Abstract
Purpose
This study aimed to reveal the roles of the protein kinase A catalytic subunit 1 (pkac1) and carbon catabolite repressor cre1 genes in cellulase production by Trichoderma reesei wild-type strain QM6a. Our strategy might be useful to construct a high-yielding cellulase strain for its wide application.
Methods
This paper describes cellulase activity, plate conidiation, and yellow pigment synthesis assays of QM6a with the disruption of pkac1 and cre1.
Results
Deletion of pkac1 (Δpkac1) had no effect on cellulase production or transcript levels of major cellulase genes in the presence of cellulose. Disruption of cre1 (Δcre1) resulted in a remarkable increase in cellulase production and expression of the four major cellulase genes. Double disruption of pkac1 and cre1 significantly improved enzyme activity and protein production. The double disruption also resulted in a significant reduction in yellow pigment production and abrogated conidial production.
Conclusion
Double deletion of pkac1 and cre1 led to increased hydrolytic enzyme production in T. reesei using cellulose as a carbon source.
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Abbreviations
- CCR:
-
Representing carbon catabolite repressor
- CMCase:
-
Representing endo-β-glucanase activity
- FPase:
-
Filter paper hydrolase, representing total extracellular cellulase activity
- PKA:
-
Representing protein kinase A
- pNPCase:
-
Representing exo-β-glucanase activity
- RT-qPCR:
-
Representing real-time quantitative PCR
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Funding
This work was supported by the Natural Science Foundation of Shanghai, China (No. 22ZR1417600), the National Natural Science Foundation of China (32000050), and Shanghai Agriculture Applied Technology Development Program, China (Grant No. 2021-02-08-00-12-F00758).
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WW conceived and directed the study and reviewed the manuscript. NL carried out experiments and measurements, analyzed the data and wrote the manuscript. YC interpreted experimental data and supported the research funding. YS reviewed the manuscript. All authors have read and approved the final manuscript.
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Li, N., Chen, Y., Shen, Y. et al. Roles of PKAc1 and CRE1 in cellulose degradation, conidiation, and yellow pigment synthesis in Trichoderma reesei QM6a. Biotechnol Lett 44, 1465–1475 (2022). https://doi.org/10.1007/s10529-022-03312-4
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DOI: https://doi.org/10.1007/s10529-022-03312-4