Conversion of an inactive xylose isomerase into a functional enzyme by co-expression of GroEL-GroES chaperonins in Saccharomyces cerevisiae

Second-generation ethanol production is a clean bioenergy source with potential to mitigate fossil fuel emissions. The engineering of Saccharomyces cerevisiae for xylose utilization is an essential step towards the production of this biofuel. Though xylose isomerase (XI) is the key enzyme for xylose...

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Published in:	BMC biotechnology Vol. 17; no. 1; p. 71
Main Authors:	Temer, Beatriz, Dos Santos, Leandro Vieira, Negri, Victor Augusti, Galhardo, Juliana Pimentel, Magalhães, Pedro Henrique Mello, José, Juliana, Marschalk, Cidnei, Corrêa, Thamy Lívia Ribeiro, Carazzolle, Marcelo Falsarella, Pereira, Gonçalo Amarante Guimarães
Format:	Journal Article
Language:	English
Published:	England BioMed Central Ltd 09-09-2017 BioMed Central BMC
Subjects:	Alcohol Alcohol, Denatured Aldose-Ketose Isomerases - chemistry Aldose-Ketose Isomerases - genetics Aldose-Ketose Isomerases - metabolism Amino acids Bacteria Biodiesel fuels Biofuels Biomass Brewer's yeast Chaperonin 60 - genetics Chaperonin 60 - metabolism Chaperonins Conversion E coli Escherichia coli Escherichia coli Proteins - genetics Escherichia coli Proteins - metabolism Ethanol Ethanol - metabolism Ethanol production Fossil fuels Fungi Genetic recombination GroEL-GroES chaperonins Heat-Shock Proteins - genetics Heat-Shock Proteins - metabolism Hydrophobicity Models, Molecular Propionibacterium - enzymology Propionibacterium acidipropionici Protein Conformation Protein Engineering - methods Protein folding Recombinant Proteins - genetics Recombinant Proteins - metabolism Saccharomyces cerevisiae Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae - metabolism Strains (organisms) Xylose Xylose - metabolism Xylose fermentation Xylose isomerase Yeast Yeasts Xylose fermentation GroEL-GroES chaperonins Xylose isomerase Saccharomyces cerevisiae Ethanol production
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Summary:	Second-generation ethanol production is a clean bioenergy source with potential to mitigate fossil fuel emissions. The engineering of Saccharomyces cerevisiae for xylose utilization is an essential step towards the production of this biofuel. Though xylose isomerase (XI) is the key enzyme for xylose conversion, almost half of the XI genes are not functional when expressed in S. cerevisiae. To date, protein misfolding is the most plausible hypothesis to explain this phenomenon. This study demonstrated that XI from the bacterium Propionibacterium acidipropionici becomes functional in S. cerevisiae when co-expressed with GroEL-GroES chaperonin complex from Escherichia coli. The developed strain BTY34, harboring the chaperonin complex, is able to efficiently convert xylose to ethanol with a yield of 0.44 g ethanol/g xylose. Furthermore, the BTY34 strain presents a xylose consumption rate similar to those observed for strains carrying the widely used XI from the fungus Orpinomyces sp. In addition, the tetrameric XI structure from P. acidipropionici showed an elevated number of hydrophobic amino acid residues on the surface of protein when compared to XI commonly expressed in S. cerevisiae. Based on our results, we elaborate an extensive discussion concerning the uncertainties that surround heterologous expression of xylose isomerases in S. cerevisiae. Probably, a correct folding promoted by GroEL-GroES could solve some issues regarding a limited or absent XI activity in S. cerevisiae. The strains developed in this work have promising industrial characteristics, and the designed strategy could be an interesting approach to overcome the non-functionality of bacterial protein expression in yeasts.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1472-6750 1472-6750
DOI:	10.1186/s12896-017-0389-7