Purification and Biochemical Characterization of Insoluble Acid Invertase (INAC-INV) from Pea Seedlings
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Title
- Purification and Biochemical Characterization of Insoluble Acid Invertase (INAC-INV) from Pea Seedlings
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Author(s)
- Kim, Donggiun; Lee, Gunsup; Chang, Man; Park, Jongbum; Chung, Youngjae; Lee, Sukchan; Lee, Taek-Kyun
- KIOST Author(s)
- Lee, Taek Kyun(이택견)
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Alternative Author(s)
- 이건섭; 장만; 이택견
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Publication Year
- 2011-10-26
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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.
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ISSN
- 0021-8561
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URI
- https://sciwatch.kiost.ac.kr/handle/2020.kiost/3801
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DOI
- 10.1021/jf201057c
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Bibliographic Citation
- JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, v.59, no.20, pp.11228 - 11233, 2011
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Publisher
- AMER CHEMICAL SOC
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Subject
- CELL-WALL INVERTASE; BETA-FRUCTOSIDASE; ALKALINE INVERTASE; SOLUBLE INVERTASES; RADISH SEEDLINGS; FRUCTOFURANOSIDASE; CARROT; LEAVES; EXPRESSION; INFECTION
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Keywords
- insoluble acid invertase; purification; characterization; Pisum sativum L.
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Type
- Article
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Language
- English
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Document Type
- Article
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