The Stabilizing Mechanism of Immobilized Metagenomic Xylanases on Bio-Based Hydrogels to Improve Utilization Performance: Computational and Functional Perspectives

Bioconjug Chem. 2020 Sep 16;31(9):2158-2171. doi: 10.1021/acs.bioconjchem.0c00361. Epub 2020 Aug 11.

Abstract

While polysaccharide-based superabsorbent hydrogels (SHs) have attracted increasing interest as proficient carriers in the enzyme immobilization, the nature of the favored interactions between the SHs and enzymes is still unclear. Herein, a combined experimental and computational study was employed to investigate the dominant parameters affecting on the stabilization of two metagenomic xylanases on the SHs. The thermostable enzymes (PersiXyn3 and PersiXyn4) with similar domains were screened, cloned, expressed, and purified from cattle rumen metagenome. Then, the enzymes were immobilized on the carboxymethyl cellulose-g-poly(acrylic acid-co-acrylamide) hydrogel which resulted in increasing their activity and stability. The carboxymethyl cellulose (CMC)-based characteristic of the hydrogel provided high numbers of H-bondings/ionic bridges, causing an improvement in the stability, hydrolysis performance, and reusability of the immobilized enzymes. More specifically, enzyme immobilization resulted in ∼40% increase in the content of the reducing sugars released after treatment of paper pulp. After 16 reuse cycles, the immobilized PersiXyn4 displayed 35.9% activity, but the immobilized PersiXyn3 retained just 8.2% of its initial activity. The comparative investigations illustrated that a higher number of positively charged amino acids in the binding site of the enzyme provided stronger electrostatic attractions between it and negative functionalities of the hydrogel. This was suggested as the main reason for the higher affinity of PersiXyn4 toward hydrogel and explained the better hydrolysis performance and reusability of the immobilized PersiXyn4 on the SH. These findings are essential for designing novel innovative SH carriers and the successful engineering of optimal enzyme assemblies through the prediction of the immobilized enzyme's stabilities.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acrylamides / chemistry*
  • Animals
  • Bacteria / chemistry
  • Bacteria / enzymology*
  • Carboxymethylcellulose Sodium / analogs & derivatives*
  • Cattle
  • Endo-1,4-beta Xylanases / chemistry*
  • Enzyme Stability
  • Enzymes, Immobilized / chemistry*
  • Hydrogels / chemistry*
  • Metagenome
  • Models, Molecular

Substances

  • Acrylamides
  • Enzymes, Immobilized
  • Hydrogels
  • poly(acrylamide-co-acrylic acid)
  • Endo-1,4-beta Xylanases
  • Carboxymethylcellulose Sodium