The whole-cell (WC) bioconversion, considered as in vitro biotransformation, has been proposed to be a recyclable, tunable, and scalable system without concerns of cell proliferation, complex metabolic regulation, and byproduct formation, thus reducing cost and conversion time. Our group aimed in screening the key enzymes from enzyme databases that had highly active and stable properties, followed by tailoring and expressing ACN and CAD in E. coli BL21(DE3). Finally, the engineered Escherichia coli expressing dual genes and cultured in glycerol-included medium reached the highest IA titer of 67 g/L and productivity of 8.375 g/L/h, which demonstrates as a promising renewable process. All findings including using genetic design of cascade biocatalyst, modifying the culture medium, and optimizing the bioconversion parameters through Taguchi L9 array provides essential insights into designing the cascade enzymes for cost-effective and high-level IA titer.​

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Publication:

Chuan-Chieh Hsiang, Priskila Adjani Diankristanti, Shih-I Tan, Yi-Chia Ke, Yeong-Chang Chen, Sefli Sri Wahyu Effendi, I-Son Ng* (2022) Tailoring key enzymes for renewable and high-level itaconic acid production using genetic Escherichia coli via whole-cell bioconversion. Enzyme and Microbial Technology, 160, 110087.