Advances in adaptive laboratory evolution applications for .

Adaptive Laboratory Evolution (ALE), a well-established framework in microbial evolution research, is widely applied in synthetic biology. By simulating natural selection through controlled serial culturing, ALE promotes the accumulation of beneficial mutations, leading to the emergence of specific adaptive phenotypes and bypassing the complexities inherent in rational genetic engineering. With advancements in next-generation sequencing and molecular biology, the integration of high-throughput omics and molecular tools with ALE has significantly enhanced the mapping of genotype-phenotype relationships and the characterization of mutational landscapes. This has propelled progress in microbial evolution, biochemical theory, and interdisciplinary applications. (), a premier chassis in synthetic biology, benefits from its genetic tractability and metabolic flexibility, making it an ideal model for ALE studies. This review examines recent developments in ALE applications for , exploring its methodological principles, experimental design paradigms, notable case studies, and synergies with emerging technologies, providing valuable theoretical insights and practical guidance for related research.