Tags

Type your tag names separated by a space and hit enter

Enhancement of cellulase production in Trichoderma reesei RUT-C30 by comparative genomic screening.
Microb Cell Fact. 2019 May 10; 18(1):81.MC

Abstract

BACKGROUND

Cellulolytic enzymes produced by the filamentous fungus Trichoderma reesei are commonly used in biomass conversion. The high cost of cellulase is still a significant challenge to commercial biofuel production. Improving cellulase production in T. reesei for application in the cellulosic biorefinery setting is an urgent priority.

RESULTS

Trichoderma reesei hyper-cellulolytic mutant SS-II derived from the T. reesei NG14 strain exhibited faster growth rate and more efficient lignocellulosic biomass degradation than those of RUT-C30, another hyper-cellulolytic strain derived from NG14. To identify any genetic changes that occurred in SS-II, we sequenced its genome using Illumina MiSeq. In total, 184 single nucleotide polymorphisms and 40 insertions and deletions were identified. SS-II sequencing revealed 107 novel mutations and a full-length wild-type carbon catabolite repressor 1 gene (cre1). To combine the mutations of RUT-C30 and SS-II, the sequence of one confirmed beneficial mutation in RUT-C30, cre196, was introduced in SS-II to replace full-length cre1, forming the mutant SS-II-cre196. The total cellulase production of SS-II-cre196 was decreased owing to the limited growth of SS-II-cre196. In contrast, 57 genes mutated only in SS-II were selected and knocked out in RUT-C30. Of these, 31 were involved in T. reesei growth or cellulase production. Cellulase activity was significantly increased in five deletion strains compared with that in two starter strains, RUT-C30 and SS-II. Cellulase production of T. reesei Δ108642 and Δ56839 was significantly increased by 83.7% and 70.1%, respectively, compared with that of RUT-C30. The amount of glucose released from pretreated corn stover hydrolyzed by the crude enzyme from Δ108642 increased by 11.9%.

CONCLUSIONS

The positive attribute confirmed in one cellulase hyper-producing strain does not always work efficiently in another cellulase hyper-producing strain, owing to the differences in genetic background. Genome re-sequencing revealed novel mutations that might affect cellulase production and other pathways indirectly related to cellulase formation. Our strategy of combining the mutations of two strains successfully identified a number of interesting phenotypes associated with cellulase production. These findings will contribute to the creation of a gene library that can be used to investigate the involvement of various genes in the regulation of cellulase production.

Authors+Show Affiliations

State Key Lab of Bioreactor Engineering, East China University of Science and Technology, P.O.B. 311, 130 Meilong Road, Shanghai, 200237, China.State Key Lab of Bioreactor Engineering, East China University of Science and Technology, P.O.B. 311, 130 Meilong Road, Shanghai, 200237, China.State Key Lab of Bioreactor Engineering, East China University of Science and Technology, P.O.B. 311, 130 Meilong Road, Shanghai, 200237, China.State Key Lab of Bioreactor Engineering, East China University of Science and Technology, P.O.B. 311, 130 Meilong Road, Shanghai, 200237, China.State Key Lab of Bioreactor Engineering, East China University of Science and Technology, P.O.B. 311, 130 Meilong Road, Shanghai, 200237, China.Sunson Industry Group Co, Ltd, Beijing, China.State Key Lab of Bioreactor Engineering, East China University of Science and Technology, P.O.B. 311, 130 Meilong Road, Shanghai, 200237, China.State Key Lab of Bioreactor Engineering, East China University of Science and Technology, P.O.B. 311, 130 Meilong Road, Shanghai, 200237, China.State Key Lab of Bioreactor Engineering, East China University of Science and Technology, P.O.B. 311, 130 Meilong Road, Shanghai, 200237, China. wadexp@ecust.edu.cn.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31077201

