African Journal of Biotechnology
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African Journal of Biotechnology Vol. 5 (1), pp. 035-044, 2 January 2006 ISSN 1684–5315 © 2006 Academic Journals
Purification and
characterization of three low-molecular-weight acid phosphatases from
peanut (Arachis hypogaea) seedlings
Jean Tia Gonnety1, Sébastien Niamké2,*,
Betty Meuwiah Faulet1, Eugène Jean-Parfait N’guessan Kouadio1
and Lucien Patrice Kouamé1.
1Laboratoire de Biochimie et
Technologie des Aliments de l’Unité de Formation et de Recherche en
Sciences et Technologie des Aliments de l’Université d’Abobo-Adjamé, 02,
BP 801 Abidjan 02, Côte d’Ivoire.
2Laboratoire de Biotechnologies,
Filière Biochimie-Microbiologie de l’Unité de Formation et de Recherche en
Biosciences de l’Université de Cocody-Abidjan, 22 BP
582 Abidjan 22, Côte d’Ivoire.
*Corresponding authors E-mail:
niamkes@yahoo.fr. Fax : (225) 20 37 81 18. Tel : (225) 07 84 64 09.
Accepted 9 November, 2005 |
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| Abstract | |||||
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The maximum acid phosphatasic activity was detected in peanut seedlings the 5th day of germination. At least, three acid phosphatases were identified and purified by successive chromatography separations on DEAE-Sepharose CL-6B, CM-Sepharose CL-6B, Sephacryl S-200 HR, and Phenyl-Sepharose HP to apparent homogeneity from developing five days old peanut seedlings. These enzymes designated acid phosphatase PI, PIIa and PIIb had native molecular weights of approximately 25.3, 22.4 and 24 kDa, respectively by gel permeation. SDS-PAGE of the purified acid phosphatase PI resolved two closely protein bands that migrated to approximately 14 and 12 kDa. Thus, this acid phosphatase likely functions as a heterodimer. Acid phosphatases PIIa and PIIb migrated as single band (each) with a similar molecular weight estimated to 21 kDa. The three enzymes had a similar optima pH (5.0) and temperature (55°C), and appeared to be stable in the presence of non-ionic detergents such as Triton X-100, Nonidet P 40 as well as Na+ and K+. Substrate specificity indicated that the three acid phosphatases hydrolyzed a broad range of phosphorylated substrates. However, natural substrates such as ADP and ATP were the compounds with highest rate of hydrolysis for acid phosphatase PI, while acid phosphatase PIIa exhibited phytasic activity. These results indicate that each purified acid phosphatase from peanut seedlings played a peculiar role during germination.
Key words: acid phosphatase; seedling; peanut; arachis hypogaea; germination; low-molecular-weight.
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