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,　safe,　high　activity　antifreeze　peptides　using　various　separating　means,　and　to　detect　their　hypothermia　protect　activity　on　Lactobacillus　bulgaricus.　Collagen　derived　from　shark　skin　was　hydrolyzed　with　Acid　Protease.　Then　the　hydrolysate　was　subsequently　added　on　to　SP-Sephadex　C-25　column,　Sephadex　G-50　gel　filtration　column　and　C18　Reversed-phase　high　performance　liquid　chromatography　to　acquire　a　pure　component.　A　purified　hydrophilic　peptide,　named　SsC-AFP,　with　the　characteristic　of　pI＜5.0　and　molecular　weight　of　906Da,　was　obtained.　The　amino　acid　sequence　of　SsC-AFP　was　GAIGPAGPLGP,　which　consisted　of　Gly-Pro-x　tripeptide　repeat　sequence.　When　the　concentration　of　SsC-AFP　was　250μg/mL,　the　survival　rate　of　L.　bulgaricus　was　up　to　90.28%.　©　2013　Elsevier　Ltd.