蛹虫草菌丝体胞内多糖提取方法的研究
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摘要
以提取温度、料液比、提取时间和提取次数为因素,采用正交试验研究热水浸提的最佳提取条件,并以此为基础研究热水浸提、冷热交替浸提、盐酸溶液浸提和氢氧化钠溶液浸提对蛹虫草菌丝体胞内多糖提取率的影响.结果表明:热水浸提的最佳提取工艺为:m(料)∶V(液)=1∶40,提取温度75℃,提取时间2.5 h,提取2次,多糖的提取率为14.74%,其中料液比为影响多糖提取率的主要因素,其他因素依次是提取温度、提取时间和提取次数.按照此工艺,冷热交替浸提(热水浸提2.5 h+冷水浸提48 h)多糖的提取率最高,为15.92%. 55 g/L的氢氧化钠溶液浸提多糖的提取率为6.31%; 25 g/L的盐酸溶液浸提多糖的提取率为9.93%.因此,冷热交替浸提蛹虫草菌丝体胞内多糖的提取率最高,其次是热水浸提、盐酸溶液浸提和氢氧化钠溶液浸提.
The optimum condition of hot water extraction was investigated using an orthogonal array design L9(34)with extraction temperature,solid-solvent ratio,extraction time and number of extraction cycles as independent variables. The effects of hot water extraction,thermal cycling extraction,hydrochloric acid extraction and sodium hydroxide alkali extraction on the extraction rate of intracellular polysaccharides from Cordyceps militaris were studied to optimize the extraction parameters. The results showed that the optimum extraction conditions of hot water extraction of intracellular polysaccharides were as follows: solid-solvent ratio of 1(g) ∶ 40(m L),75 ℃,2 extraction cycles,and 2.5 h each cycle. The sequence of factors that worked on the extraction rate of polysaccharides is,in order of significance,solid-solvent ratio,extracting temperature,extracting time and extracting times. The optimum extracting condition of thermal cycling extraction method was hot water extraction for 2.5 h followed by cold water extraction for 48 h. The corresponding extraction rate of intracellular polysaccharides was 15.92%. Sodium hydroxide extraction(55 g/L) and hydrochloric acid extraction(25 g/L) exhibited the highest extraction rate of 6.31% and9.93% respectively. In conclusion,the optimum extraction method is thermal cycling extraction,followed by hot water extraction,hydrochloric acid extraction and sodium hydroxide extraction in turn.
The optimum condition of hot water extraction was investigated using an orthogonal array design L9(34)with extraction temperature,solid-solvent ratio,extraction time and number of extraction cycles as independent variables. The effects of hot water extraction,thermal cycling extraction,hydrochloric acid extraction and sodium hydroxide alkali extraction on the extraction rate of intracellular polysaccharides from Cordyceps militaris were studied to optimize the extraction parameters. The results showed that the optimum extraction conditions of hot water extraction of intracellular polysaccharides were as follows: solid-solvent ratio of 1(g) ∶ 40(m L),75 ℃,2 extraction cycles,and 2.5 h each cycle. The sequence of factors that worked on the extraction rate of polysaccharides is,in order of significance,solid-solvent ratio,extracting temperature,extracting time and extracting times. The optimum extracting condition of thermal cycling extraction method was hot water extraction for 2.5 h followed by cold water extraction for 48 h. The corresponding extraction rate of intracellular polysaccharides was 15.92%. Sodium hydroxide extraction(55 g/L) and hydrochloric acid extraction(25 g/L) exhibited the highest extraction rate of 6.31% and9.93% respectively. In conclusion,the optimum extraction method is thermal cycling extraction,followed by hot water extraction,hydrochloric acid extraction and sodium hydroxide extraction in turn.
引文
[1] SHRESTHA B,ZHANG W,ZHANG Y,et al. The medicinal fungus Cordyceps militaris:research and development[J]. Mycological Progress,2012,11(3):599-614.
[2]屈燕,区智.蛹虫草的研究进展[J].现代农业科技,2012(12):74-79.
[3] HUANG Z F,ZHANG M L,ZHANG S,et al. Structural characterization of polysaccharides from Cordyceps militaris and their hypolipidemic effects in high fat diet fed mice[J]. Rsc Advances,2018,8:41012-41022.
[4] LIU J Y,FENG C P,LI X,et al. Immunomodulatory andantioxidative activity of Cordyceps militaris polysaccharides in mice[J]. International Journal of Biological Macromolecules,2016,86:594-598.
[5] LIU F,ZHU Z Y,SUN X,et al. The preparation of three selenium-containing Cordyceps militaris polysaccharides:characterization and anti-tumor activities[J]. International Journal of Biological Macromolecules,2017,99:196-204.
[6] SONG J,WANG Y,TENG M,et al. Cordyceps militaris induces tumor cell death via the caspase-dependent mitochondrial pathway in Hep G2 and MCF-7 cells[J]. Molecular Medicine Reports,2016,13(6):5132-5140.
