富硒香菇多糖发酵工艺的优化及其抗氧化活性
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摘要
本试验利用香菇作为载体,Na2Se O3为添加的硒源,优化富硒香菇多糖的发酵条件并研究其抗氧化活性。采用单因素试验考察了培养基p H、硒灭菌方式、硒添加时间、硒添加量和发酵时间对富硒香菇多糖的影响,并通过BoxBenhnken实验设计和响应面分析法确定最优发酵条件。在优化条件下得到富硒香菇多糖,并利用DPPH自由基清除能力研究其抗氧活性。结果表明:当培养基p H调至4.16,发酵至第162 h时,向发酵液中添加7.97 mg/L过滤除菌的Na2Se O3溶液,发酵11 d,富硒香菇胞内多糖提取量可达到71.54 mg/100 m L;当培养基p H调至4.01,发酵至第191.5 h时,向培养基中添加6.68 mg/L过滤除菌的Na2Se O3溶液,发酵11 d,富硒香菇胞外多糖提取量可达到853.96 mg/100 m L。DPPH自由基清除实验结果表明,抗氧化活性大小依次为富硒香菇胞内多糖>香菇胞内多糖>富硒香菇胞外多糖>香菇胞外多糖。硒的添加不仅促进香菇菌丝的生长及多糖的累积,同时也提高了其抗氧化活性,对DPPH有较好的清除作用。
In this experiment,Lentinula edods( L.edodes) was used as carrier,with Na2 Se O3 as the selenium source,to optimize the fermentation conditions of polysaccharide from Se-enriched L.edodes ant its antioxidant activity was studied. The effects of culture medium p H,sterling condition of selenium,selenium relenishing time,selenium concentration and incubation duration to polysaccharide form Se-enriched L.edodes were investigated by single factor experiment.The process parameters were optimized using a Box-Behnken experimental design coupled with response surface methodology. Polysaccharide from Se-enriched L.edodes was obtained under optimized conditions and its antioxidant activity was discussed by DPPH free radical scavening ability.The results showed that when the culture medium was p H4.16,7.97 mg/L Na2 Se O3 was added to the fermentation broth at 162 h,Na2 Se O3 solution was sterilized by filtration,the incubation duration was 11 d,the yield of crude intracelluar polysaccharide in Se-enriched L. edodes reached 71.54 mg/100 m L; when the culture medium p H was 4.01,6.68 mg/L Na2 Se O3 was added to the fermentation broth at 191.5 h,Na2 Se O3 solution was sterilized by filtration,the incubation duration was 11 d,the yield of crude extracellular polysaccharide in Se-enriched L. edodes reached 853.96 mg/100 m L. The results of DPPH free ladical scavening experiment showed that the antioxidative activities were as follows: intracellular polysaccharides in Se-enriched L. edodes > intracellular polysaccharides in L. edodes > extracellular polysaccharide in Se-enriched L. edodes >extracellular polysaccharide in L. edodes. Se-enriched not only increased the yield of polysaccharide form L. edodes,but also improved the antioxidant activity,better scavenging of the DPPH.
In this experiment,Lentinula edods( L.edodes) was used as carrier,with Na2 Se O3 as the selenium source,to optimize the fermentation conditions of polysaccharide from Se-enriched L.edodes ant its antioxidant activity was studied. The effects of culture medium p H,sterling condition of selenium,selenium relenishing time,selenium concentration and incubation duration to polysaccharide form Se-enriched L.edodes were investigated by single factor experiment.The process parameters were optimized using a Box-Behnken experimental design coupled with response surface methodology. Polysaccharide from Se-enriched L.edodes was obtained under optimized conditions and its antioxidant activity was discussed by DPPH free radical scavening ability.The results showed that when the culture medium was p H4.16,7.97 mg/L Na2 Se O3 was added to the fermentation broth at 162 h,Na2 Se O3 solution was sterilized by filtration,the incubation duration was 11 d,the yield of crude intracelluar polysaccharide in Se-enriched L. edodes reached 71.54 mg/100 m L; when the culture medium p H was 4.01,6.68 mg/L Na2 Se O3 was added to the fermentation broth at 191.5 h,Na2 Se O3 solution was sterilized by filtration,the incubation duration was 11 d,the yield of crude extracellular polysaccharide in Se-enriched L. edodes reached 853.96 mg/100 m L. The results of DPPH free ladical scavening experiment showed that the antioxidative activities were as follows: intracellular polysaccharides in Se-enriched L. edodes > intracellular polysaccharides in L. edodes > extracellular polysaccharide in Se-enriched L. edodes >extracellular polysaccharide in L. edodes. Se-enriched not only increased the yield of polysaccharide form L. edodes,but also improved the antioxidant activity,better scavenging of the DPPH.
引文
[1]高显钧,白裕兵,魏虹.我国富硒食品特色农业发展现状研究[J].中国食物与营养,2013,19(9):26-29.
