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Use of Yeast Spores for Microencapsulation of Enzymes

机译：酵母孢子用于酶的微囊化

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Here, we report a novel method to produce microencapsulated enzymes using Saccharomyces cerevisiae spores. In sporulating cells, soluble secreted proteins are transported to the spore wall. Previous work has shown that the spore wall is capable of retaining soluble proteins because its outer layers work as a diffusion barrier. Accordingly, a red fluorescent protein (RFP) fusion of the α-galactosidase, Mel1, expressed in spores was observed in the spore wall even after spores were subjected to a high-salt wash in the presence of detergent. In vegetative cells, however, the cell wall cannot retain the RFP fusion. Although the spore wall prevents diffusion of proteins, it is likely that smaller molecules, such as sugars, pass through it. In fact, spores can contain much higher α-galactosidase activity to digest melibiose than vegetative cells. When present in the spore wall, the enzyme acquires resistance to environmental stresses including enzymatic digestion and high temperatures. The outer layers of the spore wall are required to retain enzymes but also decrease accessibility of the substrates. However, mutants with mild spore wall defects can retain and stabilize the enzyme while still permitting access to the substrate. In addition to Mel1, we also show that spores can retain the invertase. Interestingly the encapsulated invertase has significantly lower activity toward raffinose than toward sucrose. This suggests that substrate selectivity could be altered by the encapsulation.
机译：在这里，我们报告一种使用酿酒酵母孢子生产微囊化酶的新方法。在孢子形成细胞中，可溶性分泌蛋白被转运到孢子壁。先前的工作表明，孢子壁能够保留可溶性蛋白质，因为其外层可作为扩散屏障。因此，即使在去污剂存在下对孢子进行高盐洗涤后，在孢子壁上也观察到了孢子中表达的α-半乳糖苷酶Mel1的红色荧光蛋白（RFP）融合体。然而，在营养细胞中，细胞壁无法保持RFP融合。尽管孢子壁阻止了蛋白质的扩散，但较小的分子（例如糖）很可能会穿过它。实际上，孢子可以比营养细胞含有更高的α-半乳糖苷酶活性来消化黑糖。当存在于孢子壁中时，该酶获得对环境压力的抗性，包括酶消化和高温。孢子壁的外层需要保留酶，但也会降低底物的可及性。但是，具有轻度孢子壁缺陷的突变体可以保留并稳定酶，同时仍然允许接近底物。除了Mel1，我们还显示了孢子可以保留转化酶。有趣的是，包封的转化酶对棉子糖的活性明显低于对蔗糖的活性。这表明通过封装可以改变基板的选择性。

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