Department of Biological Engineering, Inha University, Incheon 402-751, Korea.
J Microbiol Biotechnol. 2010 Feb;20(2):356-62.
Ceramide is important not only for the maintenance of the barrier function of the skin but also for the water-binding capacity of the stratum corneum. Although the exact role of ceramide in the human skin is not fully understood, ceramide has become a widely used ingredient in cosmetic and pharmaceutical industries. Compared to other microorganisms, yeast is more suitable for the production of ceramide because yeast grows fast and is non-toxic. However, production of ceramide from yeast has not been widely studied and most work in this area has been carried out using Saccharomyces cerevisiae. Regulating the genes that are involved in sphingolipd synthesis is necessary to increase ceramide production. In this study, we investigated the effect of the genes involved in the synthesis of ceramide, lcb1, lcb2, tsc10, lac1, lag1, and sur2 on ceramide production levels. The genes were cloned into pYES2 high copy number vectors. S. cerevisiae was cultivated on YPDG medium at 30 degrees Celsius. Ceramide was purified from the cell extracts by solvent extraction and the ceramide content was analyzed by HPLC using ELSD. The maximum production of ceramide (9.8 mg ceramide/g cell) was obtained when the tsc10 gene was amplified by the pYES2 vector. Real time PCR analysis showed that the increase in ceramide content was proportional to the increase in the tsc10 gene expression level, which was 4.56 times higher than that of the control strain.
神经酰胺不仅对皮肤的屏障功能至关重要，而且对角质层的保水能力也很重要。尽管神经酰胺在人体皮肤中的确切作用尚未完全了解，但它已成为化妆品和制药行业广泛使用的成分。与其他微生物相比，酵母更适合生产神经酰胺，因为酵母生长迅速且无毒。然而，从酵母中生产神经酰胺的研究并不广泛，该领域的大多数工作都是使用酿酒酵母（Saccharomyces cerevisiae）进行的。调节参与神经酰胺合成的基因对于提高神经酰胺的产量是必要的。在这项研究中，我们研究了参与神经酰胺合成的基因 lcb1、lcb2、tsc10、lac1、lag1 和 sur2 对神经酰胺生产水平的影响。这些基因被克隆到 pYES2 高拷贝数载体中。在 30°C 的 YPDG 培养基中培养酿酒酵母。通过溶剂萃取从细胞提取物中纯化神经酰胺，并通过 HPLC 结合蒸发光散射检测器（ELSD）分析神经酰胺的含量。当 tsc10 基因被 pYES2 载体扩增时，获得了最高的神经酰胺产量（9.8mg 神经酰胺/g 细胞）。实时 PCR 分析表明，神经酰胺含量的增加与 tsc10 基因表达水平的增加成正比，比对照菌株高 4.56 倍。
