ArticleACS chemical biology2014
Phylogenomically guided identification of industrially relevant GH1 β-glucosidases through DNA synthesis and nanostructure-initiator mass spectrometry.
Article in ACS chemical biology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed, 62 citations in OpenAlex.
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- Genomic insights into biosynthesis and adaptation in the bioactive marine bacterium Streptomyces albidoflavus VIP-1 from the Red Sea.BMC microbiology · 2025Article
- Discovery of Potent Glycosidases Enables Quantification of Smoke-Derived Phenolic Glycosides through Enzymatic Hydrolysis.Journal of agricultural and food chemistry · 2024Article
- EngineeringACS synthetic biology · 2024Article
- A glance at the gut microbiota and the functional roles of the microbes based on marmot fecal samples.Frontiers in microbiology · 2023Article
- Azido Groups Hamper Glycan Acceptance by Carbohydrate Processing Enzymes.ACS central science · 2022Article
- A multiplexed nanostructure-initiator mass spectrometry (NIMS) assay for simultaneously detecting glycosyl hydrolase and lignin modifying enzyme activities.Scientific reports · 2021Article
- High Potential for Biomass-Degrading Enzymes Revealed by Hot Spring Metagenomics.Frontiers in microbiology · 2021Article
- A glucotolerant β-glucosidase from the fungus Talaromyces amestolkiae and its conversion into a glycosynthase for glycosylation of phenolic compounds.Microbial cell factories · 2020Review
- Building a custom high-throughput platform at the Joint Genome Institute for DNA construct design and assembly-present and future challenges.Synthetic biology (Oxford, England) · 2020Article
- Engineered Root Bacteria Release Plant-Available Phosphate from Phytate.Applied and environmental microbiology · 2019Article
- A High-Throughput Mass Spectrometric Enzyme Activity Assay Enabling the Discovery of Cytochrome P450 Biocatalysts.Angewandte Chemie (International ed. in English) · 2019Article
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- A combinatorial approach to synthetic transcription factor-promoter combinations for yeast strain engineering.Yeast (Chichester, England) · 2018Article
- Metagenomic Analysis of Bacteria, Fungi, Bacteriophages, and Helminths in the Gut of Giant Pandas.Frontiers in microbiology · 2018Article
- Profiling of Microbial Colonies for High-Throughput Engineering of Multistep Enzymatic Reactions via Optically Guided Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry.Journal of the American Chemical Society · 2017Article
- Biofuel metabolic engineering with biosensors.Current opinion in chemical biology · 2016Review
- Exploiting members of the BAHD acyltransferase family to synthesize multiple hydroxycinnamate and benzoate conjugates in yeast.Microbial cell factories · 2016Article
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Authors and funding
28 authors at 5 institutions in 1 country.
Funding
Abstract
Harnessing the biotechnological potential of the large number of proteins available in sequence databases requires scalable methods for functional characterization. Here we propose a workflow to address this challenge by combining phylogenomic guided DNA synthesis with high-throughput mass spectrometry and apply it to the systematic characterization of GH1 β-glucosidases, a family of enzymes necessary for biomass hydrolysis, an important step in the conversion of lignocellulosic feedstocks to fuels and chemicals. We synthesized and expressed 175 GH1s, selected from over 2000 candidate sequences to cover maximum sequence diversity. These enzymes were functionally characterized over a range of temperatures and pHs using nanostructure-initiator mass spectrometry (NIMS), generating over 10,000 data points. When combined with HPLC-based sugar profiling, we observed GH1 enzymes active over a broad temperature range and toward many different β-linked disaccharides. For some GH1s we also observed activity toward laminarin, a more complex oligosaccharide present as a major component of macroalgae. An area of particular interest was the identification of GH1 enzymes compatible with the ionic liquid 1-ethyl-3-methylimidazolium acetate ([C2mim][OAc]), a next-generation biomass pretreatment technology. We thus searched for GH1 enzymes active at 70 °C and 20% (v/v) [C2mim][OAc] over the course of a 24-h saccharification reaction. Using our unbiased approach, we identified multiple enzymes of different phylogentic origin with such activities. Our approach of characterizing sequence diversity through targeted gene synthesis coupled to high-throughput screening technologies is a broadly applicable paradigm for a wide range of biological problems.
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