Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Heterologous Expression and Characterization of a Thermostable α-L-Rhamnosidase from Thermoclostridium stercorarium subsp. thermolacticum DSM 2910 and Its Application in the Biotransformation of Rutin

  • Lin Ge (Department of Medical Science and Technology, Suzhou Chien-Shiung Institute of Technology) ;
  • Yingying Liu (Department of Medical Science and Technology, Suzhou Chien-Shiung Institute of Technology) ;
  • Fangming Zhou (Department of Medical Science and Technology, Suzhou Chien-Shiung Institute of Technology) ;
  • Lingling Zhan (Department of Medical Science and Technology, Suzhou Chien-Shiung Institute of Technology) ;
  • Linguo Zhao (Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, Nanjing Forestry University)
  • Received : 2023.05.28
  • Accepted : 2023.07.14
  • Published : 2023.11.28
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Abstract

An α-L-rhamnosidase gene from Thermoclostridium. stercorarium subsp. thermolacticum DSM 2910 (TstRhaA) was cloned and expressed. The maximum TstRhaA activity of the protein reached 25.2 U/ml, and the molecular mass was approximately 106.6 kDa. The protein was purified 8.0-fold by Ni-TED affinity with an overall recovery of 16.6% and a specific activity of 187.9 U/mg. TstRhaA activity was the highest at 65℃ and pH 6.5. In addition, it exhibited excellent thermal stability, better pH stability, good tolerance to low concentrations of organic reagents, and high catalytic activity for p-nitrophenyl-α-L-rhamnopyranoside (pNPR). Substrate specificity studies showed that TstRhaA exhibited a high specific activity for rutin. At 60℃, pH 6.5, and 0.3 U/ml enzyme dosage, 60 g/l rutin was converted to 45.55 g/l isoquercitrin within 150 min. The molar conversion rate of rutin and the yield of isoquercitrin were 99.8% and 12.22 g/l/h, respectively. The results suggested that TstRhaA could be used for mass production of isoquercitrin.

Keywords

Acknowledgement

This work was supported by the General Project of Natural Science Research of Jiangsu Province Higher Education Institutions (Grant No. 21KJB220015), and the Science and Technology Programme of Taicang (Grant No. TC2020JC17), "Two Funds" Project of Suzhou Chien-Shiung Institute of Technology (Grant No. JXLJ202208) as well as the start-up funds for doctoral research of Suzhou Chien-Shiung Institute of Technology.

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