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Chromosome-based strain engineering

    Chromosome-based strain engineering was feasible to alter the gene expression profile and further affect the overall metabolic pathway for chemical production. This strategy can prevent the cell burden from the plasmid-based regulation. Our studies focus on two aspects: one is regulating the protein expression through the level of RNA polymerase at different chromosome loci by site-specific phage attachment. The resulting protein expression significantly correlated with the gene dosage of RNA polymerase, paving the way for exploring more integration sites for gene integration in the future. The other is adaptive laboratory evolution (ALE) to improve strain performance. Compared to complex genomic engineering, adaptive-directed evolution was an alternative strategy to improve strain tolerance. In our study, we successfully perform the ALE to generate the evolved strain with better cell growth and chemical production.

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Related publications:

  • Wan-Wen Ting, I-Son Ng* (2022) Effective 5-aminolevulinic acid production via T7 RNA polymerase and RuBisCO equipped Escherichia coli W3110. Biotechnology Bioengineering 1–10. DOI: 10.1002/bit.28273

  • Wan-Wen Ting, Jie-Yao Yu, Yu-Chieh Lin, I-Son Ng* (2022) Enhanced recombinant carbonic anhydrase in T7RNAP-equipped Escherichia coli W3110 for carbon capture storage and utilization (CCSU). Bioresource Technology 363, 128010.

  • Sefli Sri Wahyu Effendi & I-Son Ng* (2022) Reprogramming T7RNA polymerase in Escherichia coli Nissle 1917 under specific lac operon for efficient p‑coumaric acid production. ACS Synthetic Biology 11, 3471-3481.

  • Ying-Chen Yi, Chengfeng Xue, I-Son Ng* (2021) Low carbon footprint production of high-end 5-aminolevulinic acid via integrative strain engineering and RuBisCo-equipped Escherichia coli. ACS Sustainable Chemistry & Engineering 9(46): 15623–15633

  • Shih-I Tan, I-Son Ng* (2021) Stepwise optimization of genetic RuBisCO-equipped Escherichia coli for low carbon-footprint protein and chemical production. Green Chemistry 23, 4800-4813.

  • Chengfeng Xue, Tzu-Hsuan Yu, I-Son Ng* (2021) Engineering pyridoxal kinase PdxY-integrated Escherichia coli strain and optimization for high-level 5-aminolevulinic acid production. Journal of the Taiwan Institute of Chemical Engineers 120: 49-58.

Biosensor & directed evolution 

Enzymes play an indispensable role as biocatalyst in the green process of biochemicals production. To strength the performance in harsh conditions, native enzymes are engineered with improvement of activity, selectivity, stability and tolerance. Directed evolution is one of the strategies to evolve the protein by random gene diversification and followed by the high-throughput screening. Our studies focus on the in vivo target-specific gene diversification by RNA polymerase or error-prone PCR (Ep-PCR) to create the gene mutation. For the enzyme screening, the metabolite-based biosensor was developed to screen from the fluorescence output. 

Related publications:

  • Shih-I Tan, I-Son Ng* (2021) CRISPRi-mediated NIMPLY logic gate for fine-tuning the whole-cell sensing toward simple urine glucose detection. ACS Synthetic Biology, 10(2), 412-421. doi.org/10.1021/acssynbio.1c00014

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