Increased Isoprene Production by the Recombinant Pantoea ananatis Strain due to the Balanced Amplification of Mevalonate Pathway Genes
- Authors: Katashkina J.I.1, Kazieva E.D.1, Tajima Y.2, Mashko S.V.1
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Affiliations:
- Ajinomoto-Genetika Research Institute (AGRI) Closed Joint-Stock Company
- Institute for Innovation, Ajinomoto Co., Inc., Kawasaki-ku
- Issue: Vol 55, No 9 (2019)
- Pages: 850-860
- Section: Producers, Biology, Selection, and Gene Engineering
- URL: https://journals.rcsi.science/0003-6838/article/view/153120
- DOI: https://doi.org/10.1134/S0003683819090023
- ID: 153120
Cite item
Abstract
The uncoordinated expression of a target biosynthetic pathway often results in low growth and productivity for a producing strain due to the accumulation of toxic metabolic intermediates. In this study, we report an increase in isoprene accumulation by an engineered strain of Pantoea ananatis that possesses foreign, artificial mevalonate-pathway genes. This was achieved due to the simultaneous, φ80-Int-dependent integration of a mixture of conditionally replicated plasmids carrying genes for the upper and lower mevalonate pathways and mevalonate kinase, followed by selection according to the isoprene accumulation of clones with optimal copy numbers of the integrative cassettes. A previously developed method, Dual In/Out, was modified to accelerate the construction of recipient strains containing several attB sites of the φ80 phage in their genomes.
About the authors
J. I. Katashkina
Ajinomoto-Genetika Research Institute (AGRI) Closed Joint-Stock Company
Email: ekaterina_kazieva@agri.ru
Russian Federation, Moscow, 117545
E. D. Kazieva
Ajinomoto-Genetika Research Institute (AGRI) Closed Joint-Stock Company
Author for correspondence.
Email: ekaterina_kazieva@agri.ru
Russian Federation, Moscow, 117545
Y. Tajima
Institute for Innovation, Ajinomoto Co., Inc., Kawasaki-ku
Email: ekaterina_kazieva@agri.ru
Japan, Kawasaki, 210-8681
S. V. Mashko
Ajinomoto-Genetika Research Institute (AGRI) Closed Joint-Stock Company
Email: ekaterina_kazieva@agri.ru
Russian Federation, Moscow, 117545
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