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

Autor(es)

Temer, Beatriz ; dos Santos, Leandro Vieira ; Negri, Victor Augusti ; Galhardo, Juliana Pimentel ; Mello Magalhaes, Pedro Henrique ; Jose, Juliana ; Marschalk, Cidnei ; Ribeiro Correa, Thamy Livia ; Carazzolle, Marcelo Falsarella ; Guimaraes Pereira, Goncalo Amarante

• Tipo de documento: Artigo Científico
• Fonte: BMC Biotechnology; v. 17, SEP 9 2017.
• Citações Web of Science: 6

Resumo

Background: 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. Results: 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. Conclusions: 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. (AU)

• Processo FAPESP: 12/05827-8 - Caracterização da xilose isomerase de Propionibacterium acidipropionici e expressão em Saccharomyces cerevisiae visando a produção de etanol de segunda geração
• Beneficiário: Beatriz Temer
• Modalidade de apoio: Bolsas no Brasil - Mestrado