Coumestrol (CMS), one of the soybean isoflavonoids which contains several benefits for maintaining skin function including antiageing properties. In this study, we evaluated various explant sources and plant genotypes to determine competent soybean adventitious root materials for the mass production of CMS, and investigated their skin care efficacies to be used as a novel cosmetic ingredient. Adventitious roots were directly induced from in vitro seedling derived from the mature seeds, extracts were prepared and refluxed for enzymatic deglycosylation. In vitro cell cytotoxicity was evaluated using normal human dermal fibroblast and murine B16 melanoma cells after treatment with increasing concentrations of methanol soybean adventitious roots extracts for 72 h. Finally, in vitro cell assays on HDF cells were performed to evaluate the effect of the soybean adventitious roots extracts in collagen production. The root induction frequency and biomass productivity were significantly affected by plant genotypes, explant sources, the type of auxin used and its concentration. The total CMS production (per 1 L medium) after 4 weeks of culture in a bulb-type bubble bioreactor (3 L capacity) was the highest in the adventitious roots induced from the radicles of Glycine max, ‘Sinhwakong’. Different strengths of Murashige and Skoog (MS) medium were tested to develop culture protocols and the highest total CMS production (per 1 L medium) was observed at 1/2 MS. The content of coumestrin, the glycoside form of CMS, was higher than that of CMS in the roots cultured in 1/2 MS medium for 4 weeks in a bioreactor. The final content of CMS in the ethanol extract after enzymatic deglycosylation was 81.3-fold higher than non-enzymatic deglycosylation. Almost all the coumestrin in the roots were converted to CMS. Further, the enriched CMS root did not exhibit any cell cytotoxicity in normal human dermal fibroblast (HDF) and murine B16 melanoma cells (B16) for 72 h. In addition, in vitro collagen production assay on HDF cells showed that the enriched CMS root increased the collagen production compared to the coumestrol, daidzein, and non-enzyme-treated sample. Thus, enriched CMS root could be potential ingredient for the cosmetic applications

中文翻译：

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通过生物反应器从大豆（Glycine max）不定根中大量生产香豆甾醇：对胶原蛋白生产的影响

香豆酚（CMS），一种大豆异黄酮类化合物，具有多种皮肤功效，包括抗衰老特性。在这项研究中，我们评估了各种外植体来源和植物基因型，以确定可大量生产CMS的大豆不定根材料，并研究了它们的皮肤护理功效，可用作新型化妆品成分。不定根直接从成熟种子衍生的体外幼苗中诱导，制备提取物并回流以进行酶促去糖基化。在用浓度不断增加的甲醇大豆不定根提取物处理72小时后，使用正常人皮肤成纤维细胞和鼠B16黑色素瘤细胞评估了体外细胞毒性。最后，对HDF细胞进行了体外细胞测定，以评估大豆不定根提取物在胶原蛋白生产中的作用。根的诱导频率和生物量生产力受到植物基因型，外植体来源，生长素类型及其浓度的显着影响。在鳞茎型气泡生物反应器中培养4周（容量为3 L）后，CMS的总产量（每1 L培养基）最高，这是由不定根产生的。Glycine max，“ Sinhwakong”。测试了不同强度的Murashige和Skoog（MS）培养基以开发培养方案，在1/2 MS处观察到最高的CMS总产量（每1 L培养基）。在生物反应器中于1/2 MS培养基中培养4周的根中，香豆素的含量（CMS的糖苷形式）高于CMS的含量。酶促去糖基化后，乙醇提取物中CMS的最终含量比非酶促去糖基化高81.3倍。根中几乎所有的香豆素都转化为CMS。此外，富集的CMS根在正常人真皮成纤维细胞（HDF）和鼠类B16黑色素瘤细胞（B16）中连续72 h均未表现出任何细胞毒性。此外，对HDF细胞的体外胶原蛋白生产测定表明与香豆酚，大豆黄酮和未经酶处理的样品相比，富集的CMS根增加了胶原蛋白的产生。因此，富集的CMS根可能是化妆品应用中的潜在成分