Antifreeze　proteins　have　characteristics　of　inhibiting　the　growth　of　crystals,　decreasing　the　injury　of　cells　and　can　retain　the　structure,　texture　and　quality　of　productions.　The　purpose　of　this　study　is　to　obtain　natural　antifreeze　peptides,　and　to　investigate　the　hypothermia　protection　activity　on　bacteria.　Gelatin　derived　from　shark　skin　was　hydrolysed　to　obtain　antifreeze　peptides.　The　most　appropriate　protease　and　hydrolysis　time　was　selected　with　the　index　of　the　hypothermia　protection　activity　on　bacteria.　The　hydrolysate　was　subsequently　added　on　to　Sephadex　G-50　gel　filtration　column　and　SP-Sephadex　C-25　column　to　acquire　high　activity　fractions.　The　fraction　of　cationic　peptides　termed　P2　shows　higher　antifreeze　activity.　The　hypothermia　protection　assay　shows　that　the　survival　rate　of　E.COLI　was　80.8　%　when　the　concentration　of　peptides　complexes　was　up　to　500　μg/mL.