Abstract
The serine protease gene from a thermophilic fungus Thermoascus aurantiacus var. levisporus, was cloned, sequenced, and expressed in Pichia pastoris and the recombinant protein was characterized. The full-length cDNA of 2,592 bp contains an ORF of 1,482 bp encoding 494 amino acids. Sequence analysis of the deduced
amino acid sequence revealed high homology with subtilisin serine proteases. The putative enzyme contained catalytic domain with active sites formed by three residues of Asp183, His215, and Ser384. The molecular mass of the recombinant enzyme was estimated to be 59.1 kDa after overexpression in P. pastoris. The
activity of recombinant protein was 115.58 U/mg. The protease exhibited its maximal activity at 50°C and pH 8.0 and kept thermostable at 60°C, and retained 60% activity after 60 min at 70°C. The protease activity was found to be inhibited by PMSF, but not by DTT or EDTA. The enzyme has broad substrate specificity such as gelatin, casein and pure milk, and exhibiting highest activity towards casein.
Citations
Citations to this article as recorded by
- A Novel C-Terminal Small Tail Provides Thermostability of FeSOD Implying a New Mechanism of Protein Heat Resistance
Weina Lu, Zhuo Jiang, Qi Lin, Zhecheng Yang, Yanli Liu, Wenhui Bi, Zhengying You, Caiying Jiang, Qing Sheng, Zuoming Nie
The Protein Journal.2025;[Epub] CrossRef - Microbial proteases and their applications
Peng Song, Xue Zhang, Shuhua Wang, Wei Xu, Fei Wang, Rongzhao Fu, Feng Wei
Frontiers in Microbiology.2023;[Epub] CrossRef - Improving the catalytic activity of a detergent‐compatible serine protease by rational design
Xiao Wang, Xing Qin, Lige Tong, Jie Zheng, Tao Dong, Xiaolu Wang, Yuan Wang, Huoqing Huang, Bin Yao, Honglian Zhang, Huiying Luo
Microbial Biotechnology.2023; 16(5): 947. CrossRef - Identification of Exoenzymes Secreted by Entomopathogenic Fungus Beauveria pseudobassiana RGM 2184 and Their Effect on the Degradation of Cocoons and Pupae of Quarantine Pest Lobesia botrana
Matias Arias-Aravena, Fabiola Altimira, Daniela Gutiérrez, Jian Ling, Eduardo Tapia
Journal of Fungi.2022; 8(10): 1083. CrossRef - Heterologous expression and characterization of a novel subtilisin-like protease from a thermophilic Thermus thermophilus HB8
Guiqiu Xie, Zhengkang Shao, Li Zong, Xingxing Li, Dengli Cong, Rui Huo
International Journal of Biological Macromolecules.2019; 138: 528. CrossRef - Purification and characterization of cloned alkaline protease gene of Geobacillus stearothermophilus
Irfana Iqbal, Muhammad Nauman Aftab, Mohammed Afzal, Asad Ur‐Rehman, Saima Aftab, Asma Zafar, Zia Ud‐Din, Ateeque Rahman Khuharo, Jawad Iqbal, Ikram Ul‐Haq
Journal of Basic Microbiology.2015; 55(2): 160. CrossRef - Enhancing xylanase production in the thermophilic fungus Myceliophthora thermophila by homologous overexpression of Mtxyr1
Juan Wang, Yaning Wu, Yanfen Gong, Shaowen Yu, Gang Liu
Journal of Industrial Microbiology and Biotechnology.2015; 42(9): 1233. CrossRef - MS Analysis and Molecular Characterization of Botrytis cinerea Protease Prot-2. Use in Bioactive Peptides Production
Ferid Abidi, Nayssene Aissaoui, Jean-Charles Gaudin, Jean-Marc Chobert, Thomas Haertlé, Mohamed Nejib Marzouki
Applied Biochemistry and Biotechnology.2013; 170(2): 231. CrossRef - Characterization, cloning, and heterologous expression of a subtilisin-like serine protease gene VlPr1 from Verticillium lecanii
Gang Yu, Jin-Liang Liu, Li-Qin Xie, Xue-Liang Wang, Shi-Hong Zhang, Hong-Yu Pan
Journal of Microbiology.2012; 50(6): 939. CrossRef
MSK
Cite this Article
