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从890中纯化得到的葡聚糖蔗糖酶与植物多酚的相互作用。

Interactions of dextransucrase purified from 890 with plant polyphenols.

作者信息

Goyal Dimple, Ahmad Shabeer, Mahmood Akhtar, Chander Sharma Sukesh

机构信息

Department of Biochemistry, Panjab University, Chandigarh, India.

出版信息

Biochem Biophys Rep. 2021 Mar 11;26:100980. doi: 10.1016/j.bbrep.2021.100980. eCollection 2021 Jul.

Abstract

Plant polyphenols have been extensively studied for their chemopreventive properties for human health. Dextransucrase plays an essential role in synthesizing exopolysaccharides from its exclusive substrate sucrose in . In the present study, the effect of polyphenols gallic acid and tannic acid was investigated on the dextransucrase activity. The enzyme was purified by ethanol precipitation followed by column chromatography by Sephadex G-200 gel chromatography, followed by PEG-400 treatment. The purified enzyme exhibited 52 fold enrichment with 17.5% yield and specific activity of 3.54 Units/mg protein. On SDS-PAGE enzyme protein gave a single band with a molecular weight of 160 kDa. Dextransucrase activity was inhibited 80-90% by 0.04 mM tannic acid (TA) or 0.4 mM gallic acid (GA) suggesting that tannic acid has 10- fold more inhibitory potential than gallic acid on the activity of dextransucrase. CD/ORD studies revealed modifications in the tertiary structure of enzyme protein in presence of tannic acid and gallic acid, which were further confirmed by fluorescence spectra of the protein in presence of tannic acid. These results suggest that inhibition of dextransucrase activity in by polyphenols may have potential applications in the prevention and control of dental caries.

摘要

植物多酚因其对人类健康的化学预防特性而受到广泛研究。葡聚糖蔗糖酶在利用其唯一底物蔗糖合成胞外多糖的过程中起着至关重要的作用。在本研究中,研究了多酚没食子酸和单宁酸对葡聚糖蔗糖酶活性的影响。通过乙醇沉淀,然后用Sephadex G - 200凝胶柱色谱法,接着进行PEG - 400处理来纯化该酶。纯化后的酶富集了52倍,产率为17.5%,比活性为3.54单位/毫克蛋白质。在SDS - PAGE上,酶蛋白呈现出一条分子量为160 kDa的单一带。0.04 mM单宁酸(TA)或0.4 mM没食子酸(GA)可抑制葡聚糖蔗糖酶活性的80 - 90%,这表明单宁酸对葡聚糖蔗糖酶活性的抑制潜力比没食子酸高10倍。圆二色光谱/旋光色散(CD/ORD)研究揭示了在单宁酸和没食子酸存在下酶蛋白三级结构的变化,单宁酸存在时蛋白质的荧光光谱进一步证实了这一点。这些结果表明,多酚对葡聚糖蔗糖酶活性的抑制作用在龋齿的预防和控制中可能具有潜在应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6dd3/7967010/d6ec1f405d2f/fx1.jpg

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