Kennedy J F, Barker S A, Pike V W
Biochim Biophys Acta. 1977 Sep 15;484(1):115-26. doi: 10.1016/0005-2744(77)90118-8.
Papain (EC 3.4.22.2) has been coupled to supports of titanium (IV) oxide and cellulose, which are particulate and pre-coated with diazotised 1,3-diaminobenzene, giving water-insoluble and stable derivatives which possess low proteolytic activity but high esterolytic activity. In addition the reversible binding of zinc (II) at the active site of papain has been exploited to inhibit protectively the enzyme during its linkage to the aforementioned supports, thereby yielding water-insoluble derivatives of papain having superior activity upon reactivation with EDTA. Application of the improved procedure of enzyme coupling to macroporous cellulose particles gave a water-insoluble derivative of papain having further enhanced proteolytic activity. Other properties of the water-insoluble derivatives of papain and of similarly prepared water-insoluble conjugates of urease (EC 3.5.1.5) and cholinesterase (EC 3.1.1.8) with cellulose are also reported.
木瓜蛋白酶（EC 3.4.22.2）已与二氧化钛和纤维素载体偶联，这些载体为颗粒状，预先用重氮化的1,3 - 二氨基苯包被，得到水不溶性且稳定的衍生物，其具有低蛋白水解活性但高酯水解活性。此外，木瓜蛋白酶活性位点上锌（II）的可逆结合已被用于在其与上述载体连接过程中对酶进行保护性抑制，从而产生在用乙二胺四乙酸重新激活后具有优异活性的木瓜蛋白酶水不溶性衍生物。将改进的酶偶联程序应用于大孔纤维素颗粒得到了具有进一步增强蛋白水解活性的木瓜蛋白酶水不溶性衍生物。还报道了木瓜蛋白酶水不溶性衍生物以及脲酶（EC 3.5.1.5）和胆碱酯酶（EC 3.1.1.8）与纤维素类似制备的水不溶性缀合物的其他性质。
