PDF(Pubmed)
Abstract：
BACKGROUND: Human lysozyme (hLYZ) is a natural antibacterial protein with broad applications in food and pharmaceutical industries. Recombinant production of hLYZ in Komagataella phaffii (K. phaffii) has attracted considerable attention, but there are very limited strategies for its hyper-production in yeast.
RESULTS: Here through Atmospheric and Room Temperature Plasma (ARTP)-based mutagenesis and transcriptomic analysis, the expression of two genes MYO1 and IQG1 encoding the cytokinesis core proteins was identified downregulated along with higher hLYZ production. Deletion of either gene caused severe cytokinesis defects, but significantly enhanced hLYZ production. The highest hLYZ yield of 1,052,444 ± 23,667 U/mL bioactivity and 4.12 ± 0.11 g/L total protein concentration were obtained after high-density fed-batch fermentation in the Δmyo1 mutant, representing the best production of hLYZ in yeast. Furthermore, O-linked mannose glycans were characterized on this recombinant hLYZ.
CONCLUSIONS: Our work suggests that cytokinesis-based morphology engineering is an effective way to enhance the production of hLYZ in K. phaffii.
RESULTS: Here through Atmospheric and Room Temperature Plasma (ARTP)-based mutagenesis and transcriptomic analysis, the expression of two genes MYO1 and IQG1 encoding the cytokinesis core proteins was identified downregulated along with higher hLYZ production. Deletion of either gene caused severe cytokinesis defects, but significantly enhanced hLYZ production. The highest hLYZ yield of 1,052,444 ± 23,667 U/mL bioactivity and 4.12 ± 0.11 g/L total protein concentration were obtained after high-density fed-batch fermentation in the Δmyo1 mutant, representing the best production of hLYZ in yeast. Furthermore, O-linked mannose glycans were characterized on this recombinant hLYZ.
CONCLUSIONS: Our work suggests that cytokinesis-based morphology engineering is an effective way to enhance the production of hLYZ in K. phaffii.
摘要：
背景：人溶菌酶（hLYZ）是一种天然的抗菌蛋白，在食品和制药行业中具有广泛的应用。在Komagataellaphafii中重组生产hLYZ（K.phaffii)引起了相当大的关注，但是在酵母中它的高产有非常有限的策略。
结果：这里通过基于大气和室温等离子体（ARTP）的诱变和转录组学分析，编码胞质分裂核心蛋白的两个基因MYO1和IQG1的表达被鉴定为随着更高的hLYZ产生而下调。任一基因的缺失都会导致严重的胞质分裂缺陷，但显著提高了hLYZ的产量。在Δmyo1突变体中进行高密度补料分批发酵后，获得了最高的hLYZ产量1,052,444±23,667U/mL的生物活性和4.12±0.11g/L的总蛋白浓度，代表酵母中hLYZ的最佳产量。此外，在该重组hLYZ上表征O-连接的甘露糖聚糖。
结论：我们的工作表明，基于胞质分裂的形态学工程是增强K.phafii中hLYZ生产的有效方法。
结果：这里通过基于大气和室温等离子体（ARTP）的诱变和转录组学分析，编码胞质分裂核心蛋白的两个基因MYO1和IQG1的表达被鉴定为随着更高的hLYZ产生而下调。任一基因的缺失都会导致严重的胞质分裂缺陷，但显著提高了hLYZ的产量。在Δmyo1突变体中进行高密度补料分批发酵后，获得了最高的hLYZ产量1,052,444±23,667U/mL的生物活性和4.12±0.11g/L的总蛋白浓度，代表酵母中hLYZ的最佳产量。此外，在该重组hLYZ上表征O-连接的甘露糖聚糖。
结论：我们的工作表明，基于胞质分裂的形态学工程是增强K.phafii中hLYZ生产的有效方法。
