Collagenase V

Collagenase V is a proteolytic enzyme that is commonly used to digest extracellular matrix proteins and is suitable for the separation of pancreatic islet cells and connective tissue cells^[1]. Collagenase V has higher collagenase and casein activities and lower pancreatic enzyme levels to reduce damage to cell membrane proteins^[1]. Collagenase can cleave the bond between neutral amino acids and glycine in the Pro-X-Glyc-Pro sequence, which is common in collagen^[2]. Collagenase requires the activation of divalent cations such as Ca²⁺ and Zn^{2+ [3]}. Collagenase activity is inhibited by ethylenediaminetetraacetic acid (EDTA), dithiothreitol (DTT), detergents, hexachlorocyclohexane, and heavy metal ions (Hg²⁺, Pb²⁺, Cd²⁺)^[4]. Collagenase is relatively mild and has good dissociation ability under physiological temperature and physiological pH conditions, without the need for mechanical stirring or special equipment. This product is derived from Bacillus histolyticus, with an optimum pH of 7-8 and an enzyme activity ≥125 CDU/mg solid (CDU = collagen digestion unit).

Activity definition: At pH 7.5, 25°C, in the presence of calcium ions, one unit of di-GPA hydrolysis will hydrolyze 1.0μM furanyl acryloyl-Leu-Gly-Pro-Ala per minute. One unit of neutral protease will hydrolyze casein to produce a color equivalent to 1.0μM tyrosine per 5 hours at pH 7.5 and 37°C. At pH 7.6, 25°C, in the presence of DTT, each unit of clostripain will hydrolyze 1.0μM BAEE per minute.

References:
[1] Miyazaki Y, Murayama K, Fathi I, et al. Strategy towards tailored donor tissue-specific pancreatic islet isolation[J]. PLoS One, 2019, 14(5): e0216136.
[2] 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.
[3] 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.
[4] 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.

Collagenase V是一种蛋白水解酶，常用于消化细胞外基质蛋白，适用于胰腺小岛细胞和结缔组织细胞的分离^[1]。Collagenase V具有更高的胶原酶活性和酪蛋白酶活性，更低的胰酶水平以降低对细胞膜蛋白的损伤^[1]。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^[2]。胶原酶需要借助二价阳离子如Ca²⁺、 Zn²⁺等活化酶的活性^[3]。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^[4]。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。本产品来源于溶组织棱状芽胞杆菌，最适pH为7-8，酶活力≥125 CDU/mg固体（CDU =胶原蛋白消化单位）。

活力定义：在pH 7.5、25 °C和钙离子存在下，一个双GPA水解单元每分钟水解1.0μM呋喃基丙烯酰基-Leu-Gly-Pro-Ala。一个中性蛋白酶单位水解酪蛋白，在pH 7.5和37℃下每5小时产生相当于1.0μM酪氨酸的颜色。在pH 7.6、25℃、DTT存在下，每单位梭菌蛋白酶每分钟水解1.0μM BAEE。