The network structure of high acyl gellan polysaccharide was investigated using dynamic viscoelasticity and steady flow viscosity measurements, as well as atomic force microscopy (AFM). Time-temperature superposition (TTS) of mechanical spectra of aqueous dispersions having a gellan concentration of 0.1% w/w revealed a gel-like response at the lower end of the frequency range. The TTS master curve of the steady flow data exhibited a power-law relationship between shear viscosity and shear rate at the lower end of the shear rate range, instead of a Newtonian plateau. These rheological characteristics suggest the existence of an effective yield stress arising from the presence of a percolated network. AFM images of high acyl gellan revealed micrometer-sized networks composed of double-stranded helices laterally associated to varying degrees. These associated helices did not dissociate fully on heating at 90℃, suggesting that they are partially preserved native networks secreted by gellan-producing bacteria.Keywords: atomic force microscopy, high acyl gellan, network, time-temperature superposition, viscoelasticity, yield stress *To whom correspondence should be addressed. E-mail: sikeda2@wisc.edu (S.I.), qzhong@utk.edu (Q.Z.) †
IntroductionGellan is an extracellular polysaccharide secreted by Sphingomonas elodea in the form of hydrated networks encapsulating the bacterial cell (Morris, 1998). Due to its ability to form clear and heat-resistant hydrogels, gellan is widely utilized in industry (Valli and Miskiel, 2001). Naturally occurring gellan exists in the high acyl form with a tetrasaccharide repeating unit consisting of →3)-β-d-glucose-(1→4)-β-d-glucuronic acid-(1→4)-β-d-glucose-(1→4)-α-lrhamnose-(1→ (Jansson et al., 1983;O'Neil et al., 1983). On the (1→3) linked β-d-glucose residues, each C-2 position is esterified with l-glycerate, while approximately one-half of the C-6 positions are esterified with acetate (Kuo et al., 1986). High acyl gellan is alkali-treated to obtain a deacylated product for expanded commercial applications (Baird et al., 1992;Valli and Miskiel, 2001). Deacylated gellan molecules are fully hydrated in an aqueous solution at sufficiently high temperatures. Upon cooling to a certain temperature range, fully hydrated molecules in the disordered coiled conformation transform into the double-stranded helical conformation (Morris, 1998;Ikeda et al., 2004;Rinaudo, 2004). Double-stranded helices aggregate laterally to form thicker rod-like structures in the presence of specific cations, such as potassium, that shield electrostatic repulsion between double-stranded helices arising from negatively charged carboxyl groups, forming macroscopic gels at sufficiently high polysaccharide concentrations (Morris, 1998;Ikeda et al., 2004;Rinaudo, 2004). The aggregation of double-stranded helices results in a much higher gel-melting temperature than the temperature corresponding to gel formation (Annaka et al., 1999; Matsunaka et al., 1999;Miyoshi and Nishinari, 1999).High acyl gellan recovered directly...