[7]陈晓鹏,汪振炯,华春,等.超声波-酶法提取蛹虫草基质多糖的工艺研究[J].食品科技,2014,39(3):142-147.CHEN X P,WANG Z J,HUA C,et al. Optimization of the extraction of Cordyceps militaris polysaccharides by combined ultrasonic wave treatment and enzyme hydrolysis[J]. Food Science and Technology,2014,39(3):142-147.
[8]张慧,罗帷.蛹虫草中虫草素的研究与开发进展[J].现代农业科技,2016(9):292-292.
[9]项玥,唐芳荣,昌艳,等.蛹虫草菌丝体胞内多糖热水浸提工艺探究[J].食品研究与开发,2008,29(8):12-15.XIANG Y,TANG F R,CHANG Y,et al. Optimization of the water extraction technique of Cordyceps militaris mycelium polysaccharides[J]. Food Research and Development,2008,29(8):12-15.
[10]王再花,章金辉,李杰,等. 3种药用石斛多糖的抗氧化活性比较研究[J].华南师范大学学报(自然科学版),2015,47(5):65-70.WANG Z H,ZHANG J H,LI J,et al. Comparison of antioxidant activity of the polysaccharides from three dendrobium species[J]. Journal of South China Normal University(Natural Science Edition),2015,47(5):65-70.
[11] XU N,REN Z,ZHANG J,et al. Antioxidant and anti-hyperlipidemic effects of mycelia zinc polysaccharides by Pleurotus eryngii var. tuoliensis[J]. International Journal of Biological Macromolecules,2017,95:204-214.
[12] LIU G Q,ZHANG H Y,WANG X L,et al. Extraction of mycelial polysaccharides from submerged-cultured Cordyceps militaris(a caterpillar fungus)[C]∥International Conference on Biomedical Engineering and Informatics. IEEE,2010:1946-1950.
[13]吴雅静,张名位,孙远明,等.荔枝多糖的超声波辅助提取工艺优化研究[J].华南师范大学学报(自然科学版),2007(2):119-124.WU Y J,ZHANG M W,SUN Y M,et al. Investigation the ultrasonic wave-assisted extraction technology for Litchi polysaccharides[J]. Journal of South China Normal University(Natural Science Edition),2007(2):119-124.
[14]谢红旗,周春山,杜邵龙,等.酶法提取、超滤分离香菇多糖新工艺研究[J].食品科学,2007,28(4):217-219.XIE H Q,ZHOU C S,DU S L,et al. Study on enzymatic extraction and ultrafiltration Separation of Lentinan[J].Food Science,2007,28(4):217-219.
[15]陈方圆,焦子伟,努尔买买提,等.蛹虫草活性物质提取技术研究进展[J].江苏农业科学,2017,45(6):7-13.
[16] DUBOIS M,GILLES K A,HAMILTON J K,et al. Colorimetric method for determination of sugars and relatedsubstances[J]. Analytical Chemistry,1956,28(3):350-356.
[17] ROBERTSON J A,RYDEN P,BOTHAM R L,et al.Structural properties of diet-derived polysaccharides and their influence on butyrate production during fermentation[J]. LWT-Food Science and Technology,2001,34(8):567-573.
[18] LI S Q,SHAH N P. Characterization,antioxidative and bifidogenic effects of polysaccharides from Pleurotus eryngii after heat treatments[J]. Food Chemistry,2016,197:240-249.
[19]苏红,李青连,魏磊,等.真菌多糖水提及化学辅助提取方法研究进展[J].微生物学通报,2010,37(3):426-432.SU H,LI Q L,WEI L,et al. Research progress in water extraction method and chemical assisted extraction method of fungal polysaccharides[J]. Microbiology China,2010,37(3):426-432.
[20] PEASURA N,LANHAKUNJIT N,KERDCHOECHUEN O,et al. Characteristics and antioxidant of Ulva intestinalis sulphated polysaccharides extracted with different solvents[J]. International Journal of Biological Macromolecules,2015,81(4):912-919.
[21]刘婷,周光明.多糖的提取和分析方法[J].化工时刊,2008,22(3):66-70.LIU T,ZHOU G M. Extraction and analytical method of polysaccharides[J]. Chemical Industry Times,2008,22(3):66-70.
[2]屈燕,区智.蛹虫草的研究进展[J].现代农业科技,2012(12):74-79.
[3] HUANG Z F,ZHANG M L,ZHANG S,et al. Structural characterization of polysaccharides from Cordyceps militaris and their hypolipidemic effects in high fat diet fed mice[J]. Rsc Advances,2018,8:41012-41022.
[4] LIU J Y,FENG C P,LI X,et al. Immunomodulatory andantioxidative activity of Cordyceps militaris polysaccharides in mice[J]. International Journal of Biological Macromolecules,2016,86:594-598.
