羊软骨中Ⅱ型胶原蛋白肽的制备和抑菌活性

Preparation and antioxidant antivity of type Ⅱ collegen peptides from sheep cartilage

以羊软骨胶原蛋白为原料,利用中性蛋白酶、碱性蛋白酶、胃蛋白酶、胰蛋白酶和木瓜蛋白酶对其进行水解,确定最合适的酶解时间和酶用量以水解液的抑菌率和水解度为指标,比较酶解效果;以大肠杆菌为指示菌,通过双层琼脂平板扩散法检测其抑菌活性。试验表明:碱性蛋白酶酶解羊软骨胶原蛋白得到的水解液的水解度最高,最合适的酶解时间是2h,酶用量(占胶原蛋白的百分比)4%;碱性蛋白酶酶解羊软骨胶原蛋白得到的水解液的抑菌率最大,最合适的酶解时间是2.5 h,酶用量(占胶原蛋白的百分比)4%,水解液对大肠杆菌有抑菌活性,其抑菌圈直径为(26.79±0.16)mm,最后得到的胶原蛋白肽对大肠杆菌、金黄色葡糖球菌、枯草芽孢杆菌、荧光假单胞菌和沙门氏菌都有抑菌作用。

The sheep cartilages collagen were collected as raw materials,five different proteases,neutral protease,alkaline protease,pepsin,trypsin and papain,were compared for their effectiveness in hydrolyzing sheep cartilages collagen.Antibacterial activityand degree of hydrolysis as an index hydrolyzate.E.coli as indicator bacteria,antibacterial activity was detected by the double agar diffusion method.The result showed that alkaline protease was found to provide maximum degree of hydrolysis among the proteases and the optimal conditions for hydrolyzing sheep cartilages collagen with it were determined as 4% of enzyme dosage（relative to the weight of collagen） and 2 h of hydrolysis time.The hydrolysates obtained using alkaline protease had an inhibitory effect on Escherichia coli with an inhibition zone diameter of 27.69 mm and the optimal conditions for hydrolyzing sheep cartilages collagenwith it were determined as 4% of enzyme dosage（relative to the weight ofcollagen） and 2 h of hydrolysis time.The resulting collagen peptides on Escherichia coli,Staphylococcus aureus,Bacillus subtilis,Pseudomonas fluorescens,Salmonella enterica subsp.enterica have antimicrobial effect.