大肠杆菌表达类黄酮O-甲基转移酶合成槲皮素甲基化衍生物
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Synthesis of Quercetin Methylated Derivatives by Expression of FOMT in Escherichia coli
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    摘要:

    槲皮素是一种重要的柑橘类黄酮,具有抗菌、抗炎和抗氧化等生物学活性,但水溶性和脂溶性较差,研究表明可通过甲基化提高其代谢稳定性和生物利用度,目前微生物转化法是获得甲基化槲皮素的良好方法。作者筛选了11种不同来源的类黄酮O-甲基转移酶(FOMT),并根据甲基化位点进行分类构建相应的大肠杆菌工程菌,以槲皮素为底物进行发酵分别合成了4种单O-甲基槲皮素(柽柳黄素、鼠李素、异鼠李素和3-O-甲基槲皮素),最高产量分别为31.17、11.17、8.90、52.95 mg/L。随后构建了大肠杆菌共培养体系,分步添加含有MpOMT4和OsNOMT的重组大肠杆菌全细胞催化剂,并通过调整体系中两种菌体的比例和生物量,最终确定菌体细胞质量浓度为24 g/L(以细胞干质量计)、两种菌体质量比为1∶2时,4′,7-二甲氧基槲皮素的最高产量为21.56 mg/L。

    Abstract:

    Quercetin is a kind of important citrus flavonoid with antibacterial, anti-inflammatory and antioxidant activities, but with poor water solubility and lipid solubility. It has been proved that the metabolic stability and bioavailability of quercetin can be improved by methylation, and microbial transformation method is a good method to obtain methylated quercetin. In this study, 11 flavonoid O-methyltransferase(FOMT) from different sources were screened and classified according to methylation sites, and then the corresponding Escherichia coli engineering bacteria were constructed. Four mono O-methylated quercetin(tamarixetin, rhamnetin, isorhamnetin and 3-O-methylquercetin)were synthesized by fermentation with quercetin as substrate. The highest yields were 31.17, 11.17, 8.90, 52.95 mg/L, respectively. Then, the co-culture system of E.coli was constructed, and the whole cell catalyst containing MpOMT4 and OsNOMT was added step by step. By adjusting the proportion and biomass of the two bacteria in the system, it was finally determined that when the dry weight of cell was 24 g/L and the ratio of cell 1 and cell 2 was 1∶2, and the highest yields of 4′, 7-dimethoxyquercetin was 21.56 mg/L.

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田苗苗,郭佳婧,刘娟,单杨.大肠杆菌表达类黄酮O-甲基转移酶合成槲皮素甲基化衍生物[J].食品与生物技术学报,2022,41(7):111-119.

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  • 在线发布日期: 2022-08-05
  • 出版日期: 2022-07-25

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