Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Insertion Mutation in HMG-CoA Lyase Increases the Production Yield of MPA through Agrobacterium tumefaciens-Mediated Transformation

  • Dong, Yuguo (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology) ;
  • Zhang, Jian (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology) ;
  • Xu, Rui (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology) ;
  • Lv, Xinxin (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology) ;
  • Wang, Lihua (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology) ;
  • Sun, Aiyou (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology) ;
  • Wei, Dongzhi (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology)
  • Received : 2016.03.18
  • Accepted : 2016.07.29
  • Published : 2016.11.28
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Abstract

Mycophenolic acid (MPA) is an antibiotic produced by Penicillium brevicompactum. MPA has antifungal, antineoplastic, and immunosuppressive functions, among others. ${\beta}-Hydroxy-{\beta}-methylglutaryl-CoA$ (HMG-CoA) lyase is a key enzyme in the bypass metabolic pathway. The inhibitory activity of HMG-CoA lyase increases the MPA biosynthetic flux by reducing the generation of by-products. In this study, we cloned the P. brevicompactum HMG-CoA lyase gene using the thermal asymmetric interlaced polymerase chain reaction and gene walking technology. Agrobacterium tumefaciens-mediated transformation (ATMT) was used to insert a mutated HMG-CoA lyase gene into P. brevicompactum. Successful insertion of the HMG-CoA lyase gene was confirmed by hygromycin screening, PCR, Southern blot analysis, and enzyme content assay. The maximum MPA production by transformants was 2.94 g/l. This was 71% higher than wild-type ATCC 16024. Our results demonstrate that ATMT may be an alternative practical genetic tool for directional transformation of P. brevicompactum.

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