Ghatnur Shashidhar M, Parvatam Giridhar, Balaraman Manohar
Academy of Scientific and Innovative Research, CSIR-CFTRI, Mysore, Karnataka, India; Department of Food Engineering, CSIR-Central Food Technological Research Institute, Mysore, Karnataka, India.
Department of Plant Cell Biotechnology, CSIR-Central Food Technological Research Institute, Mysore, Karnataka, India.
Pharmacogn Mag. 2015 Oct;11(Suppl 3):S448-56. doi: 10.4103/0973-1296.168946.
Cordyceps sinensis (CS) is a traditional Chinese medicine contains potent active metabolites such as nucleosides and polysaccharides. The submerged cultivation technique is studied for the large scale production of CS for biomass and metabolites production.
To optimize culture conditions for large-scale production of CS1197 biomass and metabolites production.
The CS1197 strain of CS was isolated from dead larvae of natural CS and the authenticity was assured by the presence of two major markers adenosine and cordycepin by high performance liquid chromatography and mass spectrometry. A three-level Box-Behnken design was employed to optimize process parameters culturing temperature, pH, and inoculum volume for the biomass yield, adenosine and cordycepin. The experimental results were regressed to a second-order polynomial equation by a multiple regression analysis for the prediction of biomass yield, adenosine and cordycepin production. Multiple responses were optimized based on desirability function method.
The desirability function suggested the process conditions temperature 28°C, pH 7 and inoculum volume 10% for optimal production of nutraceuticals in the biomass. The water extracts from dried CS1197 mycelia showed good inhibition for 2 diphenyl-1-picrylhydrazyl and 2,2-azinobis-(3-ethyl-benzo-thiazoline-6-sulfonic acid-free radicals.
The result suggests that response surface methodology-desirability function coupled approach can successfully optimize the culture conditions for CS1197.
Authentication of CS1197 strain by the presence of adenosine and cordycepin and culturing period was determined to be for 14 daysContent of nucleosides in natural CS was found higher than in cultured CS1197 myceliumBox-Behnken design to optimize critical cultural conditions: temperature, pH and inoculum volumeWater extract showed better antioxidant activity proving credible source of natural antioxidants.
冬虫夏草是一种含有核苷和多糖等强效活性代谢物的传统中药。人们对深层发酵培养技术进行了研究,以大规模生产冬虫夏草用于生物量和代谢物的生产。
优化培养条件,以大规模生产冬虫夏草CS1197的生物量和代谢物。
从天然冬虫夏草的死幼虫中分离出冬虫夏草CS1197菌株,并通过高效液相色谱和质谱法检测到两种主要标志物腺苷和虫草素,从而确保其真实性。采用三水平Box-Behnken设计优化培养温度、pH值和接种量等工艺参数,以提高生物量产量、腺苷和虫草素含量。通过多元回归分析将实验结果回归为二阶多项式方程,以预测生物量产量、腺苷和虫草素的产量。基于期望函数法对多个响应进行优化。
期望函数表明,温度28℃、pH值7和接种量10%的工艺条件有利于在生物量中最佳生产营养保健品。干燥的CS1197菌丝体的水提取物对2,2-二苯基-1-苦基肼基和2,2'-联氮双(3-乙基苯并噻唑啉-6-磺酸)自由基显示出良好的抑制作用。
结果表明,响应面法-期望函数耦合方法可以成功优化CS1197的培养条件。
通过腺苷和虫草素的存在对CS1197菌株进行鉴定,培养周期确定为14天。发现天然冬虫夏草中的核苷含量高于培养的CS1197菌丝体。采用Box-Behnken设计优化关键培养条件:温度、pH值和接种量。水提取物显示出更好的抗氧化活性,证明是天然抗氧化剂的可靠来源。