超声波及脱蛋白处理对猴头菌子实体多糖提取效果的影响
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摘要
本研究以猴头菌子实体为供试材料,在采用水提醇沉法进行多糖提取的过程中,系统研究了超声波和脱蛋白处理两个关键工艺环节对多糖提取效果的影响。研究结果表明:采用超声波辅助处理可提高猴头菌多糖提取率,降低多糖的纯度,使部分由蒸馏水洗脱获得的HEFP-α组分与全部0.1mol/LNaCl洗脱获得的HEFP-β组分经Sephacryl-S400洗脱后的曲线的小峰面积增加,也使主峰处的吸光值出现偏移。3种脱蛋白方法中,复合法对蛋白的除去效率最高,但多糖的保留率最低;亚铁氰化钾-乙酸锌法操作简便且蛋白除去率与多糖保留率均较高,但会使HEFP-β-2组分的含量和回收率降低;Sevage法蛋白脱除效率最低,多糖组分的保留率相对较高,且柱洗脱后发现经Sephacryl-S400凝胶柱分离后获得的主峰整体峰值较高,该方法较为适合多糖活性组分未知时的蛋白脱除。这一研究结果将会为该类大型真菌子实体的大分子生物活性物质的提取以及相关产品的开发提供重要的参考依据。
The effects of two key factors, ultrasound and deproteinization, on the quality of extracting polysaccharides(HEFPs) from Hericium erinaceus fruit-body were investigated using water extraction and alcohol precipitation method. The extracted samples were sequentially eluted by DEAE-52 ion exchange column and Sephacryl-S400 gel column. The result was that the extraction rate of HEFPs could be improved by ultrasound-assisted treatment, showing significant effect on the purity of HEFPs and increasing small peak areas of elution curves of partial HEFP-α component distilled by water and total HEFP-β component by0.1 mol/L NaCl, and presenting a shift in absorbance at the main peak eluted by Sephacryl-S400 gel column.Among three deproteinization methods, the method of combining ferrocyanide-zinc acetate with Sevage achieved the highest rate of protein removal, however, the polysaccharide retention rate was the lowest. The potassium ferrocyanide-zinc acetate method was simple and effective in protein removal and polysaccharide retention, but it could reduce the content and recovery rate of partial polysaccharide HEFP-β-2 component.The Sevage method had the lowest deproteinization efficiency and comparatively higher retention rate of HEFPs, showing highest main peak under elution of Sephacryl-S400 gel column, and it was suitable for protein removal when the bioactive components of polysaccharides were unknown. The data provide reference for extraction of macromolecular bioactive substances and further studies in exploring the related products of fruiting bodies of macrofungi.
The effects of two key factors, ultrasound and deproteinization, on the quality of extracting polysaccharides(HEFPs) from Hericium erinaceus fruit-body were investigated using water extraction and alcohol precipitation method. The extracted samples were sequentially eluted by DEAE-52 ion exchange column and Sephacryl-S400 gel column. The result was that the extraction rate of HEFPs could be improved by ultrasound-assisted treatment, showing significant effect on the purity of HEFPs and increasing small peak areas of elution curves of partial HEFP-α component distilled by water and total HEFP-β component by0.1 mol/L NaCl, and presenting a shift in absorbance at the main peak eluted by Sephacryl-S400 gel column.Among three deproteinization methods, the method of combining ferrocyanide-zinc acetate with Sevage achieved the highest rate of protein removal, however, the polysaccharide retention rate was the lowest. The potassium ferrocyanide-zinc acetate method was simple and effective in protein removal and polysaccharide retention, but it could reduce the content and recovery rate of partial polysaccharide HEFP-β-2 component.The Sevage method had the lowest deproteinization efficiency and comparatively higher retention rate of HEFPs, showing highest main peak under elution of Sephacryl-S400 gel column, and it was suitable for protein removal when the bioactive components of polysaccharides were unknown. The data provide reference for extraction of macromolecular bioactive substances and further studies in exploring the related products of fruiting bodies of macrofungi.
