Purification and characterization of two soluble acid invertase isozymes from Japanese pear fruit

被引:31
作者
Hashizume, H [1 ]
Tanase, K [1 ]
Shiratake, K [1 ]
Mori, H [1 ]
Yamaki, S [1 ]
机构
[1] Nagoya Univ, Grad Sch Bioagr Sci, Hort Sci Lab, Chikusa Ku, Nagoya, Aichi 4648601, Japan
关键词
Japanese pear (Pyrus serotina) fruit; soluble acid invertase (AIV; EC 3.2.1.26); enzyme purification; isozymes of AIV;
D O I
10.1016/S0031-9422(03)00107-9
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Two isozymes (AIV I and AIV II) of soluble acid invertase (EC 3.2.1.26) were purified from Japanese pear fruit through procedures including (NH4)(2)SO4 precipitating, DEAE-Sephacel column chromatography, Concanavalin A (ConA)-Sepharose affinity chromatography, hydroxyapatite column chromatography and Mono Q HR 515 column chromatography. The specific activities of purified AIV I and AIV II were 2670 and 2340 (nkat/mg protein), respectively. AIV I was a monomeric enzyme of 80 kDa, while AIV II may be also a monomeric enzyme, which is easy to be cleaved to 52 kDa and 34 kDa polypeptide during preparation by SDS-PAGE. The Km values for sucrose of AIV I and AIV II were 3.33 and 4.58 mM, respectively, and optimum pH of both enzyme activities was pH 4.5. (C) 2003 Published by Elsevier Science Ltd.
引用
收藏
页码:125 / 129
页数:5
相关论文
共 28 条
[1]  
ARAI M, 1991, PLANT CELL PHYSIOL, V32, P1291
[2]   CLONING AND SEQUENCE OF CDNAS FOR AN INTRACELLULAR ACID INVERTASE FROM ETIOLATED HYPOCOTYLS OF MUNG BEAN AND EXPRESSION OF THE GENE DURING GROWTH OF SEEDLINGS [J].
ARAI, M ;
MORI, H ;
IMASEKI, H .
PLANT AND CELL PHYSIOLOGY, 1992, 33 (03) :245-252
[3]  
BERNT E, 1974, METHOD ENZYMAT AN, V3, P1304
[4]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[5]   PURIFICATION OF SOLUBLE INVERTASE FROM POTATO [J].
BURCH, LR ;
DAVIES, HV ;
CUTHBERT, EM ;
MACHRAY, GC ;
HEDLEY, P ;
WAUGH, R .
PHYTOCHEMISTRY, 1992, 31 (06) :1901-1904
[6]   Sugar accumulation in grape berries - Cloning of two putative vacuolar invertase cDNAs and their expression in grapevine tissues [J].
Davies, C ;
Robinson, SP .
PLANT PHYSIOLOGY, 1996, 111 (01) :275-283
[7]   CHARACTERIZATION AND DISTRIBUTION OF INVERTASE ACTIVITY IN DEVELOPING MAIZE (ZEA-MAYS) KERNELS [J].
DOEHLERT, DC ;
FELKER, FC .
PHYSIOLOGIA PLANTARUM, 1987, 70 (01) :51-57
[8]   JERUSALEM ARTICHOKE INVERTASES - IMMUNOCHARACTERIZATION OF A SOLUBLE FORM AND ITS PUTATIVE PRECURSOR [J].
GOUPIL, P ;
CROISILLE, Y ;
CROISILLE, F ;
LEDOIGT, G .
PLANT SCIENCE, 1988, 54 (01) :45-54
[9]   Antisense acid invertase (TIV1) gene alters soluble sugar composition and size in transgenic tomato fruit [J].
Klann, EM ;
Hall, B ;
Bennett, AB .
PLANT PHYSIOLOGY, 1996, 112 (03) :1321-1330
[10]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+