通过减压和甲醇诱导优化改善Pichia pastoris发酵中的植物酶生产
Carlos H Luna-Flores1, Yilun Weng2, Alexander Wang3
1Faculty of Science, Queensland University of Technology (QUT), Brisbane, Queensland, Australia.
Biotechnology and bioengineering
|July 25, 2023
概括
这项研究使用两阶段发酵过程优化了Pichia pastoris中的植物酶生产. 将葡萄糖用于生物质和甲醇用于诱导的结合显著提高了植物酶产量,达到12.65 MU/L.
科学领域:
- 生物技术是生物技术.
- 微生物工程 微生物工程
- 蛋白质生产 蛋白质生产
背景情况:
- 牧羊 (Komagataella phaffii) 是一种以高效的异质蛋白质分泌而闻名的甲基转化酵母.
- 之前在P. pastoris中进行的植物酶生产研究仅限于摇瓶,不反映工业规模的条件.
研究的目的:
- 使用仪器发酵器优化P. pastoris中的植物酶表达和生产.
- 开发和验证一种动力模型,用于在连续和料批量条件下预测植物酶生产.
主要方法:
- 在各种稀释系数 (0.5,1,1.5d-1) 的连续培养中研究了植物酶的产生.
- 开发了一个动力模型,使用发酵器数据来告知料批次培养设计.
- 采用了两阶段的发酵策略:葡萄糖用于生物质和去压缩,其次是甲醇用于诱导.
主要成果:
- 通过动态建模,在0.041h-1的特定生长率下确定了最佳的植物酶生产率.
- 与单独的葡萄糖相比,通过甲醇诱导实现了植物酶产量的3.5倍增加.
- 达到12.65 MU/L的最终植物酶活性,比瓶式发酵提高了36倍.
结论:
- 两阶段的发酵过程 (生物质的葡萄糖,诱导的甲醇) 对于工程P. pastoris的高植物酶产量至关重要.
- 开发的运动模型准确地预测了植物酶的产生,有助于生物过程的优化.
- 这项研究提出了一种可概括的方法,用于优化P. pastoris.中的蛋白质生产.
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