灵芝菌深层培养及灵芝多糖分离纯化的研究

【作者】 罗立新；

【导师】 姚汝华；

【作者基本信息】 华南理工大学 ， 发酵工程， 1996， 博士

【摘要】 灵芝是我国传统的名贵中药，一直被人们用来治疗肝病、肾炎、胃溃疡、高血压、关节炎、神经衰弱、支气管炎、哮喘等疾病，市场需求量与日俱增。利用深层培养技术，大规模地生产灵芝菌丝体，并从中获取有效成分具有周期短、操作方便、成本低、产量大的特点，已成为当前名贵真菌开发的热点之一。本论文对灵芝菌种选育，深层培养条件及发酵动力学，灵芝多糖的分离、纯化方法等方面进行了较为全面和深入的研究和探讨，并对灵芝多糖的化学结构进行了测定，其结果表明： 1．采用紫外线对菌株进行了诱变处理，选育到一株突变菌株UV—60S，其菌体产量和胞外粗多糖含量分别比原菌株提高了21.3％和30.6％。 2．研究了碳、氮源，无机盐，生长促进剂等因素对菌体生长的影响。对发酵条件的影响也进行了分析，确定了最佳发酵培养基为： 葡萄糖5％、玉米浆0.3％、豆饼粉1.0％、糖蜜0.6％、KH₂PO₄·3H₂O 0.15％、MgSO₄·7H₂O 0.075％、三十烷醇1ppm。 最佳摇瓶发酵条件为：种龄72-96hr、接种量5％、装液量50ml、摇床转速150r／min、温度28℃、初始PH 5.5-6.0。 最佳发酵工艺条件为：温度28℃、搅拌转速150—200r／min、装液量3升、通风量3—5升／分钟、初始PH 5.5。 在最佳发酵工艺条件下，菌丝体千重可达15.1g／L，胞外粗多糖含量达760mg／L，分别比工艺优化前提高了15.3％和18.8％。 3．根据灵芝菌丝体发酵过程的代谢特征，经研究建立了基于逻辑方程和Luedeking—Piret方程的动力学模型： dx／dt=μ_mX（1-X／X_m），（t≥48hr） dp／dt=m₁X+m₂（dx／dt） -ds／dt=b₁X+b₂（dx／dt）+b₃（dp／dt）模型的预测值和实验值之间的平均相对偏差小于10％，较好地描述了菌体生长的变化规律。更多还原

【Abstract】 Ganoderma lucidum (GL) has long been a precious Chinese medicine to cure various human diseases, such as hepato-pathy, nephritis, gastric ulcer, hypertension, arthritis, neurasthenia, bronchitis, asthma. In recent years, there is a great demand for Ganoderma lucidum. The submerged culture has the characteristics of short production cycle, high yield, low cost and easy to operate, so it’s becoming much more popular to exploit fungi resources. This paper has made comparatively through exploration on the breeding of strain, fermentation conditions, fermentation kinetics, separation and purification of Ganoderma lucidum Polysaccharide (GLP), the structure of this polysaccharide was also determined. Those results showed:1. The strain AS.5.65 was mutated by UV-rays, UV-60S, as one of mutant strains, which mycelial yield and extracellular polysaccharide content increased by 21.3% and 30.6%.2. The effects of culture conditions (i.e. carbon sources, nitrogen sources, metal ions, growth factor, temperature, dissolved oxygen and PH) on the mycelial growth of GL were investigated, the optimum media and conditions were also selected:The optimum medium: glucose 5%, corn-steep liquor 0.3%, soybean cake powder 1.0%, waste molasses 0.6%, KH2PO4 · 3H2O 0.15%, MgSO4 · 7H2O 0.075%, triacontanol 1ppm.The optimum flask culture conditions: inoculum age 72-96hr, inoculum size 5%, volume of medium 50ml, shake frequency 150r/min, temperature 28℃, PH 5.5 - 6.0.The culture conditions: temperature 28℃, agitation speed 150-200r/min, volume of medium 3L, ventilation 3-5L/min, PH 5.5.Under the optimum culture conditions, the dry weight of mycelia reached 15.1g/L, was raised by 15.3% and the content of extracellular polysaccharide 760mg/L, 18.8% higher than the control.更多还原