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Asian Journal of Biotechnology
  Year: 2009 | Volume: 1 | Issue: 1 | Page No.: 1-11
DOI: 10.3923/ajbkr.2009.1.11
 
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Effect of Phytohormones and Group Selective Reagents on Acid Phosphatase from Cladosporium cladosporioides
Hamed M. El-Shora and M. Metwally

Abstract:
Acid phosphatase [EC 3.1.3.2] was isolated and characterized from Cldosporium cladosporioides. The activity was determined by using p-nitrophenyl phosphate (PNPP) as substrate. Gibberellic acid (GA3), 6-benzylaminopurine (BAP), kinetin and 2,4-dichlorophenoxyacetic acid (2,4 D) induced the enzyme activity when included in the growth medium. GA3 and BAP were the strongest inducers. However, indole acetic acid (IAA) did not show any effect on the enzyme activity. The effect of calmodulin antagonists on GA3- BAP-induced acid phosphatase synthesis was also investigated. The calmodulin antagonists chlorpromazine, haloperidol, trifluoroperazine and quinacrine inhibited both GA3- and BAP-induced synthesis of acid phosphatase. This leads to the suggestion that some calmodulin-controlled mechanism is involved in GA3- and BAP-induced acid phosphatase synthesis. The enzyme was purified to homogeneity on the basis of polyacrylamide gel electrophoresis using ammonium sulfate (35-80 %), Sepharcryl S-200HR and Phenyl-Sepharose HP. The final specific activity was 203.8 U mg-1 with purification fold of 328.6. The divalent cations Ba2+, Ca2+ and Sr2+ and Co2+ were strong activators whereas Zn+2 was a strong inhibitor. Ca2+ is required for activity and thermal stability of acid phosphatase. Citrate, borate and carbonate enhanced acid phosphatase. Bromide, arsenate, phosphate, sulfite, sulfate, fluoride, EDTA and EGTA inhibited the enzyme activity. N-bromosuccinimide (NBS), tetranitromethane (TNM), N-ethylmaleimide (NEM) and diethylpyrocarbonate (DEPC) inhibited acid phosphatase activity suggesting that tryptophenyl, cysteinyl and tyrosyl and histidyl residues taking part in the catalytic activity of acid phosphatase. Dithiothreitol (DTT), reduced glutathione (GHS), L-ascorbic acid and cysteine at 5 mM enhanced the enzyme activity. Triton X-100, Nonidet F40, Brij-35 and sodium oleate enhanced the acid phosphatase activity whereas sodium lauryl sulphate was inhibitor.
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How to cite this article:

Hamed M. El-Shora and M. Metwally, 2009. Effect of Phytohormones and Group Selective Reagents on Acid Phosphatase from Cladosporium cladosporioides. Asian Journal of Biotechnology, 1: 1-11.

DOI: 10.3923/ajbkr.2009.1.11

URL: https://scialert.net/abstract/?doi=ajbkr.2009.1.11

 
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