Abstract
BackgroundThe filamentous fungus Trichoderma reesei produces cellulase enzymes that are widely studied for lignocellulose bioconversion to biofuel. N,N-dimethylformamide (DMF) is a versatile organic solvent used in large quantities in industries.ResultsIn this study, we serendipitously found that biologically relevant concentrations of extracellular DMF-induced cellulase production in the T. reesei hyper-cellulolytic mutant Rut-C30 and wild-type strain QM6a. Next, by transcriptome analysis, we determined that plc-e encoding phospholipase C was activated by DMF and revealed that cytosolic Ca2+ plays a vital role in the response of T. reesei to DMF. Using EGTA (a putative extracellular Ca2+ chelator) and LaCl3 (a plasma membrane Ca2+ channel blocker), we demonstrated that DMF induced a cytosolic Ca2+ burst via extracellular Ca2+ and Ca2+ channels in T. reesei, and that the cytosolic Ca2+ burst induced by DMF-mediated overexpression of cellulase through calcium signaling. Deletion of crz1 confirmed that calcium signaling plays a dominant role in DMF-induced cellulase production. Additionally, 0.5–2% DMF increases the permeability of T. reesei mycelia for cellulase release. Simultaneous supplementation with 1% DMF and 10 mM Mn2+ to T. reesei Rut-C30 increased cellulase activity approximately fourfold compared to that without treatment and was also more than that observed in response to either treatment alone.ConclusionsOur results reveal that DMF-induced cellulase production via calcium signaling and permeabilization. Our results also provide insight into the role of calcium signaling in enzyme production for enhanced cellulase production and the development of novel inducers of cellulase.
Highlights
The filamentous fungus Trichoderma reesei produces cellulase enzymes that are widely studied for lignocellulose bioconversion to biofuel
Liu et al [17] demonstrated that calcium activates calmodulin to regulate ganoderic acid biosynthesis in Ganoderma lucidum, a well-known medicinal basidiomycete, and Chen et al [13] revealed that calcium signaling pathways participate in cellulase production in T. reesei
To further clarify the role of calcium signaling in DMF induction of cellulase overexpression, we investigated the effects of DMF induction on cellulase production in the crz1 deletion strain Δcrz1
Summary
The filamentous fungus Trichoderma reesei produces cellulase enzymes that are widely studied for lignocellulose bioconversion to biofuel. A wide diversity of induction signals, Chen et al Biotechnol Biofuels (2019) 12:36 including calcium signaling, are present during cellulase production and are able to regulate the expression of cellulase-encoding genes [12, 13]. Liu et al [17] demonstrated that calcium activates calmodulin to regulate ganoderic acid biosynthesis in Ganoderma lucidum, a well-known medicinal basidiomycete, and Chen et al [13] revealed that calcium signaling pathways participate in cellulase production in T. reesei. We demonstrated that Mn2+ induction of calcium signaling participates in regulating cellulase gene expression in T. reesei [18]
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