The influence of pressure and treatment time on cells disruption of different black yeasts and on activities of extracted proteins using supercritical carbon dioxide process was studied. The cells of three different black yeasts Phaeotheca triangularis, Trimatostroma salinum, and Wallemia ichthyophaga were exposed to supercritical carbon dioxide (SC CO2) by varying pressure at fixed temperature (35°C). The black yeasts cell walls were disrupted, and the content of the cells was spilled into the liquid medium. The impact of SC CO2 conditions on secretion of enzymes and proteins from black yeast cells suspension was studied. The residual activity of the enzymes cellulase, β-glucosidase, α-amylase, and protease was studied by enzymatic assay. The viability of black yeast cells was determined by measuring the optical density of the cell suspension at 600 nm. The total protein concentration in the suspension was determined on UV–Vis spectrophotometer at 595 nm. The release of intracellular and extracellular products from black yeast cells was achieved. Also, the observation by an environmental scanning electron microscopy shows major morphological changes with SC CO2-treated cells. The advantages of the proposed method are in a simple use, which is also possible for heat-sensitive materials on one hand and on the other hand integration of the extraction of enzymes and their use in biocatalytical reactions.
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机译:研究了压力和处理时间对不同黑酵母细胞分裂的影响以及对超临界二氧化碳法提取蛋白活性的影响。通过在固定温度(35°C)下改变压力,将三种不同的黑色酵母Phaeotheca triangleis,Trimatostroma salinum和Wallemia ichthyophaga的细胞暴露于超临界二氧化碳(SC CO2)。黑酵母细胞壁被破坏,并且细胞的内容物溢出到液体培养基中。研究了SC CO2条件对黑酵母细胞悬液中酶和蛋白质分泌的影响。通过酶促分析研究了纤维素酶,β-葡萄糖苷酶,α-淀粉酶和蛋白酶的残留活性。黑酵母细胞的生存力是通过测量细胞悬液在600nm的光密度来确定的。用紫外可见分光光度计在595 nm处测定悬浮液中的总蛋白质浓度。实现了从黑酵母细胞释放细胞内和细胞外产物。而且,通过环境扫描电子显微镜的观察显示了SC CO 2处理的细胞的主要形态变化。所提出的方法的优点是使用简单,一方面对于热敏材料也是可能的,另一方面,酶提取的整合及其在生物催化反应中的应用也是可能的。
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