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MET17 and Hydrogen Sulfide Formation inSaccharomyces cerevisiae

机译:酿酒酵母中的MET17和硫化氢的形成

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Commercial isolates of Saccharomyces cerevisiae differ in the production of hydrogen sulfide (H2S) during fermentation, which has been attributed to variation in the ability to incorporate reduced sulfur into organic compounds. We transformed two commercial strains (UCD522 and UCD713) with a plasmid overexpressing the MET17 gene, which encodes the bifunctionalO-acetylserine/O-acetylhomoserine sulfhydrylase (OAS/OAH SHLase), to test the hypothesis that the level of activity of this enzyme limits reduced sulfur incorporation, leading to H2S release. Overexpression of MET17 resulted in a 10- to 70-fold increase in OAS/OAH SHLase activity in UCD522 but had no impact on the level of H2S produced. In contrast, OAS/OAH SHLase activity was not as highly expressed in transformants of UCD713 (0.5- to 10-fold) but resulted in greatly reduced H2S formation. Overexpression of OAS/OAH SHLase activity was greater in UCD713 when grown under low-nitrogen conditions, but the impact on reduction of H2S was greater under high-nitrogen conditions. Thus, there was not a good correlation between the level of enzyme activity and H2S production. We measured cellular levels of cysteine to determine the impact of overexpression of OAS/OAH SHLase activity on sulfur incorporation. While Met17p activity was not correlated with increased cysteine production, conditions that led to elevated cytoplasmic levels of cysteine also reduced H2S formation. Our data do not support the simple hypothesis that variation in OAS/OAH SHLase activity is correlated with H2S production and release.
机译:啤酒酵母的商业分离物在发酵过程中硫化氢(H 2 S)的产生有所不同,这归因于将还原硫结合到有机化合物中的能力的差异。 。我们用过表达 MET17 基因的质粒转化了两个商业菌株(UCD522和UCD713),该基因编码双功能 O -乙酰丝氨酸/ O -乙酰高丝氨酸巯基化酶(OAS / OAH SHLase),以检验以下假设:该酶的活性水平限制了硫的掺入,导致H 2 S释放。 MET17 的过表达导致UCD522中OAS / OAH SHLase活性增加10到70倍,但对产生的H 2 S水平没有影响。相反,OAS / OAH SHLase活性在UCD713的转化子中表达不高(0.5到10倍),但导致H 2 S的形成大大减少。在低氮条件下生长时,UCD713中OAS / OAH SHLase的过表达更大,而在高氮条件下对H 2 S还原的影响更大。因此,酶活性水平与H 2 S的产生之间没有很好的相关性。我们测量了半胱氨酸的细胞水平,以确定过表达的OAS / OAH SHLase活性对硫掺入的影响。尽管Met17p活性与半胱氨酸产量增加无关,但导致半胱氨酸胞质水平升高的条件也减少了H 2 S的形成。我们的数据不支持简单的假设,即OAS / OAH SHLase活性的变化与H 2 S的产生和释放相关。

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