甘草的综合开发的研究

【作者】 陈永强；

【导师】 孙启玲；

【作者基本信息】 四川大学 ， 微生物学， 2007， 硕士

【摘要】 甘草为豆科植物甘草Glycyrrhiza uralensis Fisch．的根及根状茎，具有清热解毒、止咳祛痰、补脾和胃、调和诸药的功效。其主要有效成分为三萜皂苷甘草酸，含量在3.7—8.3％。其皂苷元甘草次酸也是甘草中重要有效成分之一，含量较低，只有0.1％左右。本论文的主要研究内容和结果：Ⅰ采用复杂系统定向调控技术和常规单因素实验研究甘草发酵酶解工艺。最终确立了微生物发酵甘草生产甘草次酸的发酵酶解的最优工艺为：发酵时间3d，酶解时间10h，酶解pH为5.0，酶解温度为40℃；Ⅱ对甘草煮提液发酵动力学进行研究，通过检测发酵过程中菌体生长与营养物质消耗、酶活变化与转化率之间的关系，阐明微生物发酵转化甘草的机理，为甘草发酵条件的优化提供指导；Ⅲ采用单因素和正交实验对甘草酸的发酵的种子和转化工艺进行研究。得到最佳的条件为：种子培养：玉米糖化液60％，甘草酸粗品0.1％，麸皮汁0.5％，酵母膏1.6％，无机盐浓缩液少许，接种量为0.2％（孢子悬液浓度为1.0×10⁸），32℃培养24h；摇瓶培养：转速170rpm，前24小时采用32℃，以后温度35℃，接种量30％，转化浓度1.5％，装液量30％，转化时间72小时；发酵罐培养：转速250rpm，前24小时采用32℃，以后温度35℃，接种量30％，转化浓度1.5％，装液量60％，通气量0.1m³／h，转化时间：72小时。Ⅳ甘草的综合开发：采用正交实验研究甘草废渣中的黄酮的氢氧化钙提取工艺，确定提取工艺为：Ca（OH）₂：甘草渣=0.045：1；固液比即加入水量：甘草渣=30 ml：1g；浸提温度为100℃；浸提时间3h。余下残渣进行亮菌饲料发酵。甘草多糖的提取工艺：提取甘草次酸后的发酵液（10％）500ml，浓缩至原体积的1／5，过滤，除去水不溶性物，取滤液，用1mol／l的盐酸调pH=2，沉淀12小时，离心除去果胶、部分黄酮、少量的甘草皂苷类成分。滤液用1mol／l的NaOH调至中性后继续浓缩至原体积的1／3。向浓缩液中加入3倍量95％的乙醇，于冰箱中放置醇析过夜。离心，收集沉淀，低温60℃烘干，即得甘草多糖。更多还原

【Abstract】 The roots of Glycyrrhiza uralensis Fisch, commonly known as liquorice, havetherapeutic properties. The main component is triterpenoid saponin thetriterpeneglycyrrhizinic acid（GL） which is between 3% and 8%. Another bioactive component isglycyrrhetinic acid （GA） which is mainly used in pharmaceutical and cosmetic fields asanti-inflammatory, anti-ulcer activities and anti reddeningagents. Furthermore, theanti-hepatotoxic and the hypolipidemic activity of GA have been demonstrated. But thecontent of GA in liquorice is only about 0.1%.The main contents and results include:ⅠWe have easily isolated HC-12 from soil samples of wild Glycyrrhiza inXinjiang province. Through the analysis of TLC and HPLC Results, HC—12 isAspergillus oryzae with high ability on conversing of GL.ⅡThe fermentation kinetics of liquorice, including the growth characteristics of A.oryzae HC-12 in batch culture, together with the change of the amount of GL and GA,the activity ofβ-glucuronidase during the fermentation, were monitoredsimultaneously.ⅢOptimization for the ferment conditions include:1. Culture medium of the germ: lixivium of corn 60%, GL 0.1%, lixivium of bran 0.5%,solution of salt a little;2. The optimized medium conditions in shake flask culture by using an orthogonallayout were the speed of rotation is 170rpm, the concentration of GL is 1.5%, cultivatetemperature of 32℃at begin 24 hours and then 35℃, inoculum size of 30% andculturing time for 72 hr.3. The optimized medium conditions in fermenter is the concentration of GL is1.5%, cultivate temperature of 32℃at begin 24 hours and then 35℃, inoculum size of30% and culturing time for 72 hr. ⅣThe overall study of liquorice. The optimized extract conditions of liquiritin isin shake flask culture by using an orthogonal layout were the Ca（OH）₂: liquiritinresidue=0.045: 1, liquiritin residue: water=1:30, extract temperature 100℃, theextract time 2 h.更多还原