Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Luo-Yu Road #1037, Wuhan, 430074, China.
Appl Biochem Biotechnol. 2020 Jan;190(1):218-231. doi: 10.1007/s12010-019-03077-z. Epub 2019 Jul 22.
Yeast surface display has emerged as a viable approach for self-immobilization enzyme as whole-cell catalysts. Herein, we displayed Candida rugosa lipase 1 (CRL LIP1) on the cell wall of Pichia pastoris for docosahexaenoic acid (DHA) enrichment in algae oil. After a 96-h culture, the displayed CRL LIP1 achieved the highest activity (380 ± 2.8 U/g) for hydrolyzing olive oil under optimal pH (7.5) and temperature (45 °C) conditions. Additionally, we improved the thermal stability of displayed LIP1, enabling retention of 50% of its initial bioactivity following 6 h of incubation at 45 °C. Furthermore, the content of DHA enhanced from 40.61% in original algae oil to 50.44% in glyceride, resulting in a 1.24-fold increase in yield. The displayed CRL LIP1 exhibited an improved thermal stability and a high degree of bioactivity toward its native macromolecule substrates algae oil and olive oil, thereby expanding its potential for industrial applications in fields of food and pharmaceutical. These results suggested that surface display provides an effective strategy for simultaneous convenient expression and target protein immobilization.
酵母表面展示已成为将酶作为全细胞催化剂进行自我固定的一种可行方法。在此，我们将 Candida rugosa 脂肪酶 1（CRL LIP1）展示在毕赤酵母细胞壁上，用于藻类油中二羟基二十碳六烯酸（DHA）的富集。经过 96 小时的培养，展示的 CRL LIP1 在最佳 pH（7.5）和温度（45°C）条件下对橄榄油的水解活性最高（380±2.8 U/g）。此外，我们提高了展示的 LIP1 的热稳定性，使其在 45°C 孵育 6 小时后仍能保持其初始生物活性的 50%。此外，DHA 的含量从原始藻类油中的 40.61%提高到甘油酯中的 50.44%，产率提高了 1.24 倍。展示的 CRL LIP1 表现出改善的热稳定性和对其天然大分子底物藻类油和橄榄油的高度生物活性，从而扩大了其在食品和制药等领域的工业应用潜力。这些结果表明，表面展示为目标蛋白的同时方便表达和固定化提供了一种有效的策略。
