caa a22 4500 477100120 CHVBK 20180405111538.0 cr unu---uuuuu 170330e19961101xx s 000 0 eng 10.1007/BF00200295 doi (NATIONALLICENCE)springer-10.1007/BF00200295 Characterization of seven basic endochitinases isolated from cell cultures of Citrus sinensis (L.) [Elektronische Daten] [Richard Mayer, T. McCollum, R. Niedz, C. Hearn, Roy McDonald, E. Berdis, H. Doostdar] Seven endochitinases (EC 3.2.1.14) (relative molecular masses 23000-28000 and isoelectric points 10.3-10.4) were purified from nonembryogenic Citrus sinensis L. Osbeck cv. Valencia callus tissue. The basic chitinase/lysozyme from this tissue (BCLVC) exhibited lysozyme, chitinase and chitosanase activities and was determined to be a class III chitinase. While BCLVC acted as a lysozyme at pH 4.5 and low ionic strength (0.03) it acted as a chitinase/chitosanase at high ionic strengths (0.2) with a pH optimum of ca. 5. The lysozyme activity of BCLVC was inhibited by histamine, imidazole, histidine and the N-acetyl-d-glucosamine oligosaccharide (GlcNAc)3. The basic chitinase from cv. Valencia callus, BCVC-2, had an N-terminal amino acid sequence similar to tomato and tobacco AP24 proteins. The sequences of the other five chitinases were N-terminal blocked. Whereas BCLVC was capable of hydrolyzing 13.8-100% acetylated chitosans and (GlcNAc)4-6 oligosaccharides, BCVC-2 hydrolyzed only 100% acetylated chitosan, and the remaining enzymes expressed varying degrees of hydrolytic capabilities. Experiments with (GlcNAc)2-6 suggest that BCLVC hydrolysis occurs in largely tetrasaccharide units whereas hydrolysis by the other chitinases occurs in disaccharide units. Cross-reactivities of the purified proteins with antibodies for a potato leaf chitinase (AbPLC), BCLVC, BCVC-3, and tomato AP24 indicate that these are separate and distinct proteins. Mention of a trademark, warranty, propriety, or vendor does not constitute a guarantee by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other products or vendors that may also be suitable. Springer-Verlag, 1996 Citrus nationallicence Chitinases nationallicence Chitosanase nationallicence Lysozymes nationallicence Pathogenesis-related proteins nationallicence Plant defense nationallicence Ab : antibody nationallicence BCLVC : basic chitinase/lysozyme cv. Valencia callus nationallicence BCVC : basic chitinase cv. Valencia callus nationallicence CE : capillary electrophoresis nationallicence CM-chitin-RBV : carboxymethyl-chitin-remazol brilliant violet nationallicence GlcNAc : N-acetyl-d-glucosamine nationallicence HEWL : hen egg-white lysozyme nationallicence Mr : relativemolecular mass nationallicence pI : isoelectric point nationallicence PLC : potato leaf chitinase nationallicence PR : pathogenesis-related nationallicence SEC : size exclusion chromatography nationallicence Mayer Richard U.S. Department of Agriculture, Agricultural Research Service, U.S. Horticultural Research Laboratory, 2120 Camden Road, FL 32803-1419, Orlando, USA aut McCollum T. U.S. Department of Agriculture, Agricultural Research Service, U.S. Horticultural Research Laboratory, 2120 Camden Road, FL 32803-1419, Orlando, USA aut Niedz R. U.S. Department of Agriculture, Agricultural Research Service, U.S. Horticultural Research Laboratory, 2120 Camden Road, FL 32803-1419, Orlando, USA aut Hearn C. U.S. Department of Agriculture, Agricultural Research Service, U.S. Horticultural Research Laboratory, 2120 Camden Road, FL 32803-1419, Orlando, USA aut McDonald Roy U.S. Department of Agriculture, Agricultural Research Service, U.S. Horticultural Research Laboratory, 2120 Camden Road, FL 32803-1419, Orlando, USA aut Berdis E. U.S. Department of Agriculture, Agricultural Research Service, U.S. Horticultural Research Laboratory, 2120 Camden Road, FL 32803-1419, Orlando, USA aut Doostdar H. U.S. Department of Agriculture, Agricultural Research Service, U.S. Horticultural Research Laboratory, 2120 Camden Road, FL 32803-1419, Orlando, USA aut Planta Springer-Verlag 200/3(1996-11-01), 289-295 0032-0935 200:3<289 1996 200 425 https://doi.org/10.1007/BF00200295 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00200295 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Mayer Richard U.S. Department of Agriculture, Agricultural Research Service, U.S. Horticultural Research Laboratory, 2120 Camden Road, FL 32803-1419, Orlando, USA aut NATIONALLICENCE 700 1- McCollum T. U.S. Department of Agriculture, Agricultural Research Service, U.S. Horticultural Research Laboratory, 2120 Camden Road, FL 32803-1419, Orlando, USA aut NATIONALLICENCE 700 1- Niedz R. U.S. Department of Agriculture, Agricultural Research Service, U.S. Horticultural Research Laboratory, 2120 Camden Road, FL 32803-1419, Orlando, USA aut NATIONALLICENCE 700 1- Hearn C. U.S. Department of Agriculture, Agricultural Research Service, U.S. Horticultural Research Laboratory, 2120 Camden Road, FL 32803-1419, Orlando, USA aut NATIONALLICENCE 700 1- McDonald Roy U.S. Department of Agriculture, Agricultural Research Service, U.S. Horticultural Research Laboratory, 2120 Camden Road, FL 32803-1419, Orlando, USA aut NATIONALLICENCE 700 1- Berdis E. U.S. Department of Agriculture, Agricultural Research Service, U.S. Horticultural Research Laboratory, 2120 Camden Road, FL 32803-1419, Orlando, USA aut NATIONALLICENCE 700 1- Doostdar H. U.S. Department of Agriculture, Agricultural Research Service, U.S. Horticultural Research Laboratory, 2120 Camden Road, FL 32803-1419, Orlando, USA aut NATIONALLICENCE 773 0- Planta Springer-Verlag 200/3(1996-11-01), 289-295 0032-0935 200:3<289 1996 200 425 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer