The chemical cross-linkers are difficult to be removed from the scaffold materials, which limit their application in tissue engineering; designing an efficient biocompatible hydrogel is though challenging is desirable. The aim of the present study was to immobilize microbial Transglutaminase (MTGase) enzyme on multi-walled carbon nanotubes (MWCNTs) for its application in hydrogel scaffolds designing. MTGase from Streptomyces mobaraensis, a non-toxic biological cross-linker, was employed for a greener approach with enhanced biochemical and structural properties. The maximum immobilization efficiency of 58% was achieved when MTGase was covalently coupled on MWCNTs. The kinetic studies showed 4.76-fold increase in catalytic efficiency and good reusability upto seven cycles. Attachment of enzyme on MWCNTs surface was studied through SEM and FTIR. The immobilized enzyme showed good cross-linking efficiency in gelatin hydrogel scaffold resulting decrease in swelling ratio of hydrogel. Our findings report for the first time the development of novel biocompatible hydrogel scaffolds with immobilized MTGase onto MWCNTS. Inevitable damage of hydrogels are incurred during their applications. To offset the damage of hydrogels, the creation of bioinspired hydrogels emulating native tissue microenvironment is highly significant. Microbial TGase holds promising future with its applicability as a cross-linker of hydrogel scaffolds in the area of tissue engineering.

化学交联剂很难从支架材料中去除，这限制了它们在组织工程中的应用。设计有效的生物相容性水凝胶虽然具有挑战性，但仍是需要的。本研究的目的是将微生物转谷氨酰胺酶（MTGase）固定在多壁碳纳米管（MWCNT）上，以用于水凝胶支架设计。来自茂原链霉菌的MTGase，一种无毒的生物交联剂，被用于绿色方法，具有增强的生化和结构特性。当MTGase共价偶联在MWCNT上时，最大固定效率达到58％。动力学研究表明，在七个周期内，催化效率提高了4.76倍，并且具有良好的可重复使用性。通过SEM和FTIR研究了酶在MWCNTs表面的附着。固定化酶在明胶水凝胶支架中显示出良好的交联效率，导致水凝胶的溶胀率降低。我们的研究结果首次报道了将MTGase固定在MWCNTS上的新型生物相容性水凝胶支架的开发。水凝胶在使用过程中不可避免地会受到损害。为了抵消水凝胶的损害，模拟天然组织微环境的生物启发水凝胶的创建具有重大意义。微生物TGase作为水凝胶支架的交联剂在组织工程领域的应用前景广阔。