[2]Lavu RVS,van de Wiele T,Pratti VL,et al.Selenium bioaccessibility in stomach,small intestine and colon:Comparison between pure Se compounds,Se-enriched food crops and food supplements[J].Food Chemistry,2016(197):382-387.
[3]戴玉成,杨祝良.中国药用真菌名录及部分名称的修订[J].菌物学报,2008,27(6):801-824.
[4]乔艳明,陈文强,徐晖.富硒香菇的研究现状及应用展望[J].陕西理工学院学报:自然科学版,2015,319(6):46-50.
[5]王丹,夏险,王革娇,等.微生物对硒的还原及其产物的应用研究进展--纪念硒发现200周年[J].微生物学通报,2017,44(7):1728-1735.
[6]乔艳明.富硒条件下香菇液体培养基优化及硒多糖提取工艺[D].汉中:陕西理工大学,2016.
[7]胡国元,游慧珍,董兰兰,等.茯苓菌丝体液体培养条件研究[J].武汉工程大学学报,2010,32(7):1-4.
[8]吴周斌,王锦锋,林占熺.真姬菇菌丝体对硒耐受性的研究[J].微生物学通报,2017,44(1):96-107.
[9]吕国英,张作法,潘慧娟,等.响应面分析法优化香菇多糖发酵培养基[J].菌物学报,2010,29(1):106-112.
[10]李玲飞.四种食用菌的富硒特性研究[D].杭州:浙江大学,2012.
[11]谢必峰,林琳,施巧琴,等.灵芝菌的深层培养及富硒特性[J].食品与发酵工业,1996(4):53-57
[12]高慧娟,杨霞霞,杜学基,等.荷叶离褶伞菌丝体富硒条件的研究[J].食品工业科技,2013,34(11):146-148.
[13]Yan Yaiuan,Li Xia,Wan Mianjie,et al.Effect of extraction methods on property and bioactivity of water-soluble polysaccharides from Amomum villosum[J].Carbohydrate Polymers,2015,117:632-635.
[14]郭雷,王淑军,刘玮炜.响应面法优化密蒙花总黄酮的超声提取工艺[J].食品科学,2012,33(4):13-17.
[15]周琪.富硒灵芝多糖的发酵及其抗氧化活性的初步研究[D].武汉:华中农业大学,2009.
[16]Eswayah AS,Smith TJ,Gardiner PHE.Microbial transformations of selenium species of relevance to bioremediation[J].Applied and Environmental Microbiology,2016,82(16):4848-4859.
[17]张国财,赵博,刘春延,等.响应面法优化超声波-微波协同提取富硒蛹虫草硒多糖工艺[J].食品科学,2016,37(12):33-39.
[18]刘全德,唐仕荣,王卫东,等.响应曲面法优化超声波-微波协同萃取生姜多糖工艺[J].食品科学,2010,31(18):124-128.
[19]陈默.发酵法制备羧甲基茯苓多糖的研究[D].武汉:武汉工程大学,2014.
[20]代向向.云芝生物转化无机硒生产富硒云芝糖肽的研究[D].上海:上海师范大学,2015.
[21]Mao Guanghua,Zou Ye,Feng Weiwei,et al.Extraction,preliminary characterization and antioxidant activity of Seenriched Maitake polysaccharide[J].Carbohydrate Polymers,2014,101:213-219.
[22]杨健华,秦惠娟,彭璐,等.草菇子实体提取物的抗氧化活性[J].食用菌学报,2017,24(2):55-58.
[23]Wang Y,Li Y,Liu Y,et al.Extraction,characterization and antioxidant activities of Se-enriched tea polysaccharides[J].International Journal of Biological Macromolecules,2015,77(6182):76-84.
[24]任瑞,马海东,刘斌,等.超声降解香菇多糖及其体外抗氧化活性的研究[J].安徽农业科学,2008,36(17):7061-7062.
[25]Chen Ruizhan,Jin Chengguang,Li Haiping,et al.Ultrahigh pressure extraction of polysaccharides form cordyceps militaris and evaluation of antioxidant activity[J].Separation and Purification Technology,2014,134:90-99.
[26]铁梅.食用菌中硒的形态分析[D].上海:华东师范大学,2006.
[27]刘涛,杨焱,冯涛,等.响应面法优化酶法水解香菇子实体中氨基酸的工艺[J].食用菌学报,2017,24(2):93-98.
[28]高慧娟,袁承玲,赵丽.响应面法优化富硒荷叶离褶伞菌丝体胞内多糖发酵条件[J].菌物学报,2016,35(1):86-93.
[29]Lavu RVS,van de Wiele T,Pratti VL,et al.Selenium bioaccessibility in stomach,small intestine and colon:Comparison between pure Se compounds,Se-enriched food crops and food supplements[J].Food Chemistry,2016,197:382-387.
[30]Li Shuang,Bian Fuling,Yue Ling,et al.Selenium-dependent antitumor immunomodulating activity of polysaccharides from roots of A.membranaceus[J].International Journal of Biological Macromolecules,2014,69:64-72.
