Development of an enzyme-immobilized support using a polyester woven fabric

Abstract

In this study, we aimed to develop an enzyme-immobilized support using polyester woven fabrics and to optimize the development process. We obtained information about the storage stability and reusability of the enzyme and showed the applicability of the polyester woven fabric as an enzyme-immobilized support. In particular, the samples hydrolyzed by hydrogen chloride were treated with N,N'-dicyclohexylcarbodiimide and N-hydroxysuccinimide to activate the surfaces. We evaluated the relative activity of the enzyme immobilization processes, the introduction of spacers, crosslinking and enzyme immobilization and optimized these parameters. The introduction process was controlled to a bovine serum albumin concentration of 1.5% (w/v) and treatment time of 3 h. The crosslinking process was optimized to pH 10.0, a glutaraldehyde concentration of 3% (v/v) and a crosslinking time of 90 min. The immobilization conditions were maintained at pH 8.5, a temperature of 25 C, a time of 45 min and a trypsin concentration of 6% (o.w.f.).Enzyme-immobilized supports have been used in various medical applications, food production processes, ion batteries and filters for waste-water purification. 1,2 Numerous methods for enzyme immobilization on different supports have been described in the literature. Ideal support properties include mechanical stability, hydrophilicity, resistance to microbial attack, porosity and availability at low cost. 2,3 The application of fabrics as immobilization supports could offer several advantages in terms of price and structure compared with other materials such as gels, beads, membranes and films. 2-4 Fabrics are commercially available and relatively cheaper than these or inorganic materials. The fiber-based structures have a high porosity and a large average pore size, so that more enzymes are immobilized, and the immobilized enzymes are protected from the environment. [2][3][4] There are various fabric supports available for enzyme immobilization, which can be classified into natural and synthetic fibers. Natural fibers such as cellulose and chitosan, which have excellent biocompatibility and hydrophilicity, are non-toxic and biodegradable. However, the weak mechanical stability of these natural fibers has greatly limited their application. 3,5 Synthetic fibers such as polylactic acid, polystyrene, polyamide and polyester (polyethylene terephthalate, PET) can be used for enzyme supports due to their good mechanical stability. However, their hydrophobicity is the major disadvantage. It is well known that the hydrophobic interaction between the enzyme and the support is one of the main reasons for protein denaturation. 3,5 One method to address these problems is to introduce spacers between the enzymes and the support. 6 This is a common method to reduce undesirable interactions between the functional groups of support surfaces and the enzyme molecule, and to lower steric hindrance. 7 In particular, the introduction of spacers increases the distance between the enzyme and the hydrophobic surfaces of sy...

A Process for Well-Defined Polymer Synthesis through Textile Dyeing Inspired Catalyst Immobilization

“…Textile dyeing technologies, through which fabrics are imparted with different colors using dyeing chemistry to manufacture the final textile products, provided an inspiring idea for catalyst immobilization. First, fabrics produced from cotton, polyamide, or polyester are promising carriers for catalyst immobilization due to the following advantages: (1) good mechanical properties, excellent durability, high chemical resistance, smooth surfaces, and good processability; , (2) the intrinsic flexibility of fabrics makes them capable of being fitted in any reactor geometry and being separated easily without leaving any residues; (3) pore diffusion, which will influence the reactivity of the catalyst, can also be avoided due to the smooth fabric surface; (4) pendant groups on the surface of the polymeric fabrics, for instance, hydroxyl groups on the surface of cellulosic fabrics, or amino groups on the wool fabrics, are powerful moieties for anchoring catalysts; , (5) manufacturing costs for fabrics are much lower than other solid supports, which is especially true for cotton considered as a nearly inexhaustible natural source for fabric production. , Therefore, a variety of bioactive molecules and organocatalysts have been extensively immobilized on fabrics made of cotton, , wool, polyester, , poly­(ethylene terephthalate), ,, poly­(vinyl alcohol), and poly­(amide 6.6). ,, …”

Protease Immobilization in Solution-Blown Poly(ethylene oxide) Nanofibrous Nonwoven Webs

“…Further experiments would be needed to determine if this hypothesis is valid. When redissolved in buffer, the EFSBN webs showed at least 60% of their initial activity after 300 days, which is higher than the residual activities of immobilized protease in various polymeric supports reported in the literature as only 33–57% after a much shorter storage period of 30 days at 4 °C. ,, This result revealed that immobilized protease in EFSBN could be conveniently stored at ambient temperature or in refrigerators. The produced EFSBN are a unique storage technique in a quick-dissolving solid form rather than typical liquid enzymes or lyophilized forms.…”

“…When redissolved in buffer, the EFSBN webs showed at least 60% of their initial activity after 300 days, which is higher than the residual activities of immobilized protease in various polymeric supports reported in the literature as only 33−57% after a much shorter storage period of 30 days at 4 °C. 17,65,66 This result revealed that immobilized protease in EFSBN could be conveniently stored at ambient temperature or in refrigerators.…”

Immobilization and stabilization of alcohol dehydrogenase on polyvinyl alcohol fibre

“…Several studies have reported the effect of spacer length and type of spacer between enzyme and carrier and its effect on the activity of the immobilized enzyme [ 38 , 46 , 47 ]. Higher activity is observed when a spacer is incorporated between the carrier and enzyme to reduce steric hindrance, promote conformational mobility, and reduce adverse surface interaction [ 18 , 19 , 48 ]. A spacer was introduced to the PVA-Cl discs by reaction with diamine in ethanol and resultant amine was reacted with glutaraldehyde [ 49 , 50 ].…”