Linderholm Angela L, Findleton Carrie L, Kumar Gagandeep, Hong Yeun, Bisson Linda F
Department of Viticulture and Enology, University of California, Davis, California 95616-8749, USA.
Appl Environ Microbiol. 2008 Mar;74(5):1418-27. doi: 10.1128/AEM.01758-07. Epub 2008 Jan 11.
A screen of the Saccharomyces cerevisiae deletion strain set was performed to identify genes affecting hydrogen sulfide (H(2)S) production. Mutants were screened using two assays: colony color on BiGGY agar, which detects the basal level of sulfite reductase activity, and production of H(2)S in a synthetic juice medium using lead acetate detection of free sulfide in the headspace. A total of 88 mutants produced darker colony colors than the parental strain, and 4 produced colonies significantly lighter in color. There was no correlation between the appearance of a dark colony color on BiGGY agar and H(2)S production in synthetic juice media. Sixteen null mutations were identified as leading to the production of increased levels of H(2)S in synthetic juice using the headspace analysis assay. All 16 mutants also produced H(2)S in actual juices. Five of these genes encode proteins involved in sulfur containing amino acid or precursor biosynthesis and are directly associated with the sulfate assimilation pathway. The remaining genes encode proteins involved in a variety of cellular activities, including cell membrane integrity, cell energy regulation and balance, or other metabolic functions. The levels of hydrogen sulfide production of each of the 16 strains varied in response to nutritional conditions. In most cases, creation of multiple deletions of the 16 mutations in the same strain did not lead to a further increase in H(2)S production, instead often resulting in decreased levels.
为了鉴定影响硫化氢(H₂S)产生的基因,对酿酒酵母缺失菌株集进行了筛选。使用两种检测方法筛选突变体:在BiGGY琼脂上的菌落颜色,用于检测亚硫酸盐还原酶活性的基础水平;以及在合成果汁培养基中使用醋酸铅检测顶空中游离硫化物来检测H₂S的产生。共有88个突变体产生的菌落颜色比亲本菌株更深,4个产生的菌落颜色明显更浅。在BiGGY琼脂上出现深色菌落颜色与在合成果汁培养基中产生H₂S之间没有相关性。使用顶空分析检测方法,鉴定出16个无效突变会导致在合成果汁中产生更高水平的H₂S。所有16个突变体在实际果汁中也会产生H₂S。其中5个基因编码参与含硫氨基酸或前体生物合成的蛋白质,并且与硫酸盐同化途径直接相关。其余基因编码参与多种细胞活动的蛋白质,包括细胞膜完整性、细胞能量调节和平衡或其他代谢功能。16个菌株中每个菌株的硫化氢产生水平因营养条件而异。在大多数情况下,在同一菌株中创建16个突变的多个缺失并不会导致H₂S产生进一步增加,反而常常导致水平降低。