Thermo-Magnetoresponsive Dual Function Nanoparticles: An Approach for Magnetic Entrapable–Releasable Chitosan

Magnetic polymeric nanoparticles can be used for selective binding in a magnetic field. However, as the magnetic nanoparticles (MAG) are stabilized with polymers, the separation of the MAG from the polymer chains after use is difficult. This work proposes a combination of a thermoresponsive polymer with MAG allows for the as-desired simple removal of MAG from the polymer chains. For this, chitosan (CS) was conjugated with thermoresponsive poly­(<i>N</i>-isopropylacrylamide) (PNIPAM) and antibody (Ab) together with the physisorbed MAG as a thermo-magneto dual functional material. The key synthesis steps are (i) radical polymerization of NIPAM in the presence of mercaptoacetic acid so that the PNIPAM obtained contains terminal carboxylic acid groups (PNIPAM-COOH), (ii) the CS-<i>N</i>-hydroxysuccinamide water-based system that allows conjugation of CS with PNIPAM-COOH in water at room temperature, and (iii) the weak interaction between MAG and the CS chain. As a model application, CS is conjugated with the antirecombinant <i>Leptospirosis</i> Ab (rLipL32) to allow the selective binding and collection of the target antigen under the dual functions. This is the first demonstration of a simple but effective solution for MAG exclusion from the target molecules and will be practical for diverse applications, such as diagnosis, sensors, filtration, etc

Modified properties of alternan polymers arising from deletion of SH3-like motifs in Leuconostoc citreum ABK-1 alternansucrase

Alternansucrase (ALT, EC 2.4.1.140) catalyses the formation of an alternating 〈-1, 3/1, 6-linked glucan, with periodic branch points, from sucrose substrate. Beyond the catalytic domain, this enzyme harbours seven additional C-terminal SH3-like repeats. We herein generated two truncated alternansucrases, possessing deletions of three and seven adjacent SH3 motifs, giving Δ3SHALT and Δ7SHALT. Δ3SHALT and Δ7SHALT exhibited kcat/Km for transglycosylation activity 2.3- and 1.5-fold lower than wild-type ALT (WTALT), while hydrolysis was detected only in the truncated ALTs, oligosaccharide patterns and polymer glycosidic linkage were similar to that of WTALT. The viscosities of ALT polymers increase by ˜100-fold at 15% (w/v), with gel-like states formed at 12.5, 15.0, and 20.0% (w/v) produced by polymer from WTALT, Δ3SHALT, and Δ7SHALT, respectively. The average nanoparticle sizes of Δ3SHALT and Δ7SHALT polymers were 80 nm, compared to 90 nm from WTALT. In conclusion, even relatively subtle differences in the structure of ALT-produced alternan give rise to profound impact on the glucan polymer physicochemical properties