Enzyme and Microbial Biochemistry Laboratory, Department of Chemistry, Indian Institute of Technology Delhi, New Delhi 110016, India.
Enzyme Technology Laboratory, Department of Biosciences, Jamia Millia Islamia, New Delhi 110025, India.
Int J Biol Macromol. 2020 Nov 15;163:1747-1758. doi: 10.1016/j.ijbiomac.2020.09.091. Epub 2020 Sep 19.
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.
化学交联剂难以从支架材料中去除，这限制了它们在组织工程中的应用；设计高效的生物相容性水凝胶虽然具有挑战性，但却是理想的选择。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（MWCNTs）上，以将其应用于水凝胶支架设计中。MTGase 来自无毒的生物交联剂摩拉氏链霉菌，采用更环保的方法，具有增强的生化和结构特性。当 MTGase 通过共价键偶联到 MWCNTs 上时，实现了 58%的最大固定化效率。动力学研究表明，催化效率提高了 4.76 倍，可重复使用 7 次以上。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表现出良好的交联效率，导致水凝胶的溶胀比降低。我们的研究结果首次报道了将固定化 MTGase 到 MWCNTs 上的新型生物相容性水凝胶支架的开发。水凝胶在应用过程中不可避免地会受到损伤。为了弥补水凝胶的损伤，创建仿生水凝胶以模拟天然组织微环境具有重要意义。微生物 TGase 具有广阔的应用前景，可作为组织工程中水凝胶支架的交联剂。
