Isolation and evaluation of stress tolerance in acetic acid-tolerant yeasts
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Abstract
Bioethanol production from lignocellulosic materials has attracted a significant amount of interest. However, a pretreatment process of the lignocellulosic materials is needed before fermentation. This process releases microbial inhibitors, particularly acetic acid which is toxic to fermenting microorganisms and reduces ethanol yields. The aim of this study was to isolate acetic acid-tolerant yeasts obtained from natural samples. Forty-three yeast strains were isolated. The obtained isolates were then examined for their acetic acid tolerance ability by spotting a ten-fold serial dilution on YPD agar supplemented with 0.2%, 0.4%, 0.6%, 0.8%, 1.0% and 1.2% (v/v) acetic acid using Saccharomyces cerevisiae BY4743 as a comparator. The results showed that eight isolated yeasts could tolerate acetic acid up to 0.8% (v/v). However, one isolated yeast sample was able to grow in 1.0% (v/v) acetic acid and this was MY2/P1. Eight isolates were then selected for testing their ability to tolerate high temperatures and high concentrations of glucose and ethanol. It was found that L/A1 and S/PA1 could grow well at 42°C. Moreover, MY1/P3, S/PA1 and L/A1 were found to be able to tolerate glucose at 45% (w/v) and L/A1 and S/PA1 were found to tolerate ethanol at 12.5% (v/v). From this research, MY2/P1, L/A1, S/PA1 and MY1/P3 were found to possess the best properties in terms of acetic acid tolerance and the ability to grow under conditions involving high concentrations of glucose and ethanol. These strains were determined through the sequencing of the D1/D2 region of the large subunit rRNA gene (LSU) to represent to Pichia manshurica MY2/P1, P. kudriavzevii L/A1, P. kudriavzevii S/PA1 and Starmerella bacillaris MY1/P3, respectively.
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