Enhancing the Thermo-Stability and Anti-Biofilm Activity of Alginate Lyase by Immobilization on Low Molecular Weight Chitosan Nanoparticles

Abstract

Bacterial biofilm causes severe antibiotic resistance. An extracellular polymeric substance (EPS) is the main component in the bacterial biofilm. Alginate is a key EPS component in the biofilm of Pseudomonas aeruginosa and responsible for surface adhesion and stabilization of biofilm. Alginate lyase has emerged as an efficient therapeutic strategy targeting to degrade the alginate in the biofilm of P. aeruginosa. However, the application of this enzyme is limited by its poor stability. In this study, chitosan nanoparticles (CS-NPs) were synthesized using low molecular weight chitosan and alginate lyase Aly08 was immobilized on low molecular weight chitosan nanoparticles (AL-LMW-CS-NPs). As a result, the immobilization significantly enhanced the thermal stability and reusability of Aly08. In addition, compared with free Aly08, the immobilized AL-LMW-CS-NPs exhibited higher efficiency in inhibiting biofilm formation and interrupting the established mature biofilm of P. aeruginosa, which could reduce its biomass and thickness confirmed by confocal microscopy. Moreover, the biofilm disruption greatly increased the antibiotic sensitivity of P. aeruginosa. This research will contribute to the further development of alginate lyase as an anti-biofilm agent.

Eradication of Acinetobacter baumannii Planktonic and Biofilm Cells Through Erythrosine-Mediated Photodynamic Inactivation Augmented by Acetic Acid and Chitosan

“…In this study, chitosan enhanced the anti-biofilm efficacy of erythrosine B-mediated PDI against A. baumannii . Enhancement of the efficacy of EB-mediated PDI using chitosan might be attributed to the following three reasons: (1) the ability of chitosan to disrupt the biofilm structure and thus helping the PDI to act on the cells that are released from the biofilm, (2) the ability of chitosan to permeabilize the outer membrane of gram-negative bacteria such as A. baumannii , and (3) the ability of cationic chitosan to act as a drug carrier for delivery of erythrosine to biofilm [ 12 , 31 , 32 ].…”

“…Acetic acid, an organic chemical compound, can modify the molecular charge of some photosensitizers under acidic condition that subsequently affect their activity during PDI [ 10 ]. Chitosan is a natural polycationic antimicrobial biopolymer known to be able to destabilize the biofilm architecture [ 11 , 12 ].…”

Alpha‐lipoic acid improves the reproduction performance of breeder hens during the late egg‐laying period

“…In this study, 600 mg/kg ALA-treated group decreased the level of MDA and improved the activity of TAOC and SOD in ovary. Previous studies have noted that ALA can enhance the antioxidants enzymes like superoxide dismutase, by reducing the substrate and free radicals to promote the lipid stability in birds (Kofuji, Nakamura, Nakamura, Isobe, Murata, & Kawashima, 2008;Li et al, 2019 (Liu, Lin, Mi, Li, et al, 2018;Liu, Lin, Mi, Zeng, et al, 2018). ALA has been reported to increase the reproductive hormones in ovarian oxidative damage rats (Nair et al, 2020).…”

“…In this study, 600 mg/kg ALA‐treated group decreased the level of MDA and improved the activity of TAOC and SOD in ovary. Previous studies have noted that ALA can enhance the antioxidants enzymes like superoxide dismutase, by reducing the substrate and free radicals to promote the lipid stability in birds (Kofuji, Nakamura, Nakamura, Isobe, Murata, & Kawashima, 2008; Li et al, 2019). Furthermore, ALA, as a coenzyme of mitochondria, can alleviate the loss of mitochondrial function due to ageing and reduce oxidative phosphorylation of the ovary (Di Tucci et al, 2018), thus improving the reproductive pathway.…”

Enhancing the Thermo-Stability and Anti-Bacterium Activity of Lysozyme by Immobilization on Chitosan Nanoparticles

“…Tarhan et al prepared maltose-functionalized magnetic core-shell nanoparticles (Fe3O4@Au NPs) to effectively immobilized L-asparaginase (L-ASNase) and characterized their properties, being significantly improved acid-base tolerance and thermal stability and retention of 50% relative activity after reusing 13 cycles [27]. In addition, we also successfully immobilized a polysaccharide degrade enzyme, alginate lyase Aly08, into low molecule weight chitosan nanoparticles, which shows a great thermal stability and reusability compared to free Aly08 [28]. In this study, enzyme immobilization stabilized the enzyme conformation and helped to resist thermal denaturation and leaching, while the immobilized lysozyme showed better thermal stability and superb reusability.…”