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首页> 外文期刊>New biotechnology >CTAB, Triton X-100 and freezing–thawing treatments of Candida guilliermondii: Effects on permeability and accessibility of the glucose-6- phosphate dehydrogenase, xylose reductase and xylitol dehydrogenase enzymes
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CTAB, Triton X-100 and freezing–thawing treatments of Candida guilliermondii: Effects on permeability and accessibility of the glucose-6- phosphate dehydrogenase, xylose reductase and xylitol dehydrogenase enzymes

机译：CTAB，Triton X-100和古德氏假丝酵母的冻融处理：对葡萄糖-6-磷酸脱氢酶，木糖还原酶和木糖醇脱氢酶的渗透性和可及性的影响

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Cells of Candida guilliermondii (ATCC 201935) were permeabilised with surfactant treatment (CTAB or Triton X-100) or a freezing–thawing procedure. Treatments were monitored by in situ activities of the key enzymes involved in xylose metabolism, that is, glucose-6-phosphate dehydrogenase (G6PD), xylose reductase (XR) and xylitol dehydrogenase (XD). The permeabilising ability of the surfactants was dependent on its concentration and incubation time. The optimum operation conditions for the permeabilisation of C. guilliermondii with surfactants were 0.41 mM (CTAB) or 2.78 mM (Triton X-100), 30 8C, and pH 7 at 200 rpm for 50 min. The maximum permeabilisation measured in terms of the in situ G6PD activity observed was, in order, as follows: CTAB (122.4±15.7 U/g_(cells)) > freezing–thawing (54.3±1.9 U/g_(cells)) > Triton X-100 (23.5±0.0 U/gcells). These results suggest that CTAB surfactant is more effective in the permeabilisation of C. guilliermondii cells in comparison to the freezing–thawing and Triton X-100 treatments. Nevertheless, freezing–thawing was the only treatment that allowed measurable in situ XR activity. Therefore, freezing–thawing permeabilised yeast cells could be used as a source of xylose reductase for analytical purposes or for use in biotransformation process such as xylitol preparation from xylose. The level of in situ xylose reductase was found to be 13.2±0.1 U/gcells.
机译：用表面活性剂处理（CTAB或Triton X-100）或冻融程序透化古德念珠菌细胞（ATCC 201935）。通过涉及木糖代谢的关键酶的原位活性监测治疗，所述关键酶即葡萄糖-6-磷酸脱氢酶（G6PD），木糖还原酶（XR）和木糖醇脱氢酶（XD）。表面活性剂的透化能力取决于其浓度和孵育时间。用表面活性剂透化古氏梭状芽胞杆菌的最佳操作条件是0.41 mM（CTAB）或2.78 mM（Triton X-100），30 8C和pH 7在200 rpm下持续50分钟。根据观察到的原位G6PD活性测得的最大通透性依次为：CTAB（122.4±15.7 U / g_（细胞））>冻融（54.3±1.9 U / g_（细胞））> Triton X-100（23.5±0.0 U / gcells）。这些结果表明，与冷冻-解冻和Triton X-100处理相比，CTAB表面活性剂在圭亚那梭菌细胞的透化中更有效。但是，冻融是唯一可以测量原位XR活性的方法。因此，冻融透化的酵母细胞可以用作木糖还原酶的来源，用于分析目的或用于生物转化过程，例如从木糖制备木糖醇。发现原位木糖还原酶的水平为13.2±0.1U / g细胞。

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