Static microplate fermentation and automated growth analysis approaches identified a highly-aldehyde resistant Saccharomyces cerevisiae strain

• Published in: Biomass & bioenergy Vol. 120; pp. 49 - 58
• Main Authors: de Mello, Fellipe da Silveira Bezerra, Coradini, Alessandro Luis Venega, Tizei, Pedro Augusto Galvão, Carazzolle, Marcelo Falsarella, Pereira, Gonçalo Amarante Guimarães, Teixeira, Gleidson Silva
• Format: Journal Article
• Language: English
• Published: Elsevier Ltd 01-01-2019
• Subjects: High-throughput screening; Oxidative stress; Saccharomyces cerevisiae; High-throughput screening; Oxidative stress; Saccharomyces cerevisiae
• ISSN: 0961-9534; 1873-2909
• DOI: 10.1016/j.biombioe.2018.10.019

The need to replace fossil fuels has motivated research to develop alternative technologies for renewable energy production. For second generation ethanol, this includes the high-throughput screening and selection of yeast strains resistant to inhibitory byproducts of biomass pretreatment. In this work, we evaluated the use of static growth in 96-well microplates as a fermentative assay for Saccharomyces cerevisiae high-throughput screening, together with robust OD measurement correlations that overcome the low absorbance limit found in conventional plate spectrophotometers. This resulted in a method of yeast cultivation in microplates that requires no agitation and results in accurate kinetic parameters that can be used for the screening of S. cerevisiae. In order to automate calculation of these physiological parameters, we created OCHT®, an open-source high-throughput phenotype profiling software for processing, analyzing and visualizing S. cerevisiae growth data. Together, the cultivation standardization and data analysis toolbox allowed the evaluation and comparison of four S. cerevisiae strains in media containing aldehydes growth inhibitors, typically found in hydrolysates produced from sugarcane biomass. We identified FMY001 (SA1-derived) as a tolerant strain at high concentrations of HMF, furfural and vanillin. The individual analysis of segregants derived from FMY001 demonstrated their potential application in emerging biotechnological processes and gave us preliminary insights about the genetic mechanisms related to its high HMF concentration tolerance.
•Static growth and OD correction in microplate cultivations result in accurate kinetic growth parameters of S. cerevisiae.•OCHT®, an open-source MATLAB® software, allows simple high-throughput analysis of yeast growth data.•The cultivation and data analysis aproaches allowed the selection of an aldehyde highly-resistant yeast strain, FMY001.•Analysis of FMY001’s segregants gives insight into the genetic mechanisms related to its HMF high concentration tolerance.