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Use of supercritical carbon dioxide for proteins and alcohol dehydrogenase release from yeast Saccharomyces cerevisiae

机译:使用超临界二氧化碳从酵母中释放蛋白质和乙醇脱氢酶

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In this study, supercritical carbon dioxide (SC CO2) was used to obtain proteins and enzymes from yeast Saccharomyces cerevisiae.The changes in viability and cell morphology, the release of the cellular proteins and the activity alteration of the enzyme alcohol dehydrogenase (ADH) from S. cerevisiae resulting from the exposure to SC CO2 were investigated. Before the treatment of S. cerevisiae in SC CO2, the number of viable cells was cca. 10~5 colony forming units per mL of cell suspension (cfu/mL). The suspension of the S. cerevisiae culture was incubated in SC CO2 at different pressures (7.5,15 and 30MPa) for different treatment times (30-300 min) and at constant temperature (35 ℃). The influences of these parameters on total protein concentration, ADH activity and changes in absorbance of nucleic acids (NA) in the suspension of S. cerevisiae during SC CO2 treatment were studied. A decrease in number of living cells of S. cerevisiae in potato dextrose broth (PDB) or in sodium pyrophosphate buffer (SPB) exposed to SC CO2 at all studied pressures was determined when the treatment time was increased. The highest activity of ADH after treatment of the yeast culture suspended in PDB in SC CO2 was detected at a pressure of 7.5 MPa and at the treatment time of 120 min.The use of SC CO2 is a suitable option to achieve cell death and consequently the secretion of proteins and ADH from cells of S. cerevisiae.
机译:本研究使用超临界二氧化碳(SC CO2)从酿酒酵母(Saccharomyces cerevisiae)中获得蛋白质和酶,并从酵母中获得活力和细胞形态的变化,细胞蛋白的释放以及乙醇脱氢酶(ADH)的活性变化。研究了因暴露于SC CO2而导致的酿酒酵母。在SC CO2中处理酿酒酵母之前,活细胞数为cca。每毫升细胞悬液10〜5个菌落形成单位(cfu / mL)。将酿酒酵母培养物的悬浮液在SC CO2中于不同压力(7.5、15和30MPa)下孵育不同的处理时间(30-300分钟),并在恒定温度(35℃)下孵育。研究了这些参数对SC CO2处理过程中酿酒酵母悬浮液中总蛋白浓度,ADH活性和核酸(NA)吸光度变化的影响。当增加处理时间时,确定了在所有研究压力下暴露于SC CO2的马铃薯葡萄糖肉汤(PDB)或焦磷酸钠缓冲液(SPB)中酿酒酵母活细胞数量的减少。在7.5 MPa的压力和120分钟的处理时间下,处理悬浮在PDB中的SCCO2中的酵母培养物后,ADH的活性最高。使用SC CO2是实现细胞死亡的合适选择,因此可以使细胞死亡。从酿酒酵母细胞分泌蛋白质和ADH。

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