以前期成功构建出降解胶原蛋白的工程菌pET30a-ColM13/BL21为出发菌，对工程菌表达纯化出的重组胶原酶(ColM13)的酶学性质进行分析。通过研究温度、pH、抑制剂对其的影响，得出ColM13的最适反应温度为50 ℃，最适反应pH为8.0；在20～55 ℃和pH6.0～9.0范围内有良好的稳定性；金属蛋白酶抑制剂乙二胺四乙酸(ethylenediaminetetraacetic acid,EDTA)、乙二醇双(2-氨基乙基醚)四乙酸(ethylenebis(oxyethylenenitrilo tetraacetic acid, EGTA)对ColM13有显著的抑制作用。采用紫外光谱(ultraviolet and visible spectrum, UV)、差示量热扫描(differential scanning calorimeter, DSC)、荧光光谱(FS)、傅里叶红外光谱(FT-IR)对骨胶原蛋白及其酶解产物进行结构特性分析，分析可知酶解作用使骨胶原蛋白的三股螺旋结构遭到破坏，释放出大量的游离氨基酸。酶解后胶原蛋白的肽链上相互排斥减弱，疏水氨基酸残基作用增强，具有更高的热稳定性。与B.cereus MBL13-U分离纯化出的胶原酶(BSC)相比，ColM13处理的胶原蛋白的降解效果更明显。

In this study, the enzymatic properties of the recombinant collagenase (ColM13) from engineering bacteria pET30a-ColM13/BL21 were analyzed after expression and purification.By studying the effects of temperature, pH and inhibitor, the optimal reaction temperature and pH for the ColM13 enzyme activity were determined as 50 ℃ and 8.0 respectively.The ColM13 enzyme was stable when the temperature was 20-55 ℃.The ColM13 enzyme activity was stable between pH 6.0-9.0.Metalloproteinase inhibitors EDTA and EGTA had a significant inhibitory effect on ColM13.The structural properties of bone collagen and its hydrolysate were characterized by ultraviolet (UV), differential scanning calorimetry (DSC), fluorescence and fourier transform infrared spectroscopy (FT-IR).The results showed that the enzymatic hydrolysis damaged the triple helical structure of collagen protein and released a large amount of free amino acids.The enzymolysis reduced the -NH2+- mutual exclusion on the collagen peptide chain and enhanced the hydrophobic amino acid residues.The collagen hydrolysate had higher thermal stability.Compared with the collagenase (BSC) isolated from B.cereus MBL13-U, the degradation of collagen treated by ColM13 was more significant.