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High-level expression of highly active and thermostable trehalase from Myceliophthora thermophila in Aspergillus niger by using the CRISPR/Cas9 tool and its application in ethanol fermentation.
J Ind Microbiol Biotechnol. 2020 Jan; 47(1):133-144.JI

Abstract

Trehalase catalyzes the hydrolysis of the non-reducing disaccharide trehalose. The highly active trehalase MthT from Myceliophthora thermophila was screened from the trehalase genes of six species of filamentous fungi. An ingenious multi-copy knock-in expression strategy mediated by the CRISPR/Cas9 tool and medium optimization were used to improve MthT production in Aspergillus niger, up to 1698.83 U/mL. The protein background was dramatically abated due to insertion. The recombinant MthT showed optimal activity at pH 5.5 and 60 °C, and exhibited prominent thermal stability between 50 and 60 °C under acid conditions (pH 4.5-6.5). The ethanol conversion rate (ethanol yield/total glucose) was significantly improved by addition of MthT (51.88%) compared with MthT absence (34.38%), using 30% starch saccharification liquid. The results of this study provided an effective strategy, established a convenient platform for heterologous expression in A. niger and showed a potential strategy to decrease production costs in industrial ethanol production.

Authors+Show Affiliations

Room 315 Building B6, School of Biology and Biological Engineering, South China University of Technology, Guangzhou Higher Education Mega Center, Panyu town, Guangzhou, 510006, Guangdong, China.Room 315 Building B6, School of Biology and Biological Engineering, South China University of Technology, Guangzhou Higher Education Mega Center, Panyu town, Guangzhou, 510006, Guangdong, China.Room 315 Building B6, School of Biology and Biological Engineering, South China University of Technology, Guangzhou Higher Education Mega Center, Panyu town, Guangzhou, 510006, Guangdong, China.Room 315 Building B6, School of Biology and Biological Engineering, South China University of Technology, Guangzhou Higher Education Mega Center, Panyu town, Guangzhou, 510006, Guangdong, China.Room 315 Building B6, School of Biology and Biological Engineering, South China University of Technology, Guangzhou Higher Education Mega Center, Panyu town, Guangzhou, 510006, Guangdong, China. Guangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, Guangzhou, 510006, China.Room 315 Building B6, School of Biology and Biological Engineering, South China University of Technology, Guangzhou Higher Education Mega Center, Panyu town, Guangzhou, 510006, Guangdong, China. btlipan@scut.edu.cn. Guangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, Guangzhou, 510006, China. btlipan@scut.edu.cn.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31786675

Citation

Dong, Liangbo, et al. "High-level Expression of Highly Active and Thermostable Trehalase From Myceliophthora Thermophila in Aspergillus Niger By Using the CRISPR/Cas9 Tool and Its Application in Ethanol Fermentation." Journal of Industrial Microbiology & Biotechnology, vol. 47, no. 1, 2020, pp. 133-144.
Dong L, Lin X, Yu D, et al. High-level expression of highly active and thermostable trehalase from Myceliophthora thermophila in Aspergillus niger by using the CRISPR/Cas9 tool and its application in ethanol fermentation. J Ind Microbiol Biotechnol. 2020;47(1):133-144.
Dong, L., Lin, X., Yu, D., Huang, L., Wang, B., & Pan, L. (2020). High-level expression of highly active and thermostable trehalase from Myceliophthora thermophila in Aspergillus niger by using the CRISPR/Cas9 tool and its application in ethanol fermentation. Journal of Industrial Microbiology & Biotechnology, 47(1), 133-144. https://doi.org/10.1007/s10295-019-02252-9
Dong L, et al. High-level Expression of Highly Active and Thermostable Trehalase From Myceliophthora Thermophila in Aspergillus Niger By Using the CRISPR/Cas9 Tool and Its Application in Ethanol Fermentation. J Ind Microbiol Biotechnol. 2020;47(1):133-144. PubMed PMID: 31786675.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - High-level expression of highly active and thermostable trehalase from Myceliophthora thermophila in Aspergillus niger by using the CRISPR/Cas9 tool and its application in ethanol fermentation. AU - Dong,Liangbo, AU - Lin,Xiaotong, AU - Yu,Dou, AU - Huang,Lianggang, AU - Wang,Bin, AU - Pan,Li, Y1 - 2019/11/30/ PY - 2019/07/12/received PY - 2019/11/13/accepted PY - 2019/12/2/pubmed PY - 2020/5/15/medline PY - 2019/12/2/entrez KW - Aspergillus niger KW - CRISPR/Cas9 KW - Ethanol fermentation KW - Multi-copy KW - Trehalase SP - 133 EP - 144 JF - Journal of industrial microbiology & biotechnology JO - J. Ind. Microbiol. Biotechnol. VL - 47 IS - 1 N2 - Trehalase catalyzes the hydrolysis of the non-reducing disaccharide trehalose. The highly active trehalase MthT from Myceliophthora thermophila was screened from the trehalase genes of six species of filamentous fungi. An ingenious multi-copy knock-in expression strategy mediated by the CRISPR/Cas9 tool and medium optimization were used to improve MthT production in Aspergillus niger, up to 1698.83 U/mL. The protein background was dramatically abated due to insertion. The recombinant MthT showed optimal activity at pH 5.5 and 60 °C, and exhibited prominent thermal stability between 50 and 60 °C under acid conditions (pH 4.5-6.5). The ethanol conversion rate (ethanol yield/total glucose) was significantly improved by addition of MthT (51.88%) compared with MthT absence (34.38%), using 30% starch saccharification liquid. The results of this study provided an effective strategy, established a convenient platform for heterologous expression in A. niger and showed a potential strategy to decrease production costs in industrial ethanol production. SN - 1476-5535 UR - https://www.unboundmedicine.com/medline/citation/31786675/High-level_expression_of_highly_active_and_thermostable_trehalase_from_Myceliophthora_thermophila_in_Aspergillus_niger_by_using_the_CRISPR/Cas9_tool_and_its_application_in_ethanol_fermentation L2 - http://dx.doi.org/10.1007/s10295-019-02252-9 DB - PRIME DP - Unbound Medicine ER -