Abstract：Type Ⅰ collagen from sheep tendon was prepared by the following sequential steps: degreasing and
decalcification of the starting raw material, removal of impurity proteins, removal of proteoglycans by extraction with
guanidinium hydrochloride, pepsin-catalyzed hydrolysis, salting out with sodium chloride and dialysis. The optimal
conditions for enzymatic hydrolysis were determined by single-factor experiments and orthogonal array design experiment
as hydrolysis at an enzyme concentration of 4% for 60 h in 0.1 mol/L citric acid with a solvent to substrate ratio of 5:1 mL/g. UV
spectral analysis of the resultant extract showed maximum absorption at approximately 220 nm, confirming it to be collagen.
The protein composition of sheep tendon was dominated by type I collagen, as demonstrated by SDS-PAGE analysis. The
extract was further confirmed by infrared spectral analysis to be type Ⅰ collagen with intact structure.
2014, Vol. 28 
