Aqueous size-exclusion chromatographic method for the quantification of cyanobacterial native glycogen

作者：Izumi Yoshihiro; Aikawa Shimpei; Matsuda Fumio; Hasunuma Tomohisa; Kondo Akihiko^*

来源：Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, 2013, 930: 90-97.

DOI：10.1016/j.jchromb.2013.04.037

• Size-exclusion chromatography
• Native glycogen
• Quantitative analysis
• Polyhydroxy methacrylate
• Microalgae
• Arthrospira platensis

Cyanobacterial glycogen has gained interest as a valuable biomass feedstock for biofuel production. However, an ideal method for native glycogen quantification has not been developed. Here, we have proposed a simple methodology that enables the quantitative determination of cyanobacterial glycogen concentration with high repeatability using aqueous size-exclusion chromatography with a differential refractive index detector (SEC/RID). Our SEC/RID system also allows size distributions for native glycogen based on hydrodynamic volumes (V-h), which is proportional to the product of the molecular mass (M) and intrinsic viscosity [eta], obtained by universal calibration using linear homopolymers of known M with Mark-Houwink-Sakurada parameters. The universal calibration curve achieved a broad linear range (V-h parameter [eta]M = 2 x 10(2)-8 x 10(8) mLg(-1)) with a high correlation coefficient (R-2 = 0.9942), because the developed system is equipped with an OHpak SB-806M HQ aqueous column containing four types of polyhydroxy methacrylate-based particles with different particle and pore sizes. Based on the SEC/RID system, response of molecular size distribution of glycogen in microalgae to the cultivation condition was first observed. Our established SEC/RID method has several advantages over conventional techniques, including the simultaneous quantitative and size distribution analyses of glycogen, and represents a potentially useful tool to elucidate the relationship between structural properties and the roles of glycogen in metabolism.

• 出版日期2013-7-1