Li Ni, Zeng Yi, Chen Yumeng, Shen Yaling, Wang Wei
The State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, China.
Bioresour Bioprocess. 2022 Sep 6;9(1):96. doi: 10.1186/s40643-022-00587-3.
Trichoderma reesei RUT-C30 is a well-known high-yielding cellulase-producing fungal strain that converts lignocellulose into cellulosic sugar for resource regeneration. Calcium is a ubiquitous secondary messenger that regulates growth and cellulase production in T. reesei. We serendipitously found that adding Sr to the medium significantly increased cellulase activity in the T. reesei RUT-C30 strain and upregulated the expression of cellulase-related genes. Further studies showed that Sr supplementation increased the cytosolic calcium concentration and activated the calcium-responsive signal transduction pathway of Ca-calcineurin-responsive zinc finger transcription factor 1 (CRZ1). Using the plasma membrane Ca channel blocker, LaCl, we demonstrated that Sr induces cellulase production via the calcium signaling pathway. Supplementation with the corresponding concentrations of Sr also inhibited colony growth. Sr supplementation led to an increase in intracellular reactive oxygen species (ROS) and upregulated the transcriptional levels of intracellular superoxide dismutase (sod1) and catalase (cat1). We further demonstrated that ROS content was detrimental to cellulase production, which was alleviated by the ROS scavenger N-acetyl cysteine (NAC). This study demonstrated for the first time that Sr supplementation stimulates cellulase production and upregulates cellulase genes via the calcium signaling transduction pathway. Sr leads to an increase in intracellular ROS, which is detrimental to cellulase production and can be alleviated by the ROS scavenger NAC. Our results provide insights into the mechanistic study of cellulase synthesis and the discovery of novel inducers of cellulase.
里氏木霉RUT-C30是一种著名的高产纤维素酶产生菌株,可将木质纤维素转化为纤维素糖以进行资源再生。钙是一种普遍存在的第二信使,可调节里氏木霉的生长和纤维素酶产生。我们意外地发现,向培养基中添加Sr可显著提高里氏木霉RUT-C30菌株的纤维素酶活性,并上调纤维素酶相关基因的表达。进一步研究表明,添加Sr可增加胞质钙浓度,并激活钙调磷酸酶反应性锌指转录因子1(CRZ1)的钙响应信号转导途径。使用质膜钙通道阻滞剂LaCl,我们证明Sr通过钙信号通路诱导纤维素酶产生。补充相应浓度的Sr也会抑制菌落生长。添加Sr导致细胞内活性氧(ROS)增加,并上调细胞内超氧化物歧化酶(sod1)和过氧化氢酶(cat1)的转录水平。我们进一步证明,ROS含量对纤维素酶产生有害,而ROS清除剂N-乙酰半胱氨酸(NAC)可缓解这种有害作用。本研究首次证明,添加Sr通过钙信号转导途径刺激纤维素酶产生并上调纤维素酶基因。Sr导致细胞内ROS增加,这对纤维素酶产生有害,而ROS清除剂NAC可缓解这种有害作用。我们的结果为纤维素酶合成的机制研究和新型纤维素酶诱导剂的发现提供了见解。