发酵制备冬虫夏草饲料添加剂的工艺研究
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摘要
冬虫夏草是一种珍贵的药物真菌,具有许多药理功能,但天然的冬虫夏草产量十分稀少,因此,人们采用培养冬虫夏草菌丝体来替代天然冬虫夏草。目前,人工虫草菌丝体的生产多以液体深层发酵为主。本论文以农产副产物米糠和豆粕作为固体发酵的培养基,以虫草活性成分β-葡聚糖的产率为发酵指标,对冬虫夏草固体发酵生产饲料添加剂技术进行了研究。
液态发酵中实验以虫草菌丝体及胞外多糖的产量为衡量标准,对几种碳源及氮源进行选择,确定了以蔗糖作为碳源,酵母膏作为氮源。通过单因素实验和正交试验,对液体培养基组分浓度进行优化,结果表明:蔗糖20g/L,酵母膏2g/L,磷酸二氢钾1g/L,硫酸镁0.6g/L为较适的培养基条件。通过对发酵条件影响因素:接种量、温度、培养时间和通气量等研究,确定了在蔗糖、酵母膏液体培养基下发酵生产多糖和菌丝体的最佳条件为:接种量7%、温度26℃、培养时间4d和通气流量1L/min。以最优条件培养得到菌丝体及多糖的产量为12.35g/L和25.10g/L。
以豆粕和米糠为固态培养基,固体发酵生产冬虫夏草多糖。通过对发酵生产冬虫夏草多糖条件的影响因素:包括培养基组成配比、接种量、温度、培养基相对湿度、空气相对湿度和培养时间的单因素及正交实验优化的研究,初步确定了在豆粕:米糠为1:2(w/w)固体培养基发酵生产冬虫夏草多糖的最佳条件为:接种量25%、温度26℃、培养基含水量为60%、空气相对湿度为60%和培养时间为8d。以最优条件培养得到的1g菌粉中葡聚糖产量为17.03mg。此外,经傅立叶红外光谱分析、薄层色谱及核磁共振分析,可以确定本研究生产的冬虫夏草多糖为β-D葡聚糖。
Cordyceps sinensis is a famous fungus which has many pharmacological features. Considering the scarce yield of natural Cordyceps sinensis,it's very important to proceed the studies on producing Cordyceps sinensis mycelium.At present,Artificial Cordyceps mycelium production in submerged fermentation and more dominated.In this paper,the production technology of the feed additive Cordyceps sinensis by solid state fermentation were studied,which Cordyceps active ingredients ofβ-glucan with the optimized was as bacteria,rice bran and soybean meal as the solid state fermentation medium.
The Cordyceps sinensis mycelia and polysaccharides were prepared by liquid fermentation.I choose several carbon and nitrogen sources and then identified sucrose as a carbon source,yeast extract as nitrogen source.Through single factor experiments and orthogonal test,the optimum liquid medium components were as follows:sucrose 20g/L, yeast extract 2g/L,potassium dihydrogen phosphate 1 g/L,magnesium sulfate 0.6g/L. The effects of fermentation conditions including the quantity of inoculation,cultural temperature,fermentation time and volume of flux air on the yield of the mycelia and polysaccharides were researched.The optimum conditions were as follows:7%of inoculum concentration,26℃of temperature,4 days of fermentation time and 1L/min of air flow speed.Mycelium and polysaccharide production are 12.35g/L and 25.10g/L in optimal conditions.
The Cordyceps sinensis polysaccharide was prepared by solid state fermentations, which the soybean meal and rice bran were as raw materials.The effects of solid fermentation conditions including the quantity of inoculation,cultural temperature, substrate water content,relative humidity and fermentation time were researched.The optimum conditions were as follows:25%of inoculum concentration,26℃of temperature, 60%of substrate water content,60%of relative humidity and 8 days of fermentation time. In addition,the structure of the produced Cordyceps sinensis polysaccharides was analyzed by the Fourier Transform Infrared Spectroscopy,TLC,nuclear magnetic resonance and the polysaccharide was seemed toβ-glucan.
液态发酵中实验以虫草菌丝体及胞外多糖的产量为衡量标准,对几种碳源及氮源进行选择,确定了以蔗糖作为碳源,酵母膏作为氮源。通过单因素实验和正交试验,对液体培养基组分浓度进行优化,结果表明:蔗糖20g/L,酵母膏2g/L,磷酸二氢钾1g/L,硫酸镁0.6g/L为较适的培养基条件。通过对发酵条件影响因素:接种量、温度、培养时间和通气量等研究,确定了在蔗糖、酵母膏液体培养基下发酵生产多糖和菌丝体的最佳条件为:接种量7%、温度26℃、培养时间4d和通气流量1L/min。以最优条件培养得到菌丝体及多糖的产量为12.35g/L和25.10g/L。
以豆粕和米糠为固态培养基,固体发酵生产冬虫夏草多糖。通过对发酵生产冬虫夏草多糖条件的影响因素:包括培养基组成配比、接种量、温度、培养基相对湿度、空气相对湿度和培养时间的单因素及正交实验优化的研究,初步确定了在豆粕:米糠为1:2(w/w)固体培养基发酵生产冬虫夏草多糖的最佳条件为:接种量25%、温度26℃、培养基含水量为60%、空气相对湿度为60%和培养时间为8d。以最优条件培养得到的1g菌粉中葡聚糖产量为17.03mg。此外,经傅立叶红外光谱分析、薄层色谱及核磁共振分析,可以确定本研究生产的冬虫夏草多糖为β-D葡聚糖。
Cordyceps sinensis is a famous fungus which has many pharmacological features. Considering the scarce yield of natural Cordyceps sinensis,it's very important to proceed the studies on producing Cordyceps sinensis mycelium.At present,Artificial Cordyceps mycelium production in submerged fermentation and more dominated.In this paper,the production technology of the feed additive Cordyceps sinensis by solid state fermentation were studied,which Cordyceps active ingredients ofβ-glucan with the optimized was as bacteria,rice bran and soybean meal as the solid state fermentation medium.
The Cordyceps sinensis mycelia and polysaccharides were prepared by liquid fermentation.I choose several carbon and nitrogen sources and then identified sucrose as a carbon source,yeast extract as nitrogen source.Through single factor experiments and orthogonal test,the optimum liquid medium components were as follows:sucrose 20g/L, yeast extract 2g/L,potassium dihydrogen phosphate 1 g/L,magnesium sulfate 0.6g/L. The effects of fermentation conditions including the quantity of inoculation,cultural temperature,fermentation time and volume of flux air on the yield of the mycelia and polysaccharides were researched.The optimum conditions were as follows:7%of inoculum concentration,26℃of temperature,4 days of fermentation time and 1L/min of air flow speed.Mycelium and polysaccharide production are 12.35g/L and 25.10g/L in optimal conditions.
The Cordyceps sinensis polysaccharide was prepared by solid state fermentations, which the soybean meal and rice bran were as raw materials.The effects of solid fermentation conditions including the quantity of inoculation,cultural temperature, substrate water content,relative humidity and fermentation time were researched.The optimum conditions were as follows:25%of inoculum concentration,26℃of temperature, 60%of substrate water content,60%of relative humidity and 8 days of fermentation time. In addition,the structure of the produced Cordyceps sinensis polysaccharides was analyzed by the Fourier Transform Infrared Spectroscopy,TLC,nuclear magnetic resonance and the polysaccharide was seemed toβ-glucan.
引文
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