引文
Dai YC,2012.Pathogenic wood-decaying fungi on woody plants in China.Mycosystema,31(4):493-509(in Chinese)
Dai YC,Yang ZL,2008.A revised checklist of medicinal fungi in China.Mycosystema,27:801-824(in Chinese)
Dai YC,Zhou LW,Yang ZL,Wen HA,Bau T,Li TH,2010.Arevised checklist of edible fungi in China.Mycosystema,29:1-21(in Chinese)
Guo M,Ding GB,Guo SJ,Li ZY,Zhao LQ,Li K,Guo XR,2015.Isolation and antitumor efficacy evaluation of a polysaccharide from Nostoc commune Vauch.Food&Function,6:3035-3044
Guo SD,Cui YJ,Wang RZ,Wang RY,Wu WX,Ma T,2014.Separation,purification and primary reverse cholesterol transport study of Cordyceps militaris polysaccharide.China Journal of Chinese Materia Medica,39(17):3316(in Chinese)
He XR,Wang XX,Fang JC,Chang Y,Ning N,Guo H,Huang LH,Huang XQ,Zhao ZF,2017.Structures,biological activities,and industrial applications of the polysaccharides from Hericium erinaceus(lion’s mane)mushroom:a review.International Journal of Biological Macromolecules,87:228-237
Li G,Yu K,Li FS,Xu KP,Li J,He SJ,Cao SS,Tan GS,2014.Anticancer potential of Hericium erinaceus extracts against human gastrointestinal cancers.Journal of Ethnopharmacology,153:521-530
Li QZ,Wu D,Zhou S,Liu YF,Li ZP,Feng J,Yang Y,2016.Structure elucidation of a bioactive polysaccharide from fruiting bodies of Hericium erinaceus in different maturation stages.Carbohydrate Polymers,144:196-204
Lu R,Conrad P,Yacine H,2012.Antitumor activity of mushroom polysaccharides:a review.Food&Function,3:1118-1130
Sun YL,Dai HJ,Wen J,Zhang W,Zhao J,Tian L,Chen DH,2017.Optimization of enzymatic deproteinization of polysaccharides from cuttlefish muscle(Sepia lycidas)and its in vitro free radical scavenging potential.Food Science,38(18):251-258(in Chinese)
Tang W,Lin LH,Xie JH,Wang ZJ,Wang H,Dong YJ,Shen MY,Xie MY,2016.Effect of ultrasonic treatment on the physicochemical properties and antioxidant activities of polysaccharide from Cyclocarya paliurus.Carbohydrate Polymers,151:305-312
Wang LW,Ou YZ,Zhang BJ,Yang Q,Wang DJ,Yu YF,Zhang J,Ma Q,2017.Deproteinization and structural analysis of polysaccharides from Dendrobium officinale Kimura et Migo grown in Huoshan.Food Science,38(12):164-170(in Chinese)
Wang MX,Gao Y,Xu DD,Konishi T,Gao QP,2014.Hericium erinaceus(Yamabushitake):a unique resource for developing functional foods and medicines.Food&Function,5:3055-3064
Wang MX,Gao Y,Xu DD,Konishi T,Gao QP,2015.Apolysaccharide from cultured mycelium of Hericium erinaceus and its anti-chronic atrophic gastritis activity.International Journal of Biological Macromolecules,81:656-661
Wang XY,Yin JY,Nie SP,Xie MY,2018.Isolation,purification and physicochemical properties of polysaccharide from fruiting body of Hericium erinaceus and its effect on colonic health of mice.International Journal of Biological Macromolecules,107:1310-1319
