清肺颗粒固态发酵培养基成分研究

doi:10.16431/j.cnki.1671-7236.2022.03.039

摘要/Abstract

摘要： 【目的】筛选最佳的清肺颗粒固态发酵培养基碳源、氮源、无机盐种类，优化清肺颗粒发酵培养基组成，提高组方中（R，S）-告依春的含量。【方法】建立发酵物中（R，S）-告依春含量的高效液相色谱法（HPLC）检测方法；单因素试验筛选清肺颗粒固态发酵培养基的碳源（葡萄糖、蔗糖、甘露醇、麦芽糖、玉米面、可溶性淀粉和清肺组方中药）、氮源（蛋白胨、黄豆粉、麸皮、酵母浸膏、硫酸铵、氯化铵和尿素）和无机盐（碳酸钙、磷酸二氢钾、硫酸镁、硫酸锰和氯化钠）成分，HPLC测定每克固态发酵培养物中（R，S）-告依春含量；4因素（碳源、氮源、无机盐和水）3水平响应面分析优化各培养基成分比例，用筛选出的最适培养基成分替代基础培养基中相应物质，接种解淀粉芽孢杆菌，于37 ℃、130 r/min振荡培养22 h，再以每克固态发酵培养物中（R，S）-告依春含量为评价指标，以响应面分析筛选培养基各组分含量。【结果】清肺颗粒最适固态发酵培养基碳源为组方中药粉，无机盐为CaCO₃，氮源为黄豆粉以及水；响应面分析优化的各培养基成分含量为：中药粉30%、黄豆粉10%、CaCO₃ 0.2%和水59.8%，此时固态发酵物中（R，S）-告依春含量达0.0184 mg/g，而未优化前清肺颗粒组方中（R，S）-告依春含量为0.0150 mg/g，二者差异极显著（P<0.01），说明解淀粉芽孢杆菌的发酵工艺更有利于清肺颗粒组方有效成分的释放。【结论】优化的固态发酵培养基易于解淀粉芽孢杆菌生长。

关键词: 清肺颗粒; 固态发酵; 培养基; (R,S)-告依春

Abstract: 【Objective】 The purpose of the experiment was to screen the best kinds of carbon source, nitrogen source and inorganic salt of Qingfei granule solid-state fermentation medium, optimize the composition of Qingfei fermentation medium and improve the content of epigoitrin in the formula.【Method】 A high performance liquid chromatography (HPLC) method was established for the determination of epigoitrin in fermented products.The carbon source (glucose, sucrose, mannitol, maltose, corn flour, soluble starch and herb), nitrogen source (peptone, soybean powder, bran, yeast extract, ammonium sulfate, ammonium chloride and urea) and inorganic salt(calcium carbonate, potassium dihydrogen phosphate, magnesium sulfate, manganese sulfate and sodium chloride) components of Qingfei granule solid-state fermentation medium were screened by single factor test.The content of epigoitrin in each gram of solid-state fermentation medium was determined by HPLC.Four factors (carbon source, nitrogen source, inorganic salt and water) and three levels response surface analysis was used to optimize the proportion of each medium component.The selected optimal medium component was used to replace the corresponding substances in the basic medium.The fermentation bacterium Bacillus amyloliquefacines was inoculated and cultured at 37 ℃ and 130 r/min for 22 h.Then the content of each component of the medium was analyzed by response surface analysis with the content of epigoitrin in each gram of solid fermentation culture as the evaluation index.【Result】 The optimum solid-state fermentation medium for Qingfei granules was traditional Chinese medicine powder, inorganic salt was calcium carbonate, and nitrogen source was soybean powder.The content of each medium optimized by response surface analysis was 30% traditional Chinese medicine powder, 10% of soybean powder, 0.2% of CaCO₃ and 59.8% of water.At this time, the content of epigoitrin in solid-state fermentation was 0.0184 mg/g.However, the content of epigoitrin in the prescription of Qingfei granules before optimization was 0.0150 mg/g, with extremely significant difference (P<0.01), it indicated that the fermentation process of Bacillus amyloliquefaciens was more conducive to the release of effective components in the prescription of Qingfei granules.【Conclusion】 The optimized solid-state fermentation substrate was easy for the growth of Bacillus amyloiquefaciens.

Key words: Qingfei granule; solid-state fermentation; culture medium; epigoitrin

中图分类号:

S853.7

张艳, 薛沾枚, 刘雪松, 苏景, 郝敬友, 唐伟, 刘彦凯, 孙洪杰, 史同瑞. 清肺颗粒固态发酵培养基成分研究[J]. 中国畜牧兽医, 2022, 49(3): 1162-1171.

ZHANG Yan, XUE Zhanmei, LIU Xuesong, SU Jing, HAO Jingyou, TANG Wei, LIU Yankai, SUN Hongjie, SHI Tongrui. Study on Components of Solid-state Fermentation Medium of Qingfei Granules[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 1162-1171.

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