The filamentous fungus Trichoderma reesei is extensively utilized in the realm of recombinant protein expression owing to its well-established protein production systems. However, the potential for efficient and convenient protein production in T. reesei has not been fully harnessed. To further increase the production of recombinant lipase Candida antarctica lipase B (CalB), we overexpressed seven transcription activators and found that overexpression of the calcineurin CRZ1 could significantly enhance CalB production by 2.08-fold, and then through employing a synthetic strong promoter Pcbh7IR and knocking out ace1, a competitor of crz1, CalB production was further increased 4.2-fold. Furthermore, we found that increasing the Ca²⁺ concentration enabled the enhancement of CalB production in the CRZ1-overexpressing strains. Additionally, transcription analysis revealed a significant upregulation of calcium-related pathway genes and ER chaperone genes in T. reesei, revealing that overexpression of CRZ1 can increase the production of the recombinant protein by activation of the calcium signaling pathway. In this study, by systematically manipulating the recombinant protein expression module and its regulatory transcription activators, we found a positive effect of crz1-mediated calcium channels in boosting the expression of the recombinant protein.

中文翻译：

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CRZ1 在里氏木霉中的组成型过表达揭示了其增强重组脂肪酶产生的能力


丝状真菌 Trichoderma reesei 由于其完善的蛋白质生产系统而被广泛用于重组蛋白表达领域。然而，里氏锥虫高效便捷的蛋白质生产潜力尚未得到充分利用。为了进一步增加重组脂肪酶南极念珠菌脂肪酶 B （CalB） 的产生，我们过表达了 7 种转录激活因子，发现钙调磷酸酶 CRZ1 的过表达可以显着提高 CalB 的产生 2.08 倍，然后通过使用合成的强启动子 Pcbh7IR 并敲除 crz1 的竞争对手 ace1，CalB 的产生进一步增加了 4.2 倍。此外，我们发现增加 Ca^{2 +} 浓度能够增强 CRZ1 过表达菌株中 CalB 的产生。此外，转录分析显示 T. reesei 中钙相关通路基因和 ER 伴侣基因显着上调，揭示了 CRZ1 的过表达可以通过激活钙信号通路来增加重组蛋白的产生。在这项研究中，通过系统地操纵重组蛋白表达模块及其调节转录激活剂，我们发现 crz1 介导的钙通道在促进重组蛋白表达方面具有积极作用。