The high porosity, interconnected pore structure, and
high surface
area-to-volume ratio make the hydrophilic nonwoven nanofiber membranes
(NV-NF-Ms) promising nanostructured supports for enzyme immobilization
in different biotechnological applications. In this work, NV-NF-Ms
with excellent mechanical and chemical properties were designed and
fabricated by electrospinning in one step without using additives
or complicated crosslinking processes after electrospinning. To do
so, two types of ultrahigh-molecular-weight linear copolymers with
very different mechanical properties were used. Methyl methacrylate-
co
-hydroxyethyl methacrylate (p(MMA)-
co
-p(HEMA)) and methyl acrylate-
co
-hydroxyethyl acrylate
(p(MA)-
co
-p(HEA)) were designed and synthesized by
reverse atom transfer radical polymerization (reverse-ATRP) and copper-mediated
living radical polymerization (Cu
0
-MC-LRP), respectively.
The copolymers were characterized by nuclear magnetic resonance (
1
H-NMR) spectroscopy and by triple detection gel permeation
chromatography (GPC). The polarity, topology, and molecular weight
of the copolymers were perfectly adjusted. The polymeric blend formed
by (MMA)
1002
-
co
-(HEMA)
1002
(
M
w
= 230,855 ± 7418 Da;
M
n
= 115,748 ± 35,567 Da; PDI = 2.00) and (MA)
11709
-
co
-(HEA)
7806
(
M
w
= 1.972 × 10
6
± 33,729 Da;
M
n
= 1.395 × 10
6
± 35,019
Da; PDI = 1.41) was used to manufacture (without additives or chemical
crosslinking processes) hydroxylated nonwoven nanofiber membranes
(NV-NF-Ms-OH; 300 nm in fiber diameter) with excellent mechanical
and chemical properties. The morphology of NV-NF-Ms-OH was studied
by scanning electron microscopy (SEM). The suitability for enzyme
binding was proven by designing a palette of different surface functionalization
to enable both reversible and irreversible enzyme immobilization.
NV-NF-Ms-OH were successfully functionalized with vinyl sulfone (281
± 20 μmol/g), carboxyl (560 ± 50 μmol/g), and
amine groups (281 ± 20 μmol/g) and applied for the immobilization
of two enzymes of biotechnological interest. Galactose oxidase was
immobilized on vinyl sulfone-activated materials and carboxyl-activated
materials, while laccase was immobilized onto amine-activated materials.
These preliminary results are a promising basis for the application
of nonwoven membranes in enzyme technology.