Please use this identifier to cite or link to this item: http://localhost:80/xmlui/handle/123456789/20031
Title: Isolation, Purification and Characterization of Acid Phosphatase from Germinating Vigna radiata Seeds.
Authors: SADIA NADIR
ASMA SAEED
RUBINA NAZ
AISHA SIDDIQUA
MEHRIN SHERAZI
SULTAN MEHMOOD WAZIR
AHMAD SAEED
Keywords: Acid phosphatase
Vigna radiata
purification; characterization
seedlings; isoenzyme
Issue Date: 20-Jun-2012
Publisher: HEJ Research Institute of Chemistry, University of Karachi, Karachi.
Citation: Nadir, S., Saeed, A., Naz, R., Siddiqua, A., Sherazi, M., Wazir, S. M., & Saeed, A. (2012). Isolation, purification and characterization of acid phosphatase from germinating Vigna radiata seeds. Journal of the Chemical Society of Pakistan, 34(3).
Abstract: The acid phosphatase (EC 3.1.3.2 ) has been purified from germinating seeds of vigna radiata through ammonium sulphate precipitation, DEAE-Cellulose chromatography and concanavalin A-Sepharose 4B chromatography. The specific activity of 1291 nkat.mg⁻¹of protein was obtained with recovery of nearly 1%. About 222 times purification was achieved. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) resolved two bands of acid phosphatase corresponding to molecular weight of 29 kilo Dalton (kDa) and 18 kDa. The molecular weights of native enzyme determined by gel filtration on calibrated Sephadex G-100 column were found to be 29 kDa and 18 kDa The apparent Km value of 29 kDa isoenzyme with p-nitrophenyl phosphate (pNPP) as substrate was 0.3 mM and Vmax was 1336 nmol.sec⁻¹.mg⁻¹ of protein. The optimal pH for this enzyme was 5.5 and pH stability was 4-7. It had optimum temperature of 50oC and temperature stability was 0- 50oC. The enzyme hydrolysed various phosphorylated compounds non-specifically. It was competitively inhibited by phosphate, vanadate while fluoride showed non- competitive inhibition and molybdate exhibited an inhibition of mixed type. It was found insensitive to tartrate and concluded that this enzyme was recognized as tartrate resistant acid phosphatase.
URI: http://142.54.178.187:9060/xmlui/handle/123456789/20031
ISSN: 0253-5106
Appears in Collections:Issue 03

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