Insight into the thermostability of thermophilic L-asparaginase and non-thermophilic L-asparaginase II through bioinformatics and structural analysis
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Thermostability plays an important role in the application of L-asparaginase in the pharmaceutical and food industries. Understanding the key residues and structures that influence thermostability in L-asparaginase is necessary to obtain suitable L-asparaginase candidates. In this study, special residues and structures that altered thermostability in thermophilic L-asparaginase and non-thermophilic L-asparaginase II were identified. Interchanging these special residues and structures of L-asparaginases from the four strains, that is, Pyrococcus yayanosii CH1 (PYA), Thermococcus gammatolerans (TGA), Bacillus subtilis (BSA II), and Escherichia coli (ECA II), revealed the 51st and 298th residues of PYA (corresponding to 57th, 305th residues of ECA II) as the key residues responsible for thermal stability of thermophilic L-asparaginase and non-thermophilic L-asparaginase II. Moreover, the C terminal tightness, loop rigidity, and low surface charge around activity sites were of great significance to the thermostability of L-asparaginase. This study therefore revealed the crucial amino acid residues and structures responsible for the difference in thermostability of the thermophilic and non-thermophilic L-asparaginase and provides a reference for engineering thermostability in L-asparaginase II.
KeywordsL-Asparaginase Thermostability Mutagenesis Bioinformatics Structure analysis
This work was supported by the National Natural Science Foundation of China (31500065, 31870066), the National Key Research and Development Program of China (2018YFA090039), the Fundamental Research Funds for the Central Universities (JUSRP51708A), the Program of the Key Laboratory of Industrial Biotechnology, Ministry of Education, China (KLIB-KF201703), the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, the 111 Project (111-2-06), the national first-class discipline program of Light Industry Technology and Engineering (LITE2018-06), the Jiangsu province “Collaborative Innovation Center for Modern Industrial Fermentation” industry development program, the Key Research and Development Program of Ningxia Hui Autonomous Region (2017BY069), the Science and Technology Innovation Team Foundation of Ningxia Hui Autonomous Region (KJT2017001), and the Ningxia Key Laboratory for Food Microbial-Applications Technology and Safety Control.
Compliance with ethical standards
The authors declare that they have no competing interests.
This article does not contain any studies with human participants or animals performed by any of the authors.
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