caa a22 4500 605500347 CHVBK 20210128100555.0 cr unu---uuuuu 210128e20150401xx s 000 0 eng 10.1007/s00253-014-6116-6 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6116-6 Multifunctional cellulolytic auxiliary activity protein Hc AA10-2 from Hahella chejuensis enhances enzymatic hydrolysis of crystalline cellulose [Elektronische Daten] [Sunil Ghatge, Amar Telke, Tatoba Waghmode, Yuno Lee, Keun-Woo Lee, Doo-Byoung Oh, Hyun-Dong Shin, Seon-Won Kim] The modular auxiliary activity (AA) family of proteins is believed to cause amorphogenesis in addition to oxidative cleavage of crystalline cellulose although the supporting evidence is limited. HcAA10-2 is a modular AA10 family protein (58kDa) composed of a AA10 module and a family two carbohydrate binding module (CBM2), joined by a long stretch of 222 amino acids of unknown function. The protein was expressed in Escherichia coli and purified to homogeneity. Scanning electron microscopy and X-ray diffraction analysis of Avicel treated with HcAA10-2 provided evidence for the disruption of the cellulose microfibrils ("amorphogenesis”) and reduction of the crystallinity index, resulting in a twofold increase of cellulase adsorption on the polysaccharide surface. HcAA10-2 exhibited weak endoglucanase-like activity toward soluble cellulose and cello-oligosaccharides with an optimum at pH 6.5 and 45°C. HcAA10-2 catalyzed oxidative cleavage of crystalline cellulose released native and oxidized cello-oligosaccharides in the presence of copper and an electron donor such as ascorbic acid. Multiple sequence alignment indicated that His1, His109, and Phe197 in the AA10 module formed the conserved copper-binding site. The reducing sugar released from Avicel by the endoglucanase Cel5 and Celluclast accompanying HcAA10-2 was increased by four- and sixfold, respectively. Moreover, HcAA10-2 and Celluclast acted synergistically on pretreated wheat straw biomass resulting in a threefold increase in reducing sugar than Celluclast alone. Taken together, these results suggest that HcAA10-2 is a novel multifunctional modular AA10 protein possessing amorphogenesis, weak endoglucanase, and oxidative cleavage activities useful for efficient degradation of crystalline cellulose. Springer-Verlag Berlin Heidelberg, 2014 Lytic polysaccharide monooxygenases (LPMOs) nationallicence AA10 (formerly CBM33) nationallicence Amorphogenesis nationallicence Endoglucanase nationallicence Oxidative cleavage nationallicence Ghatge Sunil Division of Applied Life Sciences (BK21), SSAC and PMBBRC, Gyeongsang National University, 660-701, Jinju, Republic of Korea aut Telke Amar Division of Applied Life Sciences (BK21), SSAC and PMBBRC, Gyeongsang National University, 660-701, Jinju, Republic of Korea aut Waghmode Tatoba Division of Applied Life Sciences (BK21), SSAC and PMBBRC, Gyeongsang National University, 660-701, Jinju, Republic of Korea aut Lee Yuno Division of Applied Life Sciences (BK21), SSAC and PMBBRC, Gyeongsang National University, 660-701, Jinju, Republic of Korea aut Lee Keun-Woo Division of Applied Life Sciences (BK21), SSAC and PMBBRC, Gyeongsang National University, 660-701, Jinju, Republic of Korea aut Oh Doo-Byoung Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Republic of Korea aut Shin Hyun-Dong School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, 30332-0100, Atlanta, GA, USA aut Kim Seon-Won Division of Applied Life Sciences (BK21), SSAC and PMBBRC, Gyeongsang National University, 660-701, Jinju, Republic of Korea aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/7(2015-04-01), 3041-3055 0175-7598 99:7<3041 2015 99 253 https://doi.org/10.1007/s00253-014-6116-6 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6116-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ghatge Sunil Division of Applied Life Sciences (BK21), SSAC and PMBBRC, Gyeongsang National University, 660-701, Jinju, Republic of Korea aut NATIONALLICENCE 700 1- Telke Amar Division of Applied Life Sciences (BK21), SSAC and PMBBRC, Gyeongsang National University, 660-701, Jinju, Republic of Korea aut NATIONALLICENCE 700 1- Waghmode Tatoba Division of Applied Life Sciences (BK21), SSAC and PMBBRC, Gyeongsang National University, 660-701, Jinju, Republic of Korea aut NATIONALLICENCE 700 1- Lee Yuno Division of Applied Life Sciences (BK21), SSAC and PMBBRC, Gyeongsang National University, 660-701, Jinju, Republic of Korea aut NATIONALLICENCE 700 1- Lee Keun-Woo Division of Applied Life Sciences (BK21), SSAC and PMBBRC, Gyeongsang National University, 660-701, Jinju, Republic of Korea aut NATIONALLICENCE 700 1- Oh Doo-Byoung Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Republic of Korea aut NATIONALLICENCE 700 1- Shin Hyun-Dong School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, 30332-0100, Atlanta, GA, USA aut NATIONALLICENCE 700 1- Kim Seon-Won Division of Applied Life Sciences (BK21), SSAC and PMBBRC, Gyeongsang National University, 660-701, Jinju, Republic of Korea aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/7(2015-04-01), 3041-3055 0175-7598 99:7<3041 2015 99 253