[2]Lavu RVS,van de Wiele T,Pratti VL,et al.Selenium bioaccessibility in stomach,small intestine and colon:Comparison between pure Se compounds,Se-enriched food crops and food supplements[J].Food Chemistry,2016(197):382-387.
[3]戴玉成,杨祝良.中国药用真菌名录及部分名称的修订[J].菌物学报,2008,27(6):801-824.
[4]乔艳明,陈文强,徐晖.富硒香菇的研究现状及应用展望[J].陕西理工学院学报:自然科学版,2015,319(6):46-50.
[5]王丹,夏险,王革娇,等.微生物对硒的还原及其产物的应用研究进展--纪念硒发现200周年[J].微生物学通报,2017,44(7):1728-1735.
[6]乔艳明.富硒条件下香菇液体培养基优化及硒多糖提取工艺[D].汉中:陕西理工大学,2016.
[7]胡国元,游慧珍,董兰兰,等.茯苓菌丝体液体培养条件研究[J].武汉工程大学学报,2010,32(7):1-4.
[8]吴周斌,王锦锋,林占熺.真姬菇菌丝体对硒耐受性的研究[J].微生物学通报,2017,44(1):96-107.
[9]吕国英,张作法,潘慧娟,等.响应面分析法优化香菇多糖发酵培养基[J].菌物学报,2010,29(1):106-112.
[10]李玲飞.四种食用菌的富硒特性研究[D].杭州:浙江大学,2012.
[11]谢必峰,林琳,施巧琴,等.灵芝菌的深层培养及富硒特性[J].食品与发酵工业,1996(4):53-57
[12]高慧娟,杨霞霞,杜学基,等.荷叶离褶伞菌丝体富硒条件的研究[J].食品工业科技,2013,34(11):146-148.
[13]Yan Yaiuan,Li Xia,Wan Mianjie,et al.Effect of extraction methods on property and bioactivity of water-soluble polysaccharides from Amomum villosum[J].Carbohydrate Polymers,2015,117:632-635.
[14]郭雷,王淑军,刘玮炜.响应面法优化密蒙花总黄酮的超声提取工艺[J].食品科学,2012,33(4):13-17.
[15]周琪.富硒灵芝多糖的发酵及其抗氧化活性的初步研究[D].武汉:华中农业大学,2009.
[16]Eswayah AS,Smith TJ,Gardiner PHE.Microbial transformations of selenium species of relevance to bioremediation[J].Applied and Environmental Microbiology,2016,82(16):4848-4859.
[17]张国财,赵博,刘春延,等.响应面法优化超声波-微波协同提取富硒蛹虫草硒多糖工艺[J].食品科学,2016,37(12):33-39.
[18]刘全德,唐仕荣,王卫东,等.响应曲面法优化超声波-微波协同萃取生姜多糖工艺[J].食品科学,2010,31(18):124-128.
[19]陈默.发酵法制备羧甲基茯苓多糖的研究[D].武汉:武汉工程大学,2014.
[20]代向向.云芝生物转化无机硒生产富硒云芝糖肽的研究[D].上海:上海师范大学,2015.
[21]Mao Guanghua,Zou Ye,Feng Weiwei,et al.Extraction,preliminary characterization and antioxidant activity of Seenriched Maitake polysaccharide[J].Carbohydrate Polymers,2014,101:213-219.
[22]杨健华,秦惠娟,彭璐,等.草菇子实体提取物的抗氧化活性[J].食用菌学报,2017,24(2):55-58.
[23]Wang Y,Li Y,Liu Y,et al.Extraction,characterization and antioxidant activities of Se-enriched tea polysaccharides[J].International Journal of Biological Macromolecules,2015,77(6182):76-84.
[24]任瑞,马海东,刘斌,等.超声降解香菇多糖及其体外抗氧化活性的研究[J].安徽农业科学,2008,36(17):7061-7062.
[25]Chen Ruizhan,Jin Chengguang,Li Haiping,et al.Ultrahigh pressure extraction of polysaccharides form cordyceps militaris and evaluation of antioxidant activity[J].Separation and Purification Technology,2014,134:90-99.
[26]铁梅.食用菌中硒的形态分析[D].上海:华东师范大学,2006.
[27]刘涛,杨焱,冯涛,等.响应面法优化酶法水解香菇子实体中氨基酸的工艺[J].食用菌学报,2017,24(2):93-98.
[28]高慧娟,袁承玲,赵丽.响应面法优化富硒荷叶离褶伞菌丝体胞内多糖发酵条件[J].菌物学报,2016,35(1):86-93.
[29]Lavu RVS,van de Wiele T,Pratti VL,et al.Selenium bioaccessibility in stomach,small intestine and colon:Comparison between pure Se compounds,Se-enriched food crops and food supplements[J].Food Chemistry,2016,197:382-387.
[30]Li Shuang,Bian Fuling,Yue Ling,et al.Selenium-dependent antitumor immunomodulating activity of polysaccharides from roots of A.membranaceus[J].International Journal of Biological Macromolecules,2014,69:64-72.