Abstract:　Antifreeze　peptides　can　protect　living　organisms　from　low　temperatures　by　preventing　damage　or　killing　due　to　ice　crystal　formation　between　cells.　Therefore,　antifreeze　peptides　can　be　used　as　a　low　temperature　protectant　for　cryopreservation　of　cells　and　tissues,　and　also　in　food　production.　In　this　study,　a　recombinant　SF-P　gene　was　constructed　and　inserted　into　pNZ8149　to　construct　a　food　grade　expression　vector,　which　was　then　electroporated　into　Lactococcus　lactis　NZ3900.　The　expression　of　the　target　protein　was　induced　using　Nisin,　and　the　optimal　expression　condition　was　determined　to　be　a　pH　of　6.0,　Nisin　concentration　of　25　ng/mL,　temperature　of　37　°C,　and　incubation　time　of　6　hr.　Compared　to　the　strain　NZ3900　and　the　recombinant　strain　SF-P1　without　addition　of　Nisin,　the　recombinant　strain　SF-P2　showed　the　highest　cell　survival　and　thermal　hysteresis　activity,　and　had　a　reduction　in　the　changes　of　activities of　extracellular　and　intracellular　lactate　dehydrogenase　and　β-galactosidase　after　freezing.　Moreover,　analysis　by　SEM　showed　that　SF-P2　cells　were　more　completely　and　regularly　shaped　than　other　strains,　displayed　no　obvious　leakage　of　cell　contents,　and　had　an　intact　boundary　between　cells　after　freezing.　These　results　indicate　that　the　recombinant　strain　SF-P2　has　a　protective　effect　against　freezing.　This　paper　presents　a　food　grade　expression　system　for　an　antifreeze　peptide　SF-P　using　L.　lactis　as　a　host,　and　shows　that　the　intracellular　expression　of　antifreeze　peptide　could　protect　the　cellular　integrity　and　physiological　functions　of　L.　lactis.　Practical　Application:　The　recombinant　Lactococcus　lactis　with　intracellular　expression　of　antifreeze　peptides　SF-P　could　reduce　the　damage　of　bacteria　cells　induced　by　freezing　or　freeze　drying,　so,　it　could　be　applied　in　the　process　of　freezing　food　without　separation,　such　as　the　manufacture　of　yoghurt　ice　cream,　frozen　dough,　and　so　on.　©　2018　Institute　of　Food　Technologists®