促进桑黄次生代谢物累积的碳氮比及碳氮源优化研究
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摘要
为促进桑黄次生代谢物累积,以仅添加碳源和氮源的培养基培养桑黄菌丝体,采用比色法测定菌丝中黄酮、多酚、甾醇以及三萜含量。结果表明:碳氮比在60∶1～100∶1的试验范围内,黄酮、多酚和甾醇含量均在碳氮比为80∶1时达到最高,其单位含量分别为168.27,6.70,7.95mg·g~(-1),而三萜含量在碳氮比为100∶1时达最高,为4.17mg·g~(-1);在此基础上,优化了蔗糖、果糖、葡萄糖和乳糖为碳源和硝酸铵、碳酸铵和硝酸钠作为氮源的碳氮组合,黄酮、多酚和甾醇含量均在葡萄糖和碳酸铵组合下达到最高,其单位含量分别为251.45,4.68,10.48mg·g~(-1),而三萜含量在乳糖和碳酸铵组合下达到最高,为35.78mg·g~(-1)。由上述结果可知,适合桑黄菌丝次生代谢物积累的碳氮比为80∶1,最佳碳源和氮源分别为葡萄糖和碳酸铵。
In order to optimize the carbon and nitrogen sources and C/N ratios on secondary metabolites accumulation in Phellinus igniarius.The contents of flavonoids,polyphenols,sterols and triterpenoids in the mycelia of Phellinus igniarius were determined by colorimetry.The results showed that within the range of 60∶1 to100∶1 for C/N,the content of flavones,polyphenols and sterols all reached the highest when the ratio of C/N was 80∶1,which was 168.27,6.70 and 7.95 mg·g~(-1),respectively.While the triterpenoids content reached the highest when the ratio of C/N was was 100∶1,which was 4.17 mg·g~(-1);On this basis,we further analyzed the effect of different combinations with carbon sources(sucrose,fructose,glucose and lactose)and nitrogen sources(ammonium nitrate,ammonium carbonate and sodium nitrate)on the accumulation of flavonoids,polyphenols,sterols and triterpenoids.The results showed that the content of flavones,polyphenols and sterols all reached the highest under the combinations with glucose and ammonium carbonate,which was 251.45,4.68 and 10.48 mg·g~(-1),respectively.The content of triterpenoids reached the highest under the combinations with lactose and ammonium carbonatelucose.According to the above results,the suitable ratio of C/N for the accumulation of secondary metabolites was 80∶1,and the optimal carbon source and nitrogen source were glucose and ammonium carbonate,respectively.
In order to optimize the carbon and nitrogen sources and C/N ratios on secondary metabolites accumulation in Phellinus igniarius.The contents of flavonoids,polyphenols,sterols and triterpenoids in the mycelia of Phellinus igniarius were determined by colorimetry.The results showed that within the range of 60∶1 to100∶1 for C/N,the content of flavones,polyphenols and sterols all reached the highest when the ratio of C/N was 80∶1,which was 168.27,6.70 and 7.95 mg·g~(-1),respectively.While the triterpenoids content reached the highest when the ratio of C/N was was 100∶1,which was 4.17 mg·g~(-1);On this basis,we further analyzed the effect of different combinations with carbon sources(sucrose,fructose,glucose and lactose)and nitrogen sources(ammonium nitrate,ammonium carbonate and sodium nitrate)on the accumulation of flavonoids,polyphenols,sterols and triterpenoids.The results showed that the content of flavones,polyphenols and sterols all reached the highest under the combinations with glucose and ammonium carbonate,which was 251.45,4.68 and 10.48 mg·g~(-1),respectively.The content of triterpenoids reached the highest under the combinations with lactose and ammonium carbonatelucose.According to the above results,the suitable ratio of C/N for the accumulation of secondary metabolites was 80∶1,and the optimal carbon source and nitrogen source were glucose and ammonium carbonate,respectively.
引文
[1]夏国华,戈延茹,傅海珍,等.超声法提取桑黄总黄酮的工艺研究[J].江苏大学学报(医学版),2010,20(1):40-41.
[2]李文仙,俞丹,林玲,等.Folin-Ciocalteu比色法应用于蔬菜和水果总多酚含量测定的研究[J].营养学报,2011,33(3):302-307.
[3]傅海庆,陈绍军,陈汉清,等.珍稀药用菌桑黄的研究现状[J].福建农业学报,2005(20):117-120.
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[5]Wang Z Y,Wang C Y,Quan Y.Extraction of polysaccharides fromPhellinus nigricans mycelia and their antioxidant activities in vitro[J].Carbohydrate Polymers,2014,99(2):110-115.
[6]Zang Y,Xiong J,Zhai W Z,et al.Fomentarols A-D,sterols from the polypore macrofungus fomes fomentarius[J].Phytochemistry,2013,92:137-145.
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[8]曾晓希,周洪波,符波,等.灵芝多糖液体发酵条件的研究[J].现代生物医学进展,2007(6):830-832.
[9]林辰壹,马娟,杨婷婷,等.优化氮源种类及碳氮比对阿魏菇液体种生长的效应[J].新疆农业科学,2012,49(11):2042-2047.
[10]王振河,武忠伟,赵现方,等.营养物质对桑黄菌丝生物量及胞外多糖产量的影响[J].中国野生植物资源,2009,28(1):37-40.
