Lipomax is a commercialized foldase-dependent Pseudomonas lipase that was previously expressed only in Pseudomonas strains. Here, using Pichia pastoris as the host, we report a new co-expression method that leads to the successful production of Lipomax. The active Lipomax is extracellularly co-expressed with its cognate foldase (LIM); and the purified enzyme mix has the optimum pH at pH 8.0 and an optimal temperature around 40°C. N-glycosylation was observed for Pichia produced Lipomax, and its reduction was shown to increase the lipolytic activity. With different p-nitrophenyl esters as the substrates, the substrate profiling analyses further indicate that Lipomax prefers esters with middle-long chain fatty acids, showing the highest specific activity to p-nitrophenyl caprylate (C8). The extracellular co-expression of Lipomax and LIM in Pichia will not only increase our ability to investigate additional eukaryotic hosts for lipase expression, but also be of considerable value in analyzing other foldase-dependent lipases.

Lipomax是一种商业化的依赖折叠酶的假单胞菌脂肪酶，以前仅在假单胞菌菌株中表达。在这里，我们以巴斯德毕赤酵母为宿主，报道了一种新的共表达方法，该方法可成功生产Lipomax。活性Lipomax与它的同源折叠酶（LIM）在细胞外共表达；纯化的酶混合物在pH 8.0时具有最佳pH，在40°C时具有最佳温度。观察到毕赤酵母产生的Lipomax的N-糖基化，并且其减少显示出增加的脂解活性。与不同的p-以硝基苯基酯为底物，底物谱分析进一步表明，Lipomax更偏爱具有中长链脂肪酸的酯，与对硝基苯基辛酸酯（C8）相比具有最高的比活性。Lipomax和LIM在毕赤酵母中的细胞外共表达，不仅会增强我们研究脂肪酶表达的其他真核宿主的能力，而且在分析其他依赖折叠酶的脂肪酶方面也具有重要价值。