桑木耳多糖提取工艺优化及其益生活性和抗氧化活性评价
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摘要
本文以云南昆明石林地区的桑木耳为原料,优化了桑木耳多糖水提工艺,并对多糖的益生活性和体外抗氧化活性进行分析。通过单因素试验结合响应面法获得了优化的桑木耳多糖水提工艺为:在料液比1:108(m/V),提取温度100℃,提取时间3.5 h时,单次提取率达6.96%,经10次重复提取,多糖总得率高达34.18%。经凝胶色谱柱测定桑木耳多糖的重均分子量为2.09×107 u。采用离子色谱对桑木耳多糖的单糖组成进行分析,发现其主要单糖组分为葡萄糖,木糖和甘露糖,摩尔质量比为1.18:0.65:2.88。红外光谱结果显示,该多糖含有β型吡喃糖苷键。益生活性和抗氧化活性研究表明,桑木耳多糖可有效促进两歧双歧杆菌和青春双歧杆菌的增殖,并具有良好的ABTS自由基和羟自由基清除能力。桑木耳多糖浓度为10 mg/m L时,对ABTS自由基、羟基自由基的清除率分别达到98.93%和71.28%。
The extraction process of of Mulberry fungus polysaccharide(MFP) sampled from Kunming, Yunnan Province, were optimized and the probiotic and antioxidant activities of the extracted polysaccharides were analyzed in this study. By single factor test combined with response surface methodology, the optimized water extraction conditions of MFP were obtained. When the extraction was performed with the solid-liquid ratio of 1:108(m/V) at 100℃ for 3.5 h, the polysaccharide yield was 6.96%. After 10 repeated extractions, the polysaccharide yield was as high as 34.18%. High performance gel permeation chromatography(HPGPC) analysis showed the average molecular weight of MFP was 2.09×107 u. The monosaccharide composition analyzed by ion chromatography showed that MFP mainly contained glucose, xylose and mannose, their molar mass ratio was 1.18: 0.65: 2.88. Fourier transform infrared(FT-IR) results showed that the polysaccharide contained β-glucopyranoside bonds. The experimental data of the effects of MFP on probiotics indicated that MFP had prebiotic activity that could effectively promote the proliferation of Bifidobacterium bifidum and Bifidobacterium adolescentis. Furthermore, the in vitro antioxidant activity experiment showed that MFP at the concentration of 10 mg/m L had strong ABTS·+ and ·OH radical scavenging ability and the scavenging rates were 98.93% and 71.28%, respectively.
The extraction process of of Mulberry fungus polysaccharide(MFP) sampled from Kunming, Yunnan Province, were optimized and the probiotic and antioxidant activities of the extracted polysaccharides were analyzed in this study. By single factor test combined with response surface methodology, the optimized water extraction conditions of MFP were obtained. When the extraction was performed with the solid-liquid ratio of 1:108(m/V) at 100℃ for 3.5 h, the polysaccharide yield was 6.96%. After 10 repeated extractions, the polysaccharide yield was as high as 34.18%. High performance gel permeation chromatography(HPGPC) analysis showed the average molecular weight of MFP was 2.09×107 u. The monosaccharide composition analyzed by ion chromatography showed that MFP mainly contained glucose, xylose and mannose, their molar mass ratio was 1.18: 0.65: 2.88. Fourier transform infrared(FT-IR) results showed that the polysaccharide contained β-glucopyranoside bonds. The experimental data of the effects of MFP on probiotics indicated that MFP had prebiotic activity that could effectively promote the proliferation of Bifidobacterium bifidum and Bifidobacterium adolescentis. Furthermore, the in vitro antioxidant activity experiment showed that MFP at the concentration of 10 mg/m L had strong ABTS·+ and ·OH radical scavenging ability and the scavenging rates were 98.93% and 71.28%, respectively.
引文
[1]Blinova M I,Yudintseva N M,Kalmykova N V,et al.Effect of melanins from black yeast fungi on proliferation and differentiation of cultivated human keratinocytes and fibroblasts[J].Cell Biology International,2003,27(2):135-146
[2]Vattem D A,Shetty K.Ellagic acid production and phenolic antioxidant activity in cranberry pomace(Vaccinium macrocarpon)mediated by Lentinus edodes using a solid-state system[J].Process Biochemistry,2003,39(3):367-379
[3]Yoon S J,Yu M A,Pyun Y R,et al.Nontoxic mushroom Auricularia auricula contains a polysaccharide with anticoagulant activity mediated by antithrombin[J].Thrombosis Research,2003,112(3):151-158
[4]Oke F,Aslim B.Protective effect of two edible mushrooms against oxidative cell damage and their phenolic composition[J].Food Chemistry,2011,128(3):613-619
[5]Adebayo-Tayo B C,Adegoke A A,Okoh A I,et al.Rationalizing some medicinal plants used in treatment of skin diseases[J].African Journal of Microbiology Research,2010,4(10):958-963
[6]Jin X.Bioactivities of water-soluble polysaccharides from fruit shell of Camellia oleifera Abel:Antitumor and antioxidant activities[J].Carbohydrate Polymers,2012,87(3):2198-2201
[7]Wang Y,Huang M,Sun R,et al.Extraction:Characterization of Ginseng fruits polysaccharide and its immune modulating activities in ratswith Lewis lung carcinoma[J].Carbohydrate Polymers,2015,127:215-221