[11]夏国华,戈延茹,傅海珍,等.超声法提取桑黄总黄酮的工艺研究[J].江苏大学学报(医学版),2010,20(1):40-41.
[12]李文仙,俞丹,林玲,等.Folin-Ciocalteu比色法应用于蔬菜和水果总多酚含量测定的研究[J].营养学报,2011,33(3):302-307.
[13]高虹,谷文英.三波长光度法测定姬松茸中麦角甾醇含量[J].分析化学,2007,35(4):586-588.
[14]李晓灿,詹亚光,王晓东,等.多胺对白桦悬浮细胞生长和三萜积累的影响[J].中草药,2013(4):463-467.
[15]刘淼,杨天灵,林辰壹.阿瓦提黄伞LL-01培养基碳氮比优化试验[J].食用菌,2014,36(4):29-30.
[16]张宇.不同碳氮比栽培料对秀珍菇菌丝及子实体生长的影响[J].北方园艺,2013(3):152-154.
[17]赵秀芳.白灵菇菌丝对不同碳氮源利用的研究[J].土壤肥料,2005(5):54-55.
[18]李翠翠,尉玉晓,郭立忠.桑黄液体发酵培养基优化的初步研究[J].中国食用菌,2009,28(2):46-48.
[19]孙荟林,李明,李守勉,等.茶树菇母种培养基最适碳氮比及碳源、氮源筛选[J].北方园艺,2016(14):148-151.
[20]孟丽,宋超群,吴宝杰,等.红平菇液体发酵培养基优化[J].食品研究与开发,2018,39(6):169-171.
[21]郭炜,于洪久,张楠,等.不同碳氮源对秀珍菇菌丝生长的影响[J].黑龙江农业科学,2018(8):82-84.
[22]Hur H.Cultural Characteristics and log-mediated cultivation of the medicinal mushroom,Phellinus linteus[J].Mycobiology,2008,36(2):81-87.
[2]李文仙,俞丹,林玲,等.Folin-Ciocalteu比色法应用于蔬菜和水果总多酚含量测定的研究[J].营养学报,2011,33(3):302-307.
[3]傅海庆,陈绍军,陈汉清,等.珍稀药用菌桑黄的研究现状[J].福建农业学报,2005(20):117-120.
[4]回晶,李辉,朱春玉,等.桑黄子实体与菌丝体营养成分的比较分析[J].特产研究,2009(2):59-61.
[5]Wang Z Y,Wang C Y,Quan Y.Extraction of polysaccharides fromPhellinus nigricans mycelia and their antioxidant activities in vitro[J].Carbohydrate Polymers,2014,99(2):110-115.
[6]Zang Y,Xiong J,Zhai W Z,et al.Fomentarols A-D,sterols from the polypore macrofungus fomes fomentarius[J].Phytochemistry,2013,92:137-145.
[7]黄丽洋,石卉,王晓婷,等.野生桑黄菌株的分离、鉴定及次生代谢物分析[J].中草药,2013,44(23):3394-3399.
[8]曾晓希,周洪波,符波,等.灵芝多糖液体发酵条件的研究[J].现代生物医学进展,2007(6):830-832.
[9]林辰壹,马娟,杨婷婷,等.优化氮源种类及碳氮比对阿魏菇液体种生长的效应[J].新疆农业科学,2012,49(11):2042-2047.
[10]王振河,武忠伟,赵现方,等.营养物质对桑黄菌丝生物量及胞外多糖产量的影响[J].中国野生植物资源,2009,28(1):37-40.
[11]夏国华,戈延茹,傅海珍,等.超声法提取桑黄总黄酮的工艺研究[J].江苏大学学报(医学版),2010,20(1):40-41.
[12]李文仙,俞丹,林玲,等.Folin-Ciocalteu比色法应用于蔬菜和水果总多酚含量测定的研究[J].营养学报,2011,33(3):302-307.
[13]高虹,谷文英.三波长光度法测定姬松茸中麦角甾醇含量[J].分析化学,2007,35(4):586-588.
[14]李晓灿,詹亚光,王晓东,等.多胺对白桦悬浮细胞生长和三萜积累的影响[J].中草药,2013(4):463-467.
[15]刘淼,杨天灵,林辰壹.阿瓦提黄伞LL-01培养基碳氮比优化试验[J].食用菌,2014,36(4):29-30.
[16]张宇.不同碳氮比栽培料对秀珍菇菌丝及子实体生长的影响[J].北方园艺,2013(3):152-154.
[17]赵秀芳.白灵菇菌丝对不同碳氮源利用的研究[J].土壤肥料,2005(5):54-55.
[18]李翠翠,尉玉晓,郭立忠.桑黄液体发酵培养基优化的初步研究[J].中国食用菌,2009,28(2):46-48.
[19]孙荟林,李明,李守勉,等.茶树菇母种培养基最适碳氮比及碳源、氮源筛选[J].北方园艺,2016(14):148-151.
[20]孟丽,宋超群,吴宝杰,等.红平菇液体发酵培养基优化[J].食品研究与开发,2018,39(6):169-171.
[21]郭炜,于洪久,张楠,等.不同碳氮源对秀珍菇菌丝生长的影响[J].黑龙江农业科学,2018(8):82-84.
[22]Hur H.Cultural Characteristics and log-mediated cultivation of the medicinal mushroom,Phellinus linteus[J].Mycobiology,2008,36(2):81-87.