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| Effect of Phytohormones and Group Selective Reagents on Acid Phosphatase from Cladosporium cladosporioides |
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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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