqian ^[2]; Qian, Wei‐Jun ^[2]; Shi, Liang ^[2]; Liu, Yuanyuan ^[2]; Nelson, William C. ^[2]; Nicora, Carrie D. ^[2]; Resch, Charles T. ^[2]; Thompson, Christopher ^[2]; Yan, Sen ^[1]; Fredrickson, James K. ^[2]; Zachara, John M. ^[2]; Liu, Chongxuan ^[3]

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+ Show Author Affiliations

1. School of Environmental Studies China University of Geosciences Wuhan 430074 People's Republic of China
2. Pacific Northwest National Laboratory Richland, WA 99354 USA
3. Pacific Northwest National Laboratory Richland, WA 99354 USA, School of Environmental Science and Engineering Southern University of Science and Technology Shenzhen 518055 People's Republic of China

Publication Date:

Thu Jul 13 00:00:00 EDT 2017

Sponsoring Org.:

USDOE

OSTI Identifier:

1399275

Resource Type:

Publisher's Accepted Manuscript

Journal Name:

Environmental Microbiology Reports

Additional Journal Information:

Journal Name: Environmental Microbiology Reports Journal Volume: 9 Journal Issue: 5; Journal ID: ISSN 1758-2229

Publisher:

Wiley-Blackwell

Country of Publication:

United States

Language:

English