高效合成5-HV工程菌的构建与选育OA

5-Hydroxyvaleric acid is a bio-based platform compound for the synthesis of δ-valerolactone and high-performance biode-gradable polymers,and is of significant importance in the fields of green chemistry and novel materials.Constructing a genetically stable and high-yield strain is essential for the efficient biosynthesis of 5-hydroxyvalerate.In this study,a recombinant strain,DX-JN,was con-structed using high-throughput screening markers,and the relationship between 5-hydroxyvaleric acid production and fluorescence intensity was established.Engineered strains producing 5-hydroxyvaleric acid were screened based on fluorescence intensity through a combination of ultraviolet(UV)mutagenesis and flow cytometry.Using this method,a high-yield 5-hydroxyvaleric acid-producing Escherichia coli strain,DXMCJN6-1,was obtained.A final yield of 15.99 g/L of 5-hydroxyvaleric acid was achieved when 30 g/L of glucose was consumed,cor-responding to a glucose conversion rate of 53.3％.In contrast,the control strain DX-JN synthesized 8.71 g/L 5-hydroxyvaleric acid with a glucose conversion rate of 29％.Notably,the 5-hydroxyvaleric acid yield was significantly increased following high-throughput screening.Overall,both acid production and fermentation performance of the strain were enhanced through high-throughput strain breeding.