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Extraction of cellulose-synthesizing activity of Gluconacetobacter xylinus by alkylmaltoside
Authors:Hashimoto Akira  Shimono Kenji  Horikawa Yoshiki  Ichikawa Tsukasa  Wada Masahisa  Imai Tomoya  Sugiyama Junji
Institution:aResearch Institute of Sustainable Humanosphere (RISH), Kyoto University, Uji, Kyoto 611-0011, Japan;bGraduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-0011, Japan;cDepartment of Plant & Environmental New Resources, College of Life Sciences, Kyung Hee University, 1, Seocheon-dong, Giheung-ku, Yongin-si, Gyeonggi-do 446-701, Republic of Korea
Abstract:This study reinvestigated the synthesis of cellulose in vitro with a well-known cellulose-producing bacterium, Gluconacetobacter xylinus. Alkylmaltoside detergents, which are more frequently used in recent structural biological researches, are uniquely used in this study to solubilize cellulose-synthesizing activity from the cell membrane of G. xylinus. Activity comparable to that previously reported is obtained, while the synthesized cellulose is crystallized into a non-native polymorph of cellulose (cellulose II) as well as the previous studies. In spite of this failure to recover the native activity to synthesize cellulose I microfibril in vitro, the product is a polymer with a degree of polymerization greater than 45 as determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). It was thus concluded that the established protocol can solubilize cellulose-synthesizing activity of G. xylinus with polymerizing activity.
Keywords:Cellulose synthase  CesA/BcsA  CesB/BcsB  Cellulose I  Gluconacetobacter xylinus  Membrane protein enzyme
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