<p>Antibiotic resistance has become a public safety issue of the twenty-first century, posing a growing threat and drawing increased attention. Compared to traditional antibiotics, antimicrobial peptides (AMPs), as naturally produced small peptides, can target multiple pathways within pathogens and render them less prone to developing resistance. This makes them promising alternatives to antibiotics. However, traditional chemical synthesis methods face challenges, such as high costs, low yields, and poor stability, limiting the large-scale industrial production of AMPs. Despite extensive research to improve AMP production efficiency, issues such as low yields and complex extraction processes continue to pose significant barriers to commercial application. Therefore, there is an urgent need for new biosynthesis strategies and optimization methods to enhance AMP production efficiency and quality. This review summarizes the sources, classification, mechanisms of action and recent advances in the microbial synthesis of AMPs. It also explores innovative production methods, including recombinant microbial expression systems, fusion tags, codon optimization, tandem multimer expression, and hybrid peptide expression. Furthermore, we review the applications of gene editing technologies and artificial intelligence in AMP production, providing new perspectives and strategies for efficient, large-scale AMP production.</p> Graphical Abstract <p></p>