期刊
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
卷 61, 期 38, 页码 9118-9124出版社
AMER CHEMICAL SOC
DOI: 10.1021/jf403044m
关键词
biogenic amines; monoamine oxidase; putrescine oxidase; bifunctional enzyme fusion
资金
- Converging Research Center Program through the National Research Foundation of Korea (NRF)
- Ministry of Education, Science and Technology [20090083290]
Biogenic amines (BAs) are a group of low-molecular-mass organic bases derived from free amino acids. Due to the undesirable effects of BAs on human health, amine oxidase-based detection methods for BAs in foods have been developed. Here, we developed a bifunctional enzyme fusion (MAPO) using a Cu2+-containing monoamine oxidase (AMAO2) and a flavin adenine dinucleotide-containing putrescine oxidase (APUO) from Arthrobacter aurescens. It was necessary to activate MARC) with supplementary Cu2+ ions, leading to a 6- to 12-fold improvement in catalytic efficiency (k(cat)/K-M) for monoamines. The optimal temperatures of Cu2+-activated MAPO (cMAPO) for both tyramine and putrescine were 50 C, and the optimal pH values for tyramine and putrescine were pH 7.0 and pH 8.0, respectively, consistent with those of AMAO2 and APUO, respectively. The cMAPO showed relative specific activities of 100, 99, 32, and 32 for 2-phenylethylamine, tyrarnine, histamine, and putrescine, respectively. The tyramine-equivalent BA contents of fermented soybean pastes by cMAPO were more than 90% of the total BA determined by HPLC. In conclusion, cMAPO is fully bifunctional toward biogenic monoamines and putrescine, allowing the combined determination of multiple BAs in foods. This colorimetric determination method could be useful for point-of-care testing to screen safety-guaranteed products prior to instrumental analyses.
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