Author:
Li Jinyang,Wang Yidi,Yang Kun,Wang Xiaolu,Wang Yuan,Zhang Honglian,Huang Huoqing,Su Xiaoyun,Yao Bin,Luo Huiying,Qin Xing
Abstract
Abstract
Background
Thermophilic fungus Myceliophthora thermophila has been widely used in industrial applications due to its ability to produce various enzymes. However, the lack of an efficient protein expression system has limited its biotechnological applications.
Results
In this study, using a laccase gene reporting system, we developed an efficient protein expression system in M. thermophila through the selection of strong constitutive promoters, 5’UTRs and signal peptides. The expression of the laccase was confirmed by enzyme activity assays. The results showed that the Mtpdc promoter (Ppdc) was able to drive high-level expression of the target protein in M. thermophila. Manipulation of the 5’UTR also has significant effects on protein expression and secretion. The best 5’UTR (NCA-7d) was identified. The transformant containing the laccase gene under the Mtpdc promoter, NCA-7d 5’UTR and its own signal peptide with the highest laccase activity (1708 U/L) was obtained. In addition, the expression system was stable and could be used for the production of various proteins, including homologous proteins like MtCbh-1, MtGh5-1, MtLPMO9B, and MtEpl1, as well as a glucoamylase from Trichoderma reesei.
Conclusions
An efficient protein expression system was established in M. thermophila for the production of various proteins. This study provides a valuable tool for protein production in M. thermophila and expands its potential for biotechnological applications.
Funder
China Agriculture Research System of Ministry of Finance (MOF) and Ministry of Agriculture and Rural Affairs
National Key Research and Development Program of China
Agricultural Science and Technology Innovation Program
Publisher
Springer Science and Business Media LLC
Subject
Applied Microbiology and Biotechnology,Bioengineering,Biotechnology