The secretome of Pichia pastoris in fed-batch cultivations is largely independent of the carbon source but changes quantitatively over cultivation time.,Microbial Biotechnology

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The secretome of Pichia pastoris in fed-batch cultivations is largely independent of the carbon source but changes quantitatively over cultivation time.
Microbial Biotechnology ( IF 4.8 ) Pub Date : 2019-11-06 , DOI: 10.1111/1751-7915.13499
Jonas Burgard _{1,

2} , Clemens Grünwald-Gruber _{1,

3} , Friedrich Altmann _{1,

3} , Jürgen Zanghellini _{1,

2,

4} , Minoska Valli _{1,

2} , Diethard Mattanovich _{1,

2} , Brigitte Gasser _{1,

2}

Affiliation

The quantitative changes of the secretome of recombinant Pichia pastoris (Komagataella phaffii) CBS7435 over the time‐course of methanol‐ or glucose‐limited fed‐batch cultures were investigated by LC‐ESI‐MS/MS to define the carbon source‐specific secretomes under controlled bioreactor conditions. In both set‐ups, no indication for elevated cell lysis was found. The quantitative data revealed that intact and viable P. pastoris cells secrete only a low number of endogenous proteins (in total 51), even during high cell density cultivation. Interestingly, no marked differences in the functional composition of the P. pastoris secretome between methanol‐ and glucose‐grown cultures were observed with only few proteins being specifically affected by the carbon source. The ‘core secretome’ of 22 proteins present in all analysed carbon sources (glycerol, glucose and methanol) consists mainly of cell wall proteins. The quantitative analysis additionally revealed that most secretome proteins were already present after the batch phase, and depletion rather than accumulation occurred during the fed‐batch processes. Among the changes over cultivation time, the depletion of both the extracellularly detected chaperones and the only two identified proteases (Pep4 and Yps1‐1) during the methanol‐ or glucose‐feed phase appear as most prominent.

中文翻译：

补料分批培养中巴斯德毕赤酵母的分泌组在很大程度上与碳源无关，但随着培养时间的变化而发生定量变化。

通过LC-ESI-MS / MS研究了在甲醇或葡萄糖限制的分批分批培养物中随时间变化的重组巴斯德毕赤酵母CBS7435分泌组的定量变化，以定义在以下条件下碳源特定的分泌组受控的生物反应器条件。在这两种设置中，均未发现细胞裂解升高的迹象。定量数据表明，即使在高细胞密度培养过程中，完整且有活力的巴斯德毕赤酵母细胞仅分泌少量内源蛋白（总共51种）。有趣的是，巴斯德毕赤酵母的功能组成没有明显差异观察到甲醇和葡萄糖生长的培养物之间的分泌组，只有极少数的蛋白质受到碳源的特别影响。存在于所有分析碳源（甘油，葡萄糖和甲醇）中的22种蛋白质的“核心分泌组”主要由细胞壁蛋白质组成。定量分析还显示，在分批阶段之后已经存在大多数分泌组蛋白，在补料分批过程中发生的是消耗而不是积累。在培养时间的变化中，以甲醇或葡萄糖为原料的阶段中，细胞外检测到的伴侣分子和仅有的两种鉴定出的蛋白酶（Pep4和Yps1-1）的消耗最为明显。

更新日期：2019-11-06

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