Vibrio　alginolyticus　LHF01　was　engineered　to　efficiently　produce　poly-3-hydroxybutyrate　(PHB)　from　starch　in　this　study.　Firstly,　the　ability　of　Vibrio　alginolyticus　LHF01　to　directly　accumulate　PHB　using　soluble　starch　as　the　carbon　source　was　explored,　and　the　highest　PHB　titer　of　2.06 g/L　was　obtained　in　18 h　shake　flask　cultivation.　Then,　with　the　analysis　of　genomic　information　of　V.　alginolyticus　LHF01,　the　PHB　synthesis　operon　and　amylase　genes　were　identified.　Subsequently,　the　effects　of　overexpressing　PHB　synthesis　operon　and　amylase　on　PHB　production　were　studied.　Especially,　with　the　co-expression　of　PHB　synthesis　operon　and　amylase,　the　starch　consumption　rate　was　improved　and　the　PHB　titer　was　more　than　doubled.　The　addition　of　20 g/L　insoluble　corn　starch　could　be　exhausted　in　6-7 h　cultivation,　and　the　PHB　titer　was　4.32 g/L.　To　the　best　of　our　knowledge,　V.　alginolyticus　was　firstly　engineered　to　produce　PHB　with　the　direct　utilization　of　starch,　and　this　stain　can　be　considered　as　a　novel　host　to　produce　PHB　using　starch　as　the　raw　material.　Copyright　©　2023　Li,　Tian,　Lian,　Fan　and　Li.