TY - JOUR
T1 - Purification, characterization, and crystallization of monoamine oxidase from escherichia coli k-12
AU - Roh, Jung Hyeob
AU - Suzuki, Hideyuki
AU - Azakami, Hiroyuki
AU - Yamashita, Mitsuo
AU - Murooka, Yoshikatsu
AU - Kumagai, Hidehiko
N1 - Funding Information:
Acknowledgments. We wish to thank Dr. Y. Kohara, National Institute of Genetics, for providing the 5Fl phage clone. We also thank Drs. K. So~a, ~. Esaki, and. M. ~shiuchi, Institute for Chemical Research. Kyoto Umverslty, for helpmg wIth the atomic absorption alaysis.
PY - 1994/1
Y1 - 1994/1
N2 - The gene for monoamine oxidase (MAO) was cloned from an Escherichia coli genomic library and MAO was overproduced in the periplasmic space. The enzyme was purified to homogeneity by preparation of a periplasmic fraction, followed by ammonium sulfate fractionation and DEAE-cellulose column chromatography. Crystals were obtained by the hanging drop method using sodium citrate as a precipitant. The enzyme was found to be a dimer of identical subunits with a molecular weight of 80,000, and showed the highest activity at pH 7.5 and 45°C. The enzyme was inhibited by a MAO specific inhibitor, hydroxylamine, hydrazine, phenelzine, isoniazid, and tranycypromine. The enzyme oxidized tyramine, phenethylamine, and tryptamine at higher rates, but not oxidized diamine and polyamines such as putrecine and spermine. The antibody against E. coli MAO cross-reacted with purified MAO A from Klebsiella aerogenes.
AB - The gene for monoamine oxidase (MAO) was cloned from an Escherichia coli genomic library and MAO was overproduced in the periplasmic space. The enzyme was purified to homogeneity by preparation of a periplasmic fraction, followed by ammonium sulfate fractionation and DEAE-cellulose column chromatography. Crystals were obtained by the hanging drop method using sodium citrate as a precipitant. The enzyme was found to be a dimer of identical subunits with a molecular weight of 80,000, and showed the highest activity at pH 7.5 and 45°C. The enzyme was inhibited by a MAO specific inhibitor, hydroxylamine, hydrazine, phenelzine, isoniazid, and tranycypromine. The enzyme oxidized tyramine, phenethylamine, and tryptamine at higher rates, but not oxidized diamine and polyamines such as putrecine and spermine. The antibody against E. coli MAO cross-reacted with purified MAO A from Klebsiella aerogenes.
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U2 - 10.1080/bbb.58.1652
DO - 10.1080/bbb.58.1652
M3 - Article
C2 - 7765483
AN - SCOPUS:0028500866
SN - 0916-8451
VL - 58
SP - 1652
EP - 1656
JO - Bioscience, Biotechnology and Biochemistry
JF - Bioscience, Biotechnology and Biochemistry
IS - 9
ER -