Han Ruizhi, Ge Binbin, Jiang Mingyang, Xu Guochao, Dong Jinjun, Ni Ye
Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, China.
Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, China.
J Ind Microbiol Biotechnol. 2017 Sep;44(9):1343-1354. doi: 10.1007/s10295-017-1960-x. Epub 2017 Jun 28.
Genistein has been regarded as one important soy isoflavone with multiple health benefits, whereas its applications are limited by the low hydrophilicity. To improve the water solubility, codon optimized cyclodextrin glycosyltransferase from Paenibacillus macerans was employed for genistein transglycosylation in this study. At least four transglycosylation products were produced and identified by HPLC and LC-MS: genistein monoglucoside, diglucoside, triglucoside, and tetraglucoside derivatives. Obviously, the yields of genistein monoglucoside and genistein diglucoside exhibited great superiority compared with other two products. To maximize the yield of genistein diglucoside, various reaction conditions such as genistein dissolvents, glycosyl donors, substrates concentrations and ratios, enzyme concentrations, reaction pH, temperature, and time were optimized. Finally, the yield of genistein diglucoside was enhanced by 1.5-fold under the optimum reaction system. Our study demonstrates that the production of genistein diglucoside could be specifically enhanced, which is one important genistein derivative with better water solubility and stability.
染料木黄酮被认为是一种具有多种健康益处的重要大豆异黄酮，但其应用受到低亲水性的限制。为了提高其水溶性，本研究采用密码子优化的来自浸麻芽孢杆菌的环糊精糖基转移酶对染料木黄酮进行转糖基化反应。通过高效液相色谱（HPLC）和液相色谱 - 质谱联用（LC - MS）至少鉴定出四种转糖基化产物：染料木黄酮单葡萄糖苷、二葡萄糖苷、三葡萄糖苷和四葡萄糖苷衍生物。显然，染料木黄酮单葡萄糖苷和染料木黄酮二葡萄糖苷的产量与其他两种产物相比具有很大优势。为了使染料木黄酮二葡萄糖苷的产量最大化，对染料木黄酮的溶剂、糖基供体、底物浓度和比例、酶浓度、反应pH、温度和时间等各种反应条件进行了优化。最终，在最佳反应体系下，染料木黄酮二葡萄糖苷的产量提高了1.5倍。我们的研究表明，染料木黄酮二葡萄糖苷的产量可以特异性提高，它是一种具有更好水溶性和稳定性的重要染料木黄酮衍生物。
