Metaproteomics-guided selection of targeted enzymes for bioprospecting of mixed microbial communities

Biotechnology for Biofuels
Jutta SpedaMartin Karlsson

Abstract

Hitherto, the main goal of metaproteomic analyses has been to characterize the functional role of particular microorganisms in the microbial ecology of various microbial communities. Recently, it has been suggested that metaproteomics could be used for bioprospecting microbial communities to query for the most active enzymes to improve the selection process of industrially relevant enzymes. In the present study, to reduce the complexity of metaproteomic samples for targeted bioprospecting of novel enzymes, a microbial community capable of producing cellulases was maintained on a chemically defined medium in an enzyme suppressed metabolic steady state. From this state, it was possible to specifically and distinctively induce the desired cellulolytic activity. The extracellular fraction of the protein complement of the induced sample could thereby be purified and compared to a non-induced sample of the same community by differential gel electrophoresis to discriminate between constitutively expressed proteins and proteins upregulated in response to the inducing substance. Using the applied approach, downstream analysis by mass spectrometry could be limited to only proteins recognized as upregulated in the cellulase-induced sample...Continue Reading

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Citations

Oct 14, 2017·FEMS Microbiology Letters·Francesca BeriniFlavia Marinelli
Jan 14, 2021·Bioresource Technology·Shashi Kant BhatiaGopalakrishnan Kumar
Oct 9, 2019·Current Opinion in Biotechnology·Stephen P LillingtonMichelle A O'Malley
Mar 31, 2021·Annual Review of Biomedical Engineering·Patrick A LeggieriOphelia S Venturelli

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Methods Mentioned

BETA
electrophoresis
reverse phase chromatography
acetylation
PCR

Software Mentioned

PEAKS DB
ExPASy
SPIDER
PEAKS
SPIDER SPIDER
PEAKS SPIDER
Compute pI / M w
Phobius
PEAKS DB SPIDER
PDQuest

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