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Biotechnol. Adv., Available online 23 January 2025, 108522 | https://doi.org/10.1016/j.biotechadv.2025.108522 Cell-free systems: A synthetic biology tool for rapid prototyping in metabolic engineering

Kumyoung Jeung ^a,1, Minsun Kim ^c,1, Eunsoo Jang ^a,1, Yang Jun Shon ^b, Gyoo Yeol Jung ^a,b

^aDivision of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, 77 Cheongam-Ro, Nam-Gu, Pohang, Gyeongbuk 37673, Republic of Korea
^bDepartment of Chemical Engineering, Pohang University of Science and Technology, 77 Cheongam-Ro, Nam-Gu, Pohang, Gyeongbuk 37673, Republic of Korea
^cCenter for Bio-based Chemistry, Korea Research Institute of Chemical Technology (KRICT), 406-30, Jongga-Ro, Jung-Gu, Ulsan 44429, Republic of Korea

¹These authors contributed equally to the work.
Corresponding author : Gyoo Yeol Jung

Abstract

Microbial cell factories provide sustainable alternatives to petroleum-based chemical production using cost-effective substrates. A deep understanding of their metabolism is essential to harness their potential along with continuous efforts to improve productivity and yield. However, the construction and evaluation of numerous genetic variants are time-consuming and labor-intensive. Cell-free systems (CFSs) serve as powerful platforms for rapid prototyping of genetic circuits, metabolic pathways, and enzyme functionality. They offer numerous advantages, including minimizing unwanted metabolic interference, precise control of reaction conditions, reduced labor, and shorter Design-Build-Test-Learn cycles. Additionally, the introduction of in vitro compartmentalization strategies in CFSs enables ultra-high-throughput screening in physically separated spaces, which significantly enhances prototyping efficiency. This review highlights the latest examples of using CFS to overcome prototyping limitations in living cells with a focus on rapid prototyping, particularly regarding gene regulation, enzymes, and multienzymatic reactions in bacteria. Finally, this review evaluates CFSs as a versatile prototyping platform and discusses its future applications, emphasizing its potential for producing high-value chemicals through microbial biosynthesis.

논문정보

형식Research article
게재일2025년 01월 (BRIC 등록일 2025-02-07)
연구진국내 연구진
분야 생명과학 > 생물공학

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