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Title: High throughput nanostructure-initiator mass spectrometry screening of microbial growth conditions for maximal β-glucosidase production

Abstract

Production of biofuels via enzymatic hydrolysis of complex plant polysaccharides is a subject of intense global interest. Microbial communities are known to express a wide range of enzymes necessary for the saccharification of lignocellulosic feedstocks and serve as a powerful reservoir for enzyme discovery. However, the growth temperature and conditions that yield high cellulase activity vary widely, and the throughput to identify optimal conditions has been limited by the slow handling and conventional analysis. A rapid method that uses small volumes of isolate culture to resolve specific enzyme activity is needed. In this work, a high throughput nanostructure-initiator mass spectrometry (NIMS)-based approach was developed for screening a thermophilic cellulolytic actinomycete, Thermobispora bispora, for β-glucosidase production under various growth conditions. Media that produced high β-glucosidase activity were found to be I/S + glucose or microcrystalline cellulose (MCC), Medium 84 + rolled oats, and M9TE + MCC at 45°C. Supernatants of cell cultures grown in M9TE + 1% MCC cleaved 2.5 times more substrate at 45°C than at all other temperatures. While T. bispora is reported to grow optimally at 60°C in Medium 84 + rolled oats and M9TE + 1% MCC, approximately 40% more conversion was observed at 45°C. This highmore » throughput NIMS approach may provide an important tool in discovery and characterization of enzymes from environmental microbes for industrial and biofuel applications.« less

Authors:
 [1];  [1];  [2];  [3];  [2];  [1];  [1];  [1]
  1. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  2. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Sandia National Lab. (SNL-CA), Livermore, CA (United States)
  3. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23). Biological Systems Science Division
OSTI Identifier:
1511348
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Frontiers in Microbiology
Additional Journal Information:
Journal Volume: 4; Journal Issue: DEC; Journal ID: ISSN 1664-302X
Publisher:
Frontiers Research Foundation
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; NIMS; high throughput; β-glucosidase; enzymatic activity screening; microbial communities

Citation Formats

Cheng, Xiaoliang, Hiras, Jennifer, Deng, Kai, Bowen, Benjamin, Simmons, Blake A., Adams, Paul D., Singer, Steven W., and Northen, Trent R. High throughput nanostructure-initiator mass spectrometry screening of microbial growth conditions for maximal β-glucosidase production. United States: N. p., 2013. Web. doi:10.3389/fmicb.2013.00365.
Cheng, Xiaoliang, Hiras, Jennifer, Deng, Kai, Bowen, Benjamin, Simmons, Blake A., Adams, Paul D., Singer, Steven W., & Northen, Trent R. High throughput nanostructure-initiator mass spectrometry screening of microbial growth conditions for maximal β-glucosidase production. United States. doi:10.3389/fmicb.2013.00365.
Cheng, Xiaoliang, Hiras, Jennifer, Deng, Kai, Bowen, Benjamin, Simmons, Blake A., Adams, Paul D., Singer, Steven W., and Northen, Trent R. Fri . "High throughput nanostructure-initiator mass spectrometry screening of microbial growth conditions for maximal β-glucosidase production". United States. doi:10.3389/fmicb.2013.00365. https://www.osti.gov/servlets/purl/1511348.
@article{osti_1511348,
title = {High throughput nanostructure-initiator mass spectrometry screening of microbial growth conditions for maximal β-glucosidase production},
author = {Cheng, Xiaoliang and Hiras, Jennifer and Deng, Kai and Bowen, Benjamin and Simmons, Blake A. and Adams, Paul D. and Singer, Steven W. and Northen, Trent R.},
abstractNote = {Production of biofuels via enzymatic hydrolysis of complex plant polysaccharides is a subject of intense global interest. Microbial communities are known to express a wide range of enzymes necessary for the saccharification of lignocellulosic feedstocks and serve as a powerful reservoir for enzyme discovery. However, the growth temperature and conditions that yield high cellulase activity vary widely, and the throughput to identify optimal conditions has been limited by the slow handling and conventional analysis. A rapid method that uses small volumes of isolate culture to resolve specific enzyme activity is needed. In this work, a high throughput nanostructure-initiator mass spectrometry (NIMS)-based approach was developed for screening a thermophilic cellulolytic actinomycete, Thermobispora bispora, for β-glucosidase production under various growth conditions. Media that produced high β-glucosidase activity were found to be I/S + glucose or microcrystalline cellulose (MCC), Medium 84 + rolled oats, and M9TE + MCC at 45°C. Supernatants of cell cultures grown in M9TE + 1% MCC cleaved 2.5 times more substrate at 45°C than at all other temperatures. While T. bispora is reported to grow optimally at 60°C in Medium 84 + rolled oats and M9TE + 1% MCC, approximately 40% more conversion was observed at 45°C. This high throughput NIMS approach may provide an important tool in discovery and characterization of enzymes from environmental microbes for industrial and biofuel applications.},
doi = {10.3389/fmicb.2013.00365},
journal = {Frontiers in Microbiology},
issn = {1664-302X},
number = DEC,
volume = 4,
place = {United States},
year = {2013},
month = {12}
}

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Cited by: 7 works
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