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Proteomics guided study of Saccharomyces cerevisiae for the development of consolidated bioprocessing chasses
  • +6
  • Rosana Goldbeck,
  • Manoela Martins,
  • Alberto M. M. Lopes,
  • Thomas Eng,
  • Yan Chen,
  • Jennifer W. Gin,
  • Christopher Petzold,
  • Aindrila Mukopadhyay,
  • Flávia V. Winck
Rosana Goldbeck
Universidade Estadual de Campinas Faculdade de Engenharia de Alimentos

Corresponding Author:rosana.goldbeck@gmail.com

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Manoela Martins
Universidade Estadual de Campinas Faculdade de Engenharia de Alimentos
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Alberto M. M. Lopes
Universidade Estadual de Campinas Faculdade de Engenharia de Alimentos
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Thomas Eng
E O Lawrence Berkeley National Laboratory The Biosciences Area Division of Biological Systems and Engineering
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Yan Chen
E O Lawrence Berkeley National Laboratory The Biosciences Area Division of Biological Systems and Engineering
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Jennifer W. Gin
E O Lawrence Berkeley National Laboratory The Biosciences Area Division of Biological Systems and Engineering
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Christopher Petzold
E O Lawrence Berkeley National Laboratory The Biosciences Area Division of Biological Systems and Engineering
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Aindrila Mukopadhyay
E O Lawrence Berkeley National Laboratory The Biosciences Area Division of Biological Systems and Engineering
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Flávia V. Winck
Universidade de Sao Paulo Centro de Energia Nuclear na Agricultura
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Abstract

New approaches to increase productivity and reduce costs may improve the use of lignocellulosic biomass in biorefineries, benefiting the biobased economy and reducing political and environmental instabilities from fossil sources. The deletion of MNN10 and PMT5 genes, related to N- and O-mannosylation processes, in Saccharomyces cerevisiae BY4742 was performed by CRISPR/Cas9 to evaluate the effects on the enzymatic activity of a heterologous laccase. Next, the mutant with the higher laccase activity, mnn10Δ, in concert with expression of lignocellulolytic enzymes ( GH74 endoglucanase, GH3 β-glucosidase) integrated in the Ty1 locus was used to create a strain for a consolidated bioprocess. Proteomic analysis showed that several proteins related to the synthesis of the cell wall, vesicle formation, protein trafficking, and glycosylation had changes in expression levels after deletion of the MNN10. This mutant strain also showed increased extracellular activities of recombinant CAZYmes, which could be a result of changes in N- and O-glycosylation or extracellular secretion. Active heterologous enzymes can be expressed from genes integrated into the Ty1 locus (transposon) of such industrial and laboratory S. cerevisiae chassis and provides a promising approach for the development of consolidated bioprocessing.