Dwibedi Vagish, Rath Santosh Kumar, Prakash Ranjana, Saxena Sanjai
Department of Biotechnology, Thapar Institute of Engineering and Technology, Patiala, Punjab, 147001, India.
School of Chemistry and Biochemistry, G-Block, Thapar Institute of Engineering and Technology, Patiala, Punjab, 147004, India.
Biotechnol Lett. 2021 Mar;43(3):627-644. doi: 10.1007/s10529-020-03032-7. Epub 2020 Nov 6.
The present investigation primarily focusses on enhancement of resveratrol production by endophytic production from the endophytic fungus, Arcopilus aureus via one variable at a time approach (OVAT) followed by statistical approach using response surface methodology (RSM). The paper also highlights the characterization of fungal resveratrol using spectroscopic techniques. Further the tyrosinase inhibitory property was also explored in this communication for its possible use as a cosmeceutical ingredient.
Optimization of physicochemical and nutritional parameters using OVAT approach exhibited 1.23-fold enhancement in production of resveratrol when compared to initial yield, 89.1 ± 0.08 µg/mL. Further RSM resulted in 1.49-fold enhancement in resveratrol production i.e. 133.53 µg/ml. Further, 25 mg of fungal resveratrol in pure form was obtained from the spent broth of Arcopilus aureus by column chromatography using a mobile phase comprising of MeOH: DCM in a ratio of 1.75:98.25. Further its purity on TLC was checked using 5% MeOH: DCM as mobile phase. Symmetrical peak with R of 3.36 min using a C18 reverse phase column confirmed the homogeneity of the purified fungal resveratrol with standard resveratrol and further corroborated by 1H-NMR, 13C-NMR and HR-MS analysis. Fungal resveratrol exhibited a good tyrosinase inhibition with an IC of 2.654 ± 0.432 µg/mL as compared to Kojic acid (1.329 ± 0.333).
The present study has provided sufficient lead that process optimization techniques can complement each other for optimized production of bioactive compounds by microorganisms apart from strain improvement techniques which are generally adopted in the industry. The enhancement of resveratrol production by Arcopilus aureus by process optimization further opens up avenues for strain improvement for commercial resveratrol production through fermentation for nutraceutical and cosmeceutical applications.
本研究主要聚焦于通过一次改变一个变量的方法（OVAT），从内生真菌金黄色弧顶菌中内生生产白藜芦醇，随后使用响应面法（RSM）进行统计分析，以提高白藜芦醇的产量。本文还着重介绍了使用光谱技术对白藜芦醇进行真菌特性表征。此外，本通讯还探讨了其酪氨酸酶抑制特性，以评估其作为化妆品成分的潜在用途。
采用OVAT方法优化理化和营养参数后，白藜芦醇产量较初始产量89.1±0.08μg/mL提高了1.23倍。进一步通过RSM，白藜芦醇产量提高了1.49倍，即133.53μg/ml。此外，使用甲醇：二氯甲烷比例为1.75:98.25的流动相，通过柱色谱法从金黄色弧顶菌的发酵液中获得了25mg纯形式的真菌白藜芦醇。进一步使用5%甲醇：二氯甲烷作为流动相，在薄层色谱上检查其纯度。使用C18反相柱，保留时间为3.36分钟的对称峰证实了纯化的真菌白藜芦醇与标准白藜芦醇的同质性，并通过1H-NMR、13C-NMR和HR-MS分析得到进一步证实。与曲酸（1.329±0.333）相比，真菌白藜芦醇表现出良好的酪氨酸酶抑制活性，IC50为2.654±0.432μg/mL。
本研究提供了充分的线索，表明除了工业上普遍采用的菌株改良技术外，工艺优化技术可以相互补充，以实现微生物对生物活性化合物的优化生产。通过工艺优化提高金黄色弧顶菌生产白藜芦醇的产量，进一步为通过发酵生产用于营养保健品和化妆品应用的商业白藜芦醇的菌株改良开辟了道路。
