Expression and Renaturation Analysis of Adenylosuccinate Synthetase from Cordyceps cicadae

Tianjin Key Laboratory of Food Science and Biotechnology, College of Biotechnology and Food Science, Tianjin University of Commerce, Tianjin, China

^* Corresponding author: ruanhaihua@tjcu.edu.cn

Abstract

In this study, the objective was to obtain adenylosuccinate synthase (ADSS) in the adenosine biosynthesis pathway of Cordyceps cicadae. The coding sequence (CDS) information of the protein was obtained from transcriptome sequencing. Bioinformatics analysis were employed to predict and analyze the physicochemical properties, hydrophobicity, signal peptide, and advanced structure of ADSS. The ADSS coding sequence was chemically synthesized and a prokaryotic expression vector containing N-terminal His tag (pET28a-ADSS) was constructed. Induction of ADSS expression occurred with 0.1 mmol/L IPTG at 37°C for 2 hours. SDS-PAGE was utilized to detect soluble protein expression levels. Although ADSS primarily expressed as inclusion bodies, soluble proteins were successfully obtained after denaturation and renaturation.