Enhancing the stability and antibiofilm activity of DspB by immobilization on carboxymethyl chitosan nanoparticles

• Published in: Microbiological research Vol. 178; pp. 35 - 41
• Main Authors: Tan, Yulong, Ma, Su, Liu, Chenguang, Yu, Wengong, Han, Feng
• Format: Journal Article
• Language: English
• Published: Germany Elsevier GmbH 01.09.2015
• Subjects: Acetylglucosaminidase - chemistry; Acetylglucosaminidase - metabolism; Aggregatibacter actinomycetemcomitans - enzymology; Anti-Infective Agents - chemistry; Anti-Infective Agents - metabolism; Biofilm; Biofilms - drug effects; Carboxymethyl chitosan; Chitosan - analogs & derivatives; Chitosan - metabolism; Cloning, Molecular; Drug Storage; Enzyme Stability; Enzymes, Immobilized - chemistry; Enzymes, Immobilized - metabolism; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Gene Expression; Immobilization; Nanoparticles; Nanoparticles - metabolism; Stability; Staphylococcus aureus; Staphylococcus aureus - drug effects; Staphylococcus aureus - physiology; Staphylococcus epidermidis; Staphylococcus epidermidis - drug effects; Staphylococcus epidermidis - physiology; Temperature; Nanoparticles; Immobilization; Biofilm; Stability; Carboxymethyl chitosan
• ISSN: 0944-5013; 1618-0623
• DOI: 10.1016/j.micres.2015.06.001

A β-N-acetyl-glucosaminidase (DspB) from Aggregatibacter actinomycetemcomitans CU1000 has been proved to inhibit and detach the biofilms formed by Staphylococcus epidermidis, Staphylococcus aureus and A. actinomycetemcomitans. However, the application of this enzyme is limited by its poor stability. In the present study, a β-N-acetyl-glucosaminidase encoding gene, dspB, was cloned from A. actinomycetemcomitans HK1651 and expressed in Escherichia coli. The recombinant DspB was loaded on hydrogel nanoparticles, which was prepared by using linoleic acid (LA) modified carboxymethyl chitosan (CMCS) after sonication. The nanoparticles were almost saturated by DspB at 0.3mg/ml, which gave a loading capacity of 76.7%. The immobilization enhanced thermal stability, storage stability and reusability of DspB significantly. Moreover, it also increased antibiofilm activity due to the dual mechanism, including the improvement of the enzyme stability and the antibiofilm activity of CMCS nanoparticles.