背景:人溶菌酶(hLYZ),一种新兴的抗菌剂,在食品和制药行业有广泛的应用。然而,hLYZ的来源特别有限。
结果:为了在毕赤酵母中实现人溶菌酶的高效表达和分泌,多种策略,包括G418硫酸盐筛查,信号序列优化,应用液泡分选受体VPS10破坏和伴侣/转录因子共表达。摇瓶中细胞外hLYZ的最大酶活性为81,600±5230U·mL-1,大约是原始菌株的5倍。为了进一步降低成本,开发了最佳培养基RDMY,在5L发酵罐中最高hLYZ活性达到352,000±16696.5U·mL-1。
结论:这项研究为巴斯德毕赤酵母中的hLYZ生产提供了一种非常有用且具有成本效益的方法,也可以应用于其他重组蛋白的生产。本文受版权保护。保留所有权利。
BACKGROUND: Human lysozyme (hLYZ), an emerging antibacterial agent, has extensive application in the food and pharmaceutical industries. However, the source of hLYZ is particularly limited.
RESULTS: To achieve highly efficient expression and secretion of hLYZ in Pichia pastoris, multiple strategies including G418 sulfate screening, signal sequence optimization, vacuolar sorting receptor VPS10 disruption, and chaperones/transcription factors co-expression were applied. The maximal enzyme activity of extracellular hLYZ in a shaking flask was 81,600 ± 5230 U mL-1 , which was about five times of original strain. To further reduce the cost, the optimal medium RDMY was developed and the highest hLYZ activity reached 352,000 ± 16,696.5 U mL-1 in a 5 L fermenter.
CONCLUSIONS: This research provides a very useful and cost-effective approach for the hLYZ production in P. pastoris and can also be applied to the production of other recombinant proteins.