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阿图罗芒果中多酚氧化酶活性的表征

Characterization of polyphenol oxidase activity in Ataulfo mango.

作者信息

Cheema Summervir, Sommerhalter Monika

机构信息

Department of Chemistry and Biochemistry, California State University East Bay, 25800 Carlos Bee Blvd., Hayward, CA 94542, USA.

Department of Chemistry and Biochemistry, California State University East Bay, 25800 Carlos Bee Blvd., Hayward, CA 94542, USA.

出版信息

Food Chem. 2015 Mar 15;171:382-7. doi: 10.1016/j.foodchem.2014.09.011. Epub 2014 Sep 16.

Abstract

Crude extracts of Ataulfo exhibited polyphenol oxidase (PPO) activity with pyrogallol, 3-methylcatechol, catechol, gallic acid, and protocatechuic acid. The substrate dependent pH optima ranged from pH 5.4 to 6.4 with Michaelis-Menten constants between 0.84 ± 0.09 and 4.6 ± 0.7 mM measured in MES or phosphate buffers. The use of acetate buffers resulted in larger Michaelis-Menten constants, up to 14.62 ± 2.03 mM. Sodium ascorbate, glutathione, and kojic acid are promising inhibitors to prevent enzymatic browning in Ataulfo. PPO activity increased with ripeness and was always higher in the skin compared to the pulp. Sodium dodecyl sulphate (SDS) enhanced PPO activity, with pulp showing a stronger increase than skin. SDS-PAGE gels stained for catecholase activity showed multiple bands, with the most prominent bands at apparent molecular weights of 53, 112, and 144 kDa.

摘要

阿图尔福芒果的粗提物对连苯三酚、3-甲基儿茶酚、儿茶酚、没食子酸和原儿茶酸表现出多酚氧化酶(PPO)活性。在MES或磷酸盐缓冲液中测得的底物依赖性pH最适范围为pH 5.4至6.4,米氏常数在0.84±0.09和4.6±0.7 mM之间。使用醋酸盐缓冲液会导致米氏常数更大,高达14.62±2.03 mM。抗坏血酸钠、谷胱甘肽和曲酸是防止阿图尔福芒果发生酶促褐变的有前景的抑制剂。PPO活性随成熟度增加,且果皮中的活性始终高于果肉。十二烷基硫酸钠(SDS)增强了PPO活性,果肉的增加幅度比果皮更大。对儿茶酚酶活性进行染色的SDS-PAGE凝胶显示出多条带,最明显的条带表观分子量为53、112和144 kDa。

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