Xu F,Wang CY,2014.Optimization of physical force-assisted alkaline extraction of rice bran protein.Food Science,35(20):11-16(in Chinese)
Xu YQ,Du MY,Yang L,Liu YS,Wei H,Zheng BH,Guo YY,Yang Y,2019.Effect of ultrasonic treatment on the properties and bioactivity of polysaccharide from blackcurrant fruit.Food Science,http://kns.cnki.net/kcms/detail/11.2206.ts.20181214.1353.036.html(in Chinese)
Xu YQ,Liu GJ,Yu ZY,Song XM,Li XG,Yang Y,Wang LB,Liu L,Dai J,2016.Purification,characterization and anti-glycation activity of a novel polysaccharide from black currant.Food Chemistry,199:694-701
Zhang AQ,Fu L,Xu M,2012.Structure of a water-soluble heteropolysaccharide from fruiting bodies of Hericium erinaceus.Carbohydrate Polymers,88(2):558-561
Zhang M,Cui SW,Cheung PCK,Wang Q,2007.Antitumor polysaccharides from mushrooms:a review on their isolation process,structural characteristics and antitumor activity.Trends in Food Science and Technology,18:4-19
戴玉成,2012.中国木本植物病原木材腐朽菌研究.菌物学报,31(4):493-509
戴玉成,杨祝良,2008.中国药用真菌名录及部分名称的修订.菌物学报,27:801-824
戴玉成,周丽伟,杨祝良,文华安,图力古尔,李泰辉,2010.中国食用菌名录.菌物学报,29:1-21
郭守东,崔英杰,王仁重,王仁远,武文雪,马腾,2014.蛹虫草多糖的分离纯化及促胆固醇逆向转运研究.中国中药杂志,39(17):3316-3320
孙玉林,戴宏杰,文菁,张卫,赵娟,田丽,陈道海,2017.拟目乌贼肌肉多糖酶法脱蛋白工艺优化及体外清除自由基能力.食品科学,38(18):251-258
王琳炜,欧阳臻,张碧娟,杨倩,王笃军,于一凡,张杰,马青,2017.霍山铁皮石斛多糖的脱蛋白工艺及结构分析.食品科学,38(12):164-170
许凤,王长远,2014.响应面法优化物理辅助碱法提取米糠蛋白工艺.食品科学,35(20):11-16
徐雅琴,杜明阳,杨露,刘育松,魏红,郑彬浩,郭莹莹,杨昱,2019.超声波处理对黑加仑果实多糖性质与生物活性的影响.食品科学,http://kns.cnki.net/kcms/detail/11.2206.ts.20181214.1353.036.html
Dai YC,Yang ZL,2008.A revised checklist of medicinal fungi in China.Mycosystema,27:801-824(in Chinese)
Dai YC,Zhou LW,Yang ZL,Wen HA,Bau T,Li TH,2010.Arevised checklist of edible fungi in China.Mycosystema,29:1-21(in Chinese)
Guo M,Ding GB,Guo SJ,Li ZY,Zhao LQ,Li K,Guo XR,2015.Isolation and antitumor efficacy evaluation of a polysaccharide from Nostoc commune Vauch.Food&Function,6:3035-3044
Guo SD,Cui YJ,Wang RZ,Wang RY,Wu WX,Ma T,2014.Separation,purification and primary reverse cholesterol transport study of Cordyceps militaris polysaccharide.China Journal of Chinese Materia Medica,39(17):3316(in Chinese)
He XR,Wang XX,Fang JC,Chang Y,Ning N,Guo H,Huang LH,Huang XQ,Zhao ZF,2017.Structures,biological activities,and industrial applications of the polysaccharides from Hericium erinaceus(lion’s mane)mushroom:a review.International Journal of Biological Macromolecules,87:228-237
Li G,Yu K,Li FS,Xu KP,Li J,He SJ,Cao SS,Tan GS,2014.Anticancer potential of Hericium erinaceus extracts against human gastrointestinal cancers.Journal of Ethnopharmacology,153:521-530
Li QZ,Wu D,Zhou S,Liu YF,Li ZP,Feng J,Yang Y,2016.Structure elucidation of a bioactive polysaccharide from fruiting bodies of Hericium erinaceus in different maturation stages.Carbohydrate Polymers,144:196-204
Lu R,Conrad P,Yacine H,2012.Antitumor activity of mushroom polysaccharides:a review.Food&Function,3:1118-1130
