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Kinetic Separation in Escherichia coli by Developing Non-native Enzyme for Itaconate Production
Daeyeol YE,Myung Hyun NOH,Johyun MOON,Jeong Wook LEE,Jae-Seong YANG,Gyoo Yeol JUNG 한국생물공학회 2021 한국생물공학회 학술대회 Vol.2021 No.10
Microbial production of itaconate has been mainly conducted by Aspergillus terreus. Recently, its production in Escherichia coli has been researched due to the complicated fermentation process and shortage of genetic engineering tools of A. terreus. Unilke A. terreus, spatial separation of metabolic flux for the itaconate production has been hardly feasible in E. coli, which leads to the low level of its precursor, cis-aconitate. In this study, directed evolution of protein known to have substrate promiscuity to citrate was conducted in E. coli to enhance the cis-aconitate availability with the help of itaconate responsive screening system. Thereafter, the developed mutant showed significantly enhanced catalytic efficiency to citrate compared to the wild-type enzyme and consequently, its use enabled significantly increased itaconate production compared to the parental strain, indicating the successful enhancement of cis-aconitate supply by kinetic separation.