Collagenase Ⅱ is a matrix metalloproteinase (MMPs) used for tissue dissociation and the preparation of liver, bone, thyroid, heart, and salivary gland cells[1,2]. Collagenase can cleave the bond between neutral amino acids and glycine in the Pro-X-Glyc-Pro sequence, which is common in collagen [3]. The enzyme activity of collagenase requires the presence of divalent cations such as Ca2+ and Zn2+ for activation[4]. Collagenase activity is inhibited by agents such as ethylenediaminetetraacetic acid (EDTA), dithiothreitol (DTT), detergents, hexachlorocyclohexane, and heavy metal ions (Hg2+, Pb2+, Cd2+) [5]. Collagenase is relatively mild and exhibits good dissociation ability at physiological temperature and pH without the need for mechanical stirring or special equipment. The optimal pH for Collagenase II is 6.3-8.5, and its enzymatic activity is ≥125 CDU/mg solid (CDU = Collagen Digestion Units).
There are approximately five common types of collagenase [6]:
(1) Collagenase Type I (GC19589): Contains relatively uniform multiple enzymatic activities (including collagenase, caseinase, clostridiopeptidase, and trypsin activities), and is commonly used for the dissociation of epithelial, lung, adipose, and adrenal tissue cells.
(2) Collagenase Type II (GC19588): Contains higher clostridiopeptidase activity and is typically used for the dissociation of liver, bone, thyroid, heart, and salivary gland tissue cells.
(3) Collagenase Type III (GC19590): Contains lower protease activity and is commonly used for the dissociation of mammary gland cells.
(4) Collagenase Type IV (GC19591): Contains lower trypsin activity and is typically used for the preparation of islet cells or experiments that require the preservation of cell surface receptors.
(5) Collagenase Type V (GC19592): Partially purified, containing higher collagenase and caseinase activities, and is typically used for the dissociation of pancreatic islet tissue and connective tissue dissociation.
References:
[1] Sekhon B S. Matrix metalloproteinases–an overview[J]. Research and Reports in Biology, 2010: 1-20.
[2] Alipour H, Raz A, Zakeri S, et al. Therapeutic applications of collagenase (metalloproteases): A review[J]. Asian Pacific Journal of Tropical Biomedicine, 2016, 6(11): 975-981.
[3] Trabelsi O, Dumas V, Breysse E, et al. In vitro histomechanical effects of enzymatic degradation in carotid arteries during inflation tests with pulsatile loading[J]. Journal of the mechanical behavior of biomedical materials, 2020, 103: 103550.
[4] Eming S, Smola H, Hartmann B, et al. The inhibition of matrix metalloproteinase activity in chronic wounds by a polyacrylate superabsorber[J]. Biomaterials, 2008, 29(19): 2932-2940.
[5] Daboor S M, Budge S M, Ghaly A E, et al. Extraction and purification of collagenase enzymes: a critical review[J]. Am. J. Biochem. Biotechnol, 2010, 6(4): 239-263.
[6] Alipour H, Raz A, Zakeri S, et al. Therapeutic applications of collagenase (metalloproteases): A review[J]. Asian Pacific Journal of Tropical Biomedicine, 2016, 6(11): 975-981.
Collagenase Ⅱ是一种基质金属蛋白酶（MMPs），用于组织分离，制备肝、骨、甲状腺、心脏和唾液腺细胞[1,2]。胶原酶可以切割 Pro-X-Glyc-Pro 序列中的中性氨基酸与甘氨酸之间的键，这些序列在胶原蛋白中很常见[3]。胶原酶需要借助二价阳离子如Ca2+、 Zn2+等活化酶的活性[4]。胶原酶的活性会受到乙二胺四乙酸（EDTA）、二硫苏糖醇（DTT）、洗涤剂、六氯环己烷和重金属离子（Hg2+、Pb2+、Cd2+）等的抑制作用[5]。胶原酶相对温和，在生理温度和生理pH值条件下具有良好的解离能力，无需机械搅拌或特殊设备。II型胶原酶的最适pH为6.3-8.5，酶活力≥125 CDU/mg固体（CDU =胶原蛋白消化单位）。
常用胶原酶大约有以下5种类型[6]：
（1）I型胶原酶（GC19589）含有相对均匀的多种酶活性（包括胶原酶、酪蛋白酶、梭菌蛋白酶、和胰蛋白酶活性），通常用于上皮、肺，脂肪和肾上腺组织细胞的分离。
（2）Ⅱ型胶原酶（GC19588）含有更高的梭菌蛋白酶活性，通常用于肝脏，骨，甲状腺，心脏和唾液腺组织细胞的分离。
（3）Ⅲ型胶原酶（GC19590）含有较低的蛋白酶活性，通常用于乳腺细胞的分离。
（4）Ⅳ型胶原酶（GC19591）含有较低的胰蛋白酶活性，通常用于胰岛细胞的制备，或需要保持受体完整性的细胞制备实验。
（5）Ⅴ型胶原酶（GC19592）部分纯化，含有较高的胶原酶和酪蛋白酶活性。通常用于胰腺小岛组织的分离，结缔组织分离。