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Alleviating vacuolar transport improves cellulase production in Trichoderma reesei

  • Applied Genetics and Molecular Biotechnology
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Abstract

Increasing cellulase production in cellulolytic fungus Trichoderma reesei is of interest for biofuels and biorefineries. Previous studies indicated that secreted protein was occasionally accumulated in vacuoles; this phenomenon has also been reported in T. reesei. Therefore, alleviating vacuolar transport seems to be a promising strategy for improving cellulase production in T. reesei. Herein, we found that knockout of vps10, vps13, and vps21, among 11 vacuolar protein sorting factors, improved cellulase production in T. reesei. The filter paper activity in Δvps10, Δvps13, and Δvps21 increased by 1.28-, 2.45-, and 2.11-fold than that of the parent strain. Moreover, the β-glucosidase activity in Δvps13 and Δvps21 increased by 3.22- and 3.56-fold after 6 days of fermentation. Furthermore, we also found that the vacuolar trafficking towards vacuoles was partially impaired in three knockout mutants, and disruption of vps13 alleviated the autophagy process. These results indicated that alleviated transport and degradation towards vacuole in Δvps10, Δvps13, and Δvps21 might improve cellulase production. Of note, the expression of cellulase genes in Δvps13 and Δvps21 was dramatically increased in the late induction phase compared to the parent. These results suggested that Vps13 and Vps21 might influence the cellulase production at transcription level. And further transcriptome analysis indicated that increased cellulase gene expression might be attributed to the differential expression of sugar transporters. Our study unravels the effect of alleviating vacuolar transport through knockout vps10, vps13, and vps21 for efficient cellulase secretion, providing new clues for higher cellulase production in T. reesei.

Key points

• Disruption of vps10, vps13 or vps21 improves cellulase production

• Vacuolar transport is impaired in three vps KO mutants

• Deletion of vps13 or vps21 increases the transcript of cellulase genes in late stage

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Data availability

The authors declare that all data supporting the findings of this study are available within the paper and its supplementary information files. Moreover, the RNA-seq data have been deposited in the Gene Expression Omnibus database (GEO, http://www.ncbi.nlm.nih.gov/geo/) with accession number GSE187573. Other related figures and data are also available from the corresponding author upon request.

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Funding

This work is supported by the National Key Research and Development Program of China (2021YFC2100203); the National Natural Science Foundation of China (32072162); Postgraduate Research & Practice Innovation Program of Jiangsu Province (KYCX20_1819); National First-Class Discipline Program of Light Industry Technology and Engineering (LITE2018-09) and 111Project (111–2-06).

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  1. Lab of Brewing Microbiology and Applied Enzymology, School of Biotechnology and Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi, 214122, China

    Su Yan, Yan Xu & Xiao-Wei Yu

  2. State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, China

    Xiu-Mei Tao

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  1. Su Yan
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S.Y. and X.W.Y.: conceptualization. S.Y.: investigation; writing—original draft preparation. X.W.Y.: writing—review and editing. X.M.T.: methodology. Y. X. and X.W.Y.: supervision.

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Correspondence to Xiao-Wei Yu.

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Yan, S., Xu, Y., Tao, XM. et al. Alleviating vacuolar transport improves cellulase production in Trichoderma reesei. Appl Microbiol Biotechnol 107, 2483–2499 (2023). https://doi.org/10.1007/s00253-023-12478-4

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