Citation

Liu, Pei, et al. "Enhancement of Cellulase Production in Trichoderma Reesei RUT-C30 By Comparative Genomic Screening." Microbial Cell Factories, vol. 18, no. 1, 2019, p. 81.
Liu P, Lin A, Zhang G, et al. Enhancement of cellulase production in Trichoderma reesei RUT-C30 by comparative genomic screening. Microb Cell Fact. 2019;18(1):81.
Liu, P., Lin, A., Zhang, G., Zhang, J., Chen, Y., Shen, T., Zhao, J., Wei, D., & Wang, W. (2019). Enhancement of cellulase production in Trichoderma reesei RUT-C30 by comparative genomic screening. Microbial Cell Factories, 18(1), 81. https://doi.org/10.1186/s12934-019-1131-z
Liu P, et al. Enhancement of Cellulase Production in Trichoderma Reesei RUT-C30 By Comparative Genomic Screening. Microb Cell Fact. 2019 May 10;18(1):81. PubMed PMID: 31077201.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Enhancement of cellulase production in Trichoderma reesei RUT-C30 by comparative genomic screening. AU - Liu,Pei, AU - Lin,Aibo, AU - Zhang,Guoxiu, AU - Zhang,Jiajia, AU - Chen,Yumeng, AU - Shen,Tao, AU - Zhao,Jian, AU - Wei,Dongzhi, AU - Wang,Wei, Y1 - 2019/05/10/ PY - 2019/02/28/received PY - 2019/05/02/accepted PY - 2019/5/12/entrez PY - 2019/5/12/pubmed PY - 2019/7/23/medline KW - Alcohol dehydrogenase KW - CRE1 KW - Cellulase production KW - Genome sequencing KW - RUT-C30 KW - Trichoderma reesei KW - tre108642 KW - tre56839 SP - 81 EP - 81 JF - Microbial cell factories JO - Microb. Cell Fact. VL - 18 IS - 1 N2 - BACKGROUND: Cellulolytic enzymes produced by the filamentous fungus Trichoderma reesei are commonly used in biomass conversion. The high cost of cellulase is still a significant challenge to commercial biofuel production. Improving cellulase production in T. reesei for application in the cellulosic biorefinery setting is an urgent priority. RESULTS: Trichoderma reesei hyper-cellulolytic mutant SS-II derived from the T. reesei NG14 strain exhibited faster growth rate and more efficient lignocellulosic biomass degradation than those of RUT-C30, another hyper-cellulolytic strain derived from NG14. To identify any genetic changes that occurred in SS-II, we sequenced its genome using Illumina MiSeq. In total, 184 single nucleotide polymorphisms and 40 insertions and deletions were identified. SS-II sequencing revealed 107 novel mutations and a full-length wild-type carbon catabolite repressor 1 gene (cre1). To combine the mutations of RUT-C30 and SS-II, the sequence of one confirmed beneficial mutation in RUT-C30, cre196, was introduced in SS-II to replace full-length cre1, forming the mutant SS-II-cre196. The total cellulase production of SS-II-cre196 was decreased owing to the limited growth of SS-II-cre196. In contrast, 57 genes mutated only in SS-II were selected and knocked out in RUT-C30. Of these, 31 were involved in T. reesei growth or cellulase production. Cellulase activity was significantly increased in five deletion strains compared with that in two starter strains, RUT-C30 and SS-II. Cellulase production of T. reesei Δ108642 and Δ56839 was significantly increased by 83.7% and 70.1%, respectively, compared with that of RUT-C30. The amount of glucose released from pretreated corn stover hydrolyzed by the crude enzyme from Δ108642 increased by 11.9%. CONCLUSIONS: The positive attribute confirmed in one cellulase hyper-producing strain does not always work efficiently in another cellulase hyper-producing strain, owing to the differences in genetic background. Genome re-sequencing revealed novel mutations that might affect cellulase production and other pathways indirectly related to cellulase formation. Our strategy of combining the mutations of two strains successfully identified a number of interesting phenotypes associated with cellulase production. These findings will contribute to the creation of a gene library that can be used to investigate the involvement of various genes in the regulation of cellulase production. SN - 1475-2859 UR - https://www.unboundmedicine.com/medline/citation/31077201/Enhancement_of_cellulase_production_in_Trichoderma_reesei_RUT_C30_by_comparative_genomic_screening_ L2 - https://microbialcellfactories.biomedcentral.com/articles/10.1186/s12934-019-1131-z DB - PRIME DP - Unbound Medicine ER -