Cloning, Sequencing and Characterization of a Novel Phosphatase Gene, phoI, from Soil Bacterium Enterobacter sp. 4 |
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| Authors: | Seung Ha Kang Kwang Keun Cho Jin Duck Bok Sung Chan Kim Jaie Soon Cho Peter Chang-whan Lee Sang Kee Kang Hong Gu Lee Jung Hee Woo Hyun Jeong Lee Sang Cheol Lee Yun Jaie Choi |
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| Institution: | (1) School of Agricultural Biotechnology, Seoul National University Seoul, 151-921, Korea;(2) Department of International Livestock Industry, Chinju National University, 150 Chilamdong, Chinju, Kyongnam, 660-758, Korea;(3) R&D Center, Choong Ang Biotech Co., Ltd., 833-6 Wonsi-dong, Ansan-si, Kyunggi-do, Korea;(4) Section on Biophysical Chemistry, Laboratory of Molecular Biology, National Institute of Mental Health, 36 RM 1B08, 36 Convent Drive, Bethesda, MD 20892-4034, USA;(5) Department of Nutrition Physiology, National Livestock Research Institute, R. D. A. |
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| Abstract: | A gene, phoI, coding for a phosphatase from Enterobacter sp. 4 was cloned in Escherichia coli and sequenced. Analysis of the sequence revealed one open reading frame (ORF) that encodes a 269–amino acid protein with
a calculated molecular mass of 29 kDa. PhoI belongs to family B acid phosphatase and exhibits 49.4% identity and 62.4% homology to the hel gene from Heamophilus influenzae, which encoded an outer membrane protein (P4). The optimum pH and temperature for phosphatase activity were pH 5.5 and 40°C,
respectively. Its specific activity on ρ-nitrophenyl phosphatate was 70 U/mg at pH 5.5 and 40°C. Enzyme activity was inhibited
by Al3+, EDTA, and DTT, but fivefold activated by Cu2+ ion (350 U/mg). PhoI showed a strong synergistic effect when used with a purified E. coli phytase, AppA, to estimate combination effects.
Seung Ha Kang and Kwang Keun Cho contributed equally to this work. |
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