微生物多级发酵对中药方剂功能活性及有效组分的影响

Effect of Microbial Multistage Fermentation on Functional Activity and Effective Components of Traditional Chinese Medicine Prescription

摘要:

选用多菌株对一中药方剂（简称JGZF）进行液体多级发酵，考察其在3种处理方式（直接熬制、自然发酵、人工发酵）下微量元素、体外抗氧化活性、抑菌活性以及其他有效组分等结果的变化。采用原子吸收光谱法检测微量元素；紫外-可见分光光度计检测体外抗氧化活性，包括过氧化物歧化酶SOD活性、DPPH自由基的清除能力、羟基自由基的清除能力和总多酚含量；牛津杯抑菌圈法测定抑菌活性；UPLC-Q-TOF-MS/MS非靶标代谢组学检测JGZF的药效成分。结果表明：跟直接熬制相比，中药方剂JGZF经自然发酵和人工发酵处理后，锌、铜、铁和钙的含量得到了显著提高；人工发酵能显著提高JGZF的体外抗氧化活性，其中SOD活性提高了1.5倍，DPPH自由基的清除能力提高了2.2倍，羟基自由基的清除能力提高了4.2倍，总多酚含量提高了2.2倍；发酵后的JGZF对沙门氏菌和大肠杆菌的抑菌活性更强，但对志贺氏菌和金黄色葡萄球菌的抑菌活性不变或略有降低；JGZF经人工发酵后，药液组分含量有增有减，其中人体必需氨基酸L-苯丙氨酸、抗炎物质黄芩素等含量显著增加；五味子苷B、7-羟基香豆素、农药残留等含量有显著降低。

Abstract:

The liquid multistage fermentation of a traditional Chinese medicine prescription (JGZF) was carried out with multiple strains. The changes of trace elements, antioxidant activity in vitro, antibacterial activity and other effective components of JGZF were investigated under three treatment methods (direct boiling, natural fermentation and artificial fermentation). The trace elements were detected by atomic absorption spectrometry, and the antioxidant activities in vitro were detected by ultraviolet visible spectrophotometer, including the superoxide dismutase(SOD) activity, DPPH radical scavenging capacity, hydroxyl radical scavenging capacity and the total polyphenol content. The antibacterial activity was determined by Oxford cup method and evaluated based on the diameter size of bacteriostatic ring. The pharmacodynamic components of JGZF were detected by UPLC-Q-TOF-MS/MS non-target metabonomics detection. The results showed that the contents of trace elements including Zn, Cu, Fe and Ca in JGZF after the natural fermentation and artificial fermentation were significantly increased compared with those of direct boiling. The antioxidant activity in vitro of JGZF was significantly improved by artificial fermentation, in which SOD activity was increased by 1.5 times, DPPH radical scavenging capacity was increased by 2.2 times, hydroxyl radical scavenging ability was increased by 4.2 times, and the content of total polyphenols increased by 2.2 times. The antibacterial activity of JGZF against Salmonella and Escherichia coli was stronger, however, the antibacterial activity against Shigella and Staphylococcus aureus unchanged or slightly decreased. After the artificial fermentation of JGZF, the content of liquid components increased or decreased, including the essential amino acid L-phenylalanine, anti-inflammatory substances such as baicalein increased by hundreds of millions of times, while the contents of huoxuetong schisandrin B, 7-hydroxycoumarin and pesticide residue were significantly decreased.