Multienzyme Cellulose Films as Sustainable and Self-Degradable Hydrogen Peroxide-Producing Material

Abstract

The use of hydrogen peroxide-releasing enzymes as a component to produce alternative and sustainable antimicrobial materials has aroused interest in the scientific community. However, the preparation of such materials requires an effective enzyme binding method that often involves the use of expensive and toxic chemicals. Here, we describe the development of an enzyme-based hydrogen peroxide-producing regenerated cellulose film (RCF) in which a cellobiohydrolase (TrCBHI) and a cellobiose dehydrogenase (MtCDHA) were efficiently adsorbed, 90.38 ± 2.2 and 82.40 ± 5.7%, respectively, without making use of cross-linkers. The enzyme adsorption kinetics and binding isotherm experiments showed high affinity of the proteins possessing cellulose-binding modules for RCF, suggesting that binding on regenerated cellulose via specific interactions can be an alternative method for enzyme immobilization. Resistance to compression and porosity at a micrometer scale were found to be tunable by changing cellulose concentration prior to film regeneration. The self-degradation process, triggered by stacking TrCBHI and MtCDHA (previously immobilized onto separate RCF), produced 0.15 nmol/min·cm2 of H2O2. Moreover, the production of H2O2 was sustained for at least 24 h reaching a concentration of ∼2 mM. The activity of MtCDHA immobilized on RCF was not affected by reuse for at least 3 days (1 cycle/day), suggesting that no significant enzyme leakage occurred in that timeframe. In the material herein designed, cellulose (regenerated from a 1-ethyl-3-methylimidazolium acetate/dimethyl sulfoxide (DMSO) solution) serves both as support and substrate for the immobilized enzymes. The sequential reaction led to the production of H2O2 at a micromolar–millimolar level revealing the potential use of the material as a self-degradable antimicrobial agent.

Degradation of Cellulose Derivatives in Laboratory, Man-Made, and Natural Environments

“…However, as other studies have demonstrated that UV irradiation in combination with H 2 O 2 can trigger rapid self-destruction of cellulose materials, this approach could possibly also be utilized for the preparation of self-degradable cellulose films. 55 …”

“…H 2 O 2 producing enzymes, cellobiohydrolase and cellobiosedehydrogenase, have also been adsorbed on cellulose films aimed at antibacterial surfaces. However, as other studies have demonstrated that UV irradiation in combination with H 2 O 2 can trigger rapid self-destruction of cellulose materials, this approach could possibly also be utilized for the preparation of self-degradable cellulose films …”

Alignment–Rheology Relationship of Biosourced Rod-Like Colloids and Polymers under Flow

“…Biosourced rod-like colloids (RC) and rod-like polymers (RP) have excellent mechanical and optical properties and are often regarded as sustainable and biocompatible building blocks. − These features have been exploited to form novel materials − with applications in biomedical − and food engineering. − Classical examples of biosourced RC that can be produced in industrially relevant quantities comprise cellulose nanofibrils, cellulose nanocrystals, and protein nanofibrils. Nonetheless, these RC are characterized by a polydisperse contour length which complicates the prediction of their flow behavior.…”

Study of CAZymes from >i<Thermothelomyces thermophilus>/i< M77 as antibiofilm agents: an oxidative and hydrolytic approach

“…This approach enables us to continuously produce H2O2 without the need for external addition of cellobiose or glucose, achieving up to 170 μM which is higher that our previously reported quantity of 90 μM, using TthCDHIIa with cellobiose (Figure 25c). Similar approaches using celluloses disc chemically synthesized have been reported 39 .…”

“…In the last two decades a growth in infections caused by drug-resistant and biofilmforming bacteria has been consistently reported 37,38 . CDH might play a role as an attractive antimicrobial and antibiofilm agent, due to its ability to continuously produce hydrogen peroxide from oligosaccharides like cellobiose, 39 which are not metabolizable by pathogens 40 . This non-canonical use of CDH and other enzymes in biomedical applications has been growing, as documented by several recent studies [41][42][43][44][45] .…”