caa a22 4500 477100597 CHVBK 20180405111540.0 cr unu---uuuuu 170330e19960101xx s 000 0 eng 10.1007/BF00197581 doi (NATIONALLICENCE)springer-10.1007/BF00197581 Purification and characterisation of soluble invertases from leaves of Arabidopsis thaliana [Elektronische Daten] [Xiuwen Tang, Hans-Peter Ruffner, Julie Scholes, Stephen Rolfe] Multiple isoforms of β-fructofuranosidase (invertase, EC 3.2.1.26) were identified in mature green leaves of the cruciferous plant Arabidopsis thaliana (L.) Heynh. There were four major and one minor isoforms of soluble acid invertase and an additional activity which could be released from the cell wall by buffers of high ionic strength. This study reports the separation and characterisation of three soluble isoforms following ammonium sulphate and polyethylene glycol 6000 precipitations, Concanavalin A, MonoQ ion exchange, Superose 12 sizeexclusion chromatography and chromatofocusing. These isoforms, designated INV1, INV2 and INV3, had isoelectric points of 4.75, 4.70 and 4.65 and a K m for sucrose of 5, 12 and 5 mM, respectively. Each had a pH optimum of 5.5, exhibited optimal activity at 45 °C and used sucrose as the preferred substrate. All fractions containing these isoforms contained a 52-kDa polypeptide which was specifically detected by immunoblotting with an antibody raised against deglycosylated wheat invertase. The N-terminal amino-acid sequence of this polypeptide was homologous to acid invertases isolated from other plant species. The possible origin of isoforms of soluble acid invertase is discussed. Springer-Verlag, 1996 Arabidopsis (acid invertase) nationallicence β-Fructofuranosidase nationallicence Enzyme purification nationallicence Invertase nationallicence PEG : polyethylene glycol nationallicence pI : isoelectric point nationallicence PMSF : phenylmethylsulphonyl fluoride nationallicence Tang Xiuwen Department of Animal and Plant Sciences, University of Sheffield, S10 2UQ, Sheffield, UK aut Ruffner Hans-Peter Institute of Plant Biology, University of Zurich, Zollikerstrasse 107, CH-8008, Zurich, Switzerland aut Scholes Julie Department of Animal and Plant Sciences, University of Sheffield, S10 2UQ, Sheffield, UK aut Rolfe Stephen Department of Animal and Plant Sciences, University of Sheffield, S10 2UQ, Sheffield, UK aut Planta Springer-Verlag 198/1(1996-01-01), 17-23 0032-0935 198:1<17 1996 198 425 https://doi.org/10.1007/BF00197581 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00197581 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Tang Xiuwen Department of Animal and Plant Sciences, University of Sheffield, S10 2UQ, Sheffield, UK aut NATIONALLICENCE 700 1- Ruffner Hans-Peter Institute of Plant Biology, University of Zurich, Zollikerstrasse 107, CH-8008, Zurich, Switzerland aut NATIONALLICENCE 700 1- Scholes Julie Department of Animal and Plant Sciences, University of Sheffield, S10 2UQ, Sheffield, UK aut NATIONALLICENCE 700 1- Rolfe Stephen Department of Animal and Plant Sciences, University of Sheffield, S10 2UQ, Sheffield, UK aut NATIONALLICENCE 773 0- Planta Springer-Verlag 198/1(1996-01-01), 17-23 0032-0935 198:1<17 1996 198 425 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer