title: Enhancing the stability and antibiofilm activity of DspB by immobilization on carboxymethyl chitosan nanoparticles.
abstract: 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 0.3 mg/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.
authors: Key Laboratory of Marine Drugs, Chinese Ministry of Education, Qingdao, China; Shandong Provincial Key Laboratory of Glycoscience and Glycotechnology, Qingdao, China; School of Medicine and Pharmacy, Ocean University of China, Qingdao, China.
authors: Yulong
authors: Y
authors: Tan
authors: Key Laboratory of Marine Drugs, Chinese Ministry of Education, Qingdao, China; Shandong Provincial Key Laboratory of Glycoscience and Glycotechnology, Qingdao, China; School of Medicine and Pharmacy, Ocean University of China, Qingdao, China.
authors: Su
authors: S
authors: Ma
authors: College of Marine Life Sciences, Ocean University of China, Qingdao, China.
authors: Chenguang
authors: C
authors: Liu
authors: Key Laboratory of Marine Drugs, Chinese Ministry of Education, Qingdao, China; Shandong Provincial Key Laboratory of Glycoscience and Glycotechnology, Qingdao, China; School of Medicine and Pharmacy, Ocean University of China, Qingdao, China.
authors: Wengong
authors: W
authors: Yu
authors: Key Laboratory of Marine Drugs, Chinese Ministry of Education, Qingdao, China; Shandong Provincial Key Laboratory of Glycoscience and Glycotechnology, Qingdao, China; School of Medicine and Pharmacy, Ocean University of China, Qingdao, China. Electronic address: fhan@ouc.edu.cn.
authors: Feng
authors: F
authors: Han
mesh_terms: D000118:Acetylglucosaminidase / Q000737:chemistry / Q000378:metabolism*; D016976:Aggregatibacter actinomycetemcomitans / Q000201:enzymology*; D000890:Anti-Infective Agents / Q000737:chemistry / Q000378:metabolism*; D018441:Biofilms / Q000187:drug effects*; D048271:Chitosan / Q000031:analogs & derivatives* / Q000378:metabolism; D003001:Cloning, Molecular; D004356:Drug Storage; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D053758:Nanoparticles / Q000378:metabolism*; D013211:Staphylococcus aureus / Q000187:drug effects / Q000502:physiology; D013212:Staphylococcus epidermidis / Q000187:drug effects / Q000502:physiology; D013696:Temperature
chemical_list: D000890:Anti-Infective Agents; D004800:Enzymes, Immobilized; C514968:carboxymethyl-chitosan; D048271:Chitosan; D000118:Acetylglucosaminidase
keywords: Biofilm; Carboxymethyl chitosan; Immobilization; Nanoparticles; Stability