Sun YL,Dai HJ,Wen J,Zhang W,Zhao J,Tian L,Chen DH,2017.Optimization of enzymatic deproteinization of polysaccharides from cuttlefish muscle(Sepia lycidas)and its in vitro free radical scavenging potential.Food Science,38(18):251-258(in Chinese)
Tang W,Lin LH,Xie JH,Wang ZJ,Wang H,Dong YJ,Shen MY,Xie MY,2016.Effect of ultrasonic treatment on the physicochemical properties and antioxidant activities of polysaccharide from Cyclocarya paliurus.Carbohydrate Polymers,151:305-312
Wang LW,Ou YZ,Zhang BJ,Yang Q,Wang DJ,Yu YF,Zhang J,Ma Q,2017.Deproteinization and structural analysis of polysaccharides from Dendrobium officinale Kimura et Migo grown in Huoshan.Food Science,38(12):164-170(in Chinese)
Wang MX,Gao Y,Xu DD,Konishi T,Gao QP,2014.Hericium erinaceus(Yamabushitake):a unique resource for developing functional foods and medicines.Food&Function,5:3055-3064
Wang MX,Gao Y,Xu DD,Konishi T,Gao QP,2015.Apolysaccharide from cultured mycelium of Hericium erinaceus and its anti-chronic atrophic gastritis activity.International Journal of Biological Macromolecules,81:656-661
Wang XY,Yin JY,Nie SP,Xie MY,2018.Isolation,purification and physicochemical properties of polysaccharide from fruiting body of Hericium erinaceus and its effect on colonic health of mice.International Journal of Biological Macromolecules,107:1310-1319
Xu F,Wang CY,2014.Optimization of physical force-assisted alkaline extraction of rice bran protein.Food Science,35(20):11-16(in Chinese)
Xu YQ,Du MY,Yang L,Liu YS,Wei H,Zheng BH,Guo YY,Yang Y,2019.Effect of ultrasonic treatment on the properties and bioactivity of polysaccharide from blackcurrant fruit.Food Science,http://kns.cnki.net/kcms/detail/11.2206.ts.20181214.1353.036.html(in Chinese)
Xu YQ,Liu GJ,Yu ZY,Song XM,Li XG,Yang Y,Wang LB,Liu L,Dai J,2016.Purification,characterization and anti-glycation activity of a novel polysaccharide from black currant.Food Chemistry,199:694-701
Zhang AQ,Fu L,Xu M,2012.Structure of a water-soluble heteropolysaccharide from fruiting bodies of Hericium erinaceus.Carbohydrate Polymers,88(2):558-561
Zhang M,Cui SW,Cheung PCK,Wang Q,2007.Antitumor polysaccharides from mushrooms:a review on their isolation process,structural characteristics and antitumor activity.Trends in Food Science and Technology,18:4-19
戴玉成,2012.中国木本植物病原木材腐朽菌研究.菌物学报,31(4):493-509
戴玉成,杨祝良,2008.中国药用真菌名录及部分名称的修订.菌物学报,27:801-824
戴玉成,周丽伟,杨祝良,文华安,图力古尔,李泰辉,2010.中国食用菌名录.菌物学报,29:1-21
郭守东,崔英杰,王仁重,王仁远,武文雪,马腾,2014.蛹虫草多糖的分离纯化及促胆固醇逆向转运研究.中国中药杂志,39(17):3316-3320
孙玉林,戴宏杰,文菁,张卫,赵娟,田丽,陈道海,2017.拟目乌贼肌肉多糖酶法脱蛋白工艺优化及体外清除自由基能力.食品科学,38(18):251-258
王琳炜,欧阳臻,张碧娟,杨倩,王笃军,于一凡,张杰,马青,2017.霍山铁皮石斛多糖的脱蛋白工艺及结构分析.食品科学,38(12):164-170
许凤,王长远,2014.响应面法优化物理辅助碱法提取米糠蛋白工艺.食品科学,35(20):11-16
徐雅琴,杜明阳,杨露,刘育松,魏红,郑彬浩,郭莹莹,杨昱,2019.超声波处理对黑加仑果实多糖性质与生物活性的影响.食品科学,http://kns.cnki.net/kcms/detail/11.2206.ts.20181214.1353.036.html