[5] LIU F,ZHU Z Y,SUN X,et al. The preparation of three selenium-containing Cordyceps militaris polysaccharides:characterization and anti-tumor activities[J]. International Journal of Biological Macromolecules,2017,99:196-204.
[6] SONG J,WANG Y,TENG M,et al. Cordyceps militaris induces tumor cell death via the caspase-dependent mitochondrial pathway in Hep G2 and MCF-7 cells[J]. Molecular Medicine Reports,2016,13(6):5132-5140.
[7]陈晓鹏,汪振炯,华春,等.超声波-酶法提取蛹虫草基质多糖的工艺研究[J].食品科技,2014,39(3):142-147.CHEN X P,WANG Z J,HUA C,et al. Optimization of the extraction of Cordyceps militaris polysaccharides by combined ultrasonic wave treatment and enzyme hydrolysis[J]. Food Science and Technology,2014,39(3):142-147.
[8]张慧,罗帷.蛹虫草中虫草素的研究与开发进展[J].现代农业科技,2016(9):292-292.
[9]项玥,唐芳荣,昌艳,等.蛹虫草菌丝体胞内多糖热水浸提工艺探究[J].食品研究与开发,2008,29(8):12-15.XIANG Y,TANG F R,CHANG Y,et al. Optimization of the water extraction technique of Cordyceps militaris mycelium polysaccharides[J]. Food Research and Development,2008,29(8):12-15.
[10]王再花,章金辉,李杰,等. 3种药用石斛多糖的抗氧化活性比较研究[J].华南师范大学学报(自然科学版),2015,47(5):65-70.WANG Z H,ZHANG J H,LI J,et al. Comparison of antioxidant activity of the polysaccharides from three dendrobium species[J]. Journal of South China Normal University(Natural Science Edition),2015,47(5):65-70.
[11] XU N,REN Z,ZHANG J,et al. Antioxidant and anti-hyperlipidemic effects of mycelia zinc polysaccharides by Pleurotus eryngii var. tuoliensis[J]. International Journal of Biological Macromolecules,2017,95:204-214.
[12] LIU G Q,ZHANG H Y,WANG X L,et al. Extraction of mycelial polysaccharides from submerged-cultured Cordyceps militaris(a caterpillar fungus)[C]∥International Conference on Biomedical Engineering and Informatics. IEEE,2010:1946-1950.
[13]吴雅静,张名位,孙远明,等.荔枝多糖的超声波辅助提取工艺优化研究[J].华南师范大学学报(自然科学版),2007(2):119-124.WU Y J,ZHANG M W,SUN Y M,et al. Investigation the ultrasonic wave-assisted extraction technology for Litchi polysaccharides[J]. Journal of South China Normal University(Natural Science Edition),2007(2):119-124.
[14]谢红旗,周春山,杜邵龙,等.酶法提取、超滤分离香菇多糖新工艺研究[J].食品科学,2007,28(4):217-219.XIE H Q,ZHOU C S,DU S L,et al. Study on enzymatic extraction and ultrafiltration Separation of Lentinan[J].Food Science,2007,28(4):217-219.
[15]陈方圆,焦子伟,努尔买买提,等.蛹虫草活性物质提取技术研究进展[J].江苏农业科学,2017,45(6):7-13.
[16] DUBOIS M,GILLES K A,HAMILTON J K,et al. Colorimetric method for determination of sugars and relatedsubstances[J]. Analytical Chemistry,1956,28(3):350-356.
[17] ROBERTSON J A,RYDEN P,BOTHAM R L,et al.Structural properties of diet-derived polysaccharides and their influence on butyrate production during fermentation[J]. LWT-Food Science and Technology,2001,34(8):567-573.
[18] LI S Q,SHAH N P. Characterization,antioxidative and bifidogenic effects of polysaccharides from Pleurotus eryngii after heat treatments[J]. Food Chemistry,2016,197:240-249.
[19]苏红,李青连,魏磊,等.真菌多糖水提及化学辅助提取方法研究进展[J].微生物学通报,2010,37(3):426-432.SU H,LI Q L,WEI L,et al. Research progress in water extraction method and chemical assisted extraction method of fungal polysaccharides[J]. Microbiology China,2010,37(3):426-432.
[20] PEASURA N,LANHAKUNJIT N,KERDCHOECHUEN O,et al. Characteristics and antioxidant of Ulva intestinalis sulphated polysaccharides extracted with different solvents[J]. International Journal of Biological Macromolecules,2015,81(4):912-919.
[21]刘婷,周光明.多糖的提取和分析方法[J].化工时刊,2008,22(3):66-70.LIU T,ZHOU G M. Extraction and analytical method of polysaccharides[J]. Chemical Industry Times,2008,22(3):66-70.