Purification and Biochemical Characterization of Insoluble Acid Invertase (INAC-INV) from Pea Seedlings SCIE SCOPUS

Cited 7 time in WEB OF SCIENCE Cited 8 time in Scopus
Title
Purification and Biochemical Characterization of Insoluble Acid Invertase (INAC-INV) from Pea Seedlings
Author(s)
Kim, Donggiun; Lee, Gunsup; Chang, Man; Park, Jongbum; Chung, Youngjae; Lee, Sukchan; Lee, Taek-Kyun
KIOST Author(s)
Lee, Taek Kyun(이택견)
Publication Year
2011-10-26
Abstract
Invertase (EC 3.2.1.26) catalyzes the hydrolysis of sucrose into D-glucose and D-fructose. Insoluble acid invertase (INAC-INV) was purified from pea (Pisum sativum L.) by sequential procedures entailing ammonium sulfate precipitation, ion exchange chromatography, absorption chromatography, reactive green-19 affinity chromatography, and gel filtration. The purified INAC-INV had a pH optimum of 4.0 and a temperature optimum of 45 degrees C. The effects of various concentrations of Tris-HCl, HgCl2, and CuSO4 on the activities of the purified invertase were examined. INAC-INV was not affected by Tris-HCl and HgCl2. INAC-INV activity was inhibited by 6.2 mM CuSO4 up to 50%. The enzymes display typical hyperbolic saturation kinetics for sucrose hydrolysis. The K-m and V-max values of INAC-INV were determined to be 4.41 mM and 8.41 U (mg protein)(-1) min(-1), respectively. INAC-INV is a true member of the beta-fructofuranosidases, which can react with sucrose and raffinose as substrates. SDS-PAGE and immunoblotting were used to determine the molecular mass of INAC-INV to be 69 kDa. The isoelectric point of INAC-INV was estimated to be about pH 8.0. Taken together, INAC-INV is a pea seedling invertase with a stable and optimum activity at lower acid pH and at higher temperature than other invertases.
ISSN
0021-8561
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/3801
DOI
10.1021/jf201057c
Bibliographic Citation
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, v.59, no.20, pp.11228 - 11233, 2011
Publisher
AMER CHEMICAL SOC
Subject
CELL-WALL INVERTASE; BETA-FRUCTOSIDASE; ALKALINE INVERTASE; SOLUBLE INVERTASES; RADISH SEEDLINGS; FRUCTOFURANOSIDASE; CARROT; LEAVES; EXPRESSION; INFECTION
Keywords
insoluble acid invertase; purification; characterization; Pisum sativum L.
Type
Article
Language
English
Document Type
Article
Publisher
AMER CHEMICAL SOC
Related Researcher
Research Interests

Marine virus,Marine pathogens,해양바이러스,해양병원체

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