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KMID : 1236020190470010069
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Microbiology and Biotechnology Letters
2019 Volume.47 No. 1 p.69 ~ p.77
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A MALDI-MS-based Glucan Hydrolase Assay Method for Whole-cell Biocatalysis
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Ahn Da-Hee
Park Han-Gyu
Song Won-Suk
Kim Seong-Min
Jo Sung-Hyun
Yang Yung-Hun
Kim Yun-Gon
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Abstract
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Screening microorganisms that can produce glucan hydrolases for industrial, environmental, and biomedical applications is important. Herein, we describe a novel approach to perform glucan hydrolase screening? based on analysis of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) spectra?which involves degradation of the oligo- and polysaccharides. As a proof-of-concept study, glucan hydrolases that could break down glucans made of several glucose units were used to demonstrate the MALDI-MS-based enzyme assay. First, the enzyme activities of α-amylase and cellulase on a mixture of glucan oligosaccharides were successfully discriminated, where changes of the MALDI-MS profiles directly reflected the glucan hydrolase activities. Next, we validated that this MALDI-MS-based enzyme assay could be applied to glucan polysaccharides (i.e., pullulan, lichenan, and schizophyllan). Finally, the bacterial glucan hydrolase activities were screened on 96-well plate-based platforms, using cell lysates or samples of secreted enzyme. Our results demonstrated that the MALDI-MS-based enzyme assay system would be useful for investigating bacterial glucoside hydrolases in a high-throughput manner.
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KEYWORD
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Glucan hydrolase, glucan, oligosaccharides, polysaccharides, MALDI-MS, whole-cell biocatalysis
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