[8]Murata Y,Shimamura T,Tagami T,et al.The skewing to Th1induced by lentinan is directed through the distinctive cytokine production by macrophages with elevated intracellular glutathione content[J].International Immunopharmacology,2002,2(5):673
[9]Kouakou K,Schepetkin I A,Yapi A,et al.Immunomodulatory activity of polysaccharides isolated from Alchornea cordifolia[J].Journal of Ethnopharmacology,2013,146:232-242
[10]Xu F F,Liao K S,Wu Y S,et al.Optimization,characterization,sulfation and antitumor activity of neutral polysaccharides from the fruit of Borojoa sorbilis cuter[J].Carbohydrate Polymers,2016,151:364-372
[11]Gao S,Lai C K,Cheng P C.Nondigestible carbohydrates isolated from medicinal mushroom sclerotia as novel prebiotics[J].International Journal of Medicinal Mushrooms,2009,11(1):1-8
[12]Snart J,Bibiloni R,Grayson T,et al.Supplementation of the diet with high-viscosity beta-glucan results in enrichment for lactobacilli in the rat cecum[J].Applied and Environmental Microbiology,2006,72(3):1925-1931
[13]郁志华.黄墩桑木耳生产技术[J].蚕桑通报,2011,42(2):61-62YU Zhi-hua.Production technology of mulberry fungus[J].Bulletin of Sericulture,2011,42(2):61-62
[14]叶良喜.桑木变废为宝育木耳[J].浙江林业,2007,11:27YE Liang-xi.Auricularia auricula cultivated with mulberry wood[J].Zhejiang Forestry,2007,11:27
[15]刘锐,陆建忠,王忠林,等.嘉兴市桑木耳营养成分检测[J].现代农业,2012,10:100-101LIU Rui,LU Jian-zhong,WANG Zhong-lin,et al.The nutrients detection of mulberry fungus from Jiaxing[J].Modern Agriculture,2012,10:100-101
[16]Dubois M,Gilles K A,Hamilton J K,et al.Colorimetric method for determination of sugars and related substances[J].Analytical Chemistry,1955,28(3):350-356
[17]Zhu X K,Zhang J Y,Nie S P,et al.Physicochemical properties and in vitro antioxidant activities ofpolysaccharide from Artocarpus heterophyllus Lam.Pulp[J].Carbohydrate Polymers,2017,155:354-361
[18]Li X C,Lin J,Gao Y X,et al.Antioxidant activity and mechanism of Rhizoma Cimicifugae[J].Chemistry Central Journal,2012,6:140
[19]Shao J,Yu J P,Hu M Z.Study on extraction and antioxidant activity of polysaccharides from Sarcandra glabra[J].Food Science,2007,28(11):283-286
[20]Zhang L N,Yang L Q,Qiong D,et al.Studies on molecular weights of polysaccharidesof Auricularia auricula-judae[J].Carbohydrate Research,1995,270:1-10
[21]Zeng W C,Zhang Z,Gao H,et al.Characterization of antioxidant polysaccharides from Auricularia auricular using microwave-assisted extraction[J].Carbohydrate Polymers,2012,89(2):694-700
[22]Wang Y,Li C,Liu P,et al.Physical characterization of exopolysaccharide produced by Lactobacillus plantarum KF5 isolated from Tibet Kefir[J].Carbohydrate Polymers,2010,82(3):895-903
[23]Melo M R,Feitosa J P,Freitas A L,et al.Isolation and characterization of soluble sulfated polysaccharide from the red seaweed Gracilaria cornea[J].Carbohydrate Polymers,2002,49(4):491-498
[24]Yu X H,Liu Y,Wu X L,et al.Isolation,purification,characterization and immunostimulatory activity of polysaccharides derived from American ginseng[J].Carbohydrate Polymers,2017,156:9-18
[25]Pawan G L.Molecular structure and function of food carbohydrates[J].Applied Science,1973,65-80
[26]Chen Y,Zhang H,Wang Y X,et al.Sulfated modification of the polysaccharides from Ganoderma atrum and their antioxidant and immunomodulating activities[J].Food Chemistry,2015,186(3):231-238
[27]Kadnikova I A,Costa R,Kalenik T K,et al.Chemical composition and nutritional value of the mushroom Auricularia auricula-judae[J].Journal of Food and Nutrition Research,2015,3(8):478-482
[2]Vattem D A,Shetty K.Ellagic acid production and phenolic antioxidant activity in cranberry pomace(Vaccinium macrocarpon)mediated by Lentinus edodes using a solid-state system[J].Process Biochemistry,2003,39(3):367-379
[3]Yoon S J,Yu M A,Pyun Y R,et al.Nontoxic mushroom Auricularia auricula contains a polysaccharide with anticoagulant activity mediated by antithrombin[J].Thrombosis Research,2003,112(3):151-158
[4]Oke F,Aslim B.Protective effect of two edible mushrooms against oxidative cell damage and their phenolic composition[J].Food Chemistry,2011,128(3):613-619
[5]Adebayo-Tayo B C,Adegoke A A,Okoh A I,et al.Rationalizing some medicinal plants used in treatment of skin diseases[J].African Journal of Microbiology Research,2010,4(10):958-963
[6]Jin X.Bioactivities of water-soluble polysaccharides from fruit shell of Camellia oleifera Abel:Antitumor and antioxidant activities[J].Carbohydrate Polymers,2012,87(3):2198-2201
[7]Wang Y,Huang M,Sun R,et al.Extraction:Characterization of Ginseng fruits polysaccharide and its immune modulating activities in ratswith Lewis lung carcinoma[J].Carbohydrate Polymers,2015,127:215-221
[8]Murata Y,Shimamura T,Tagami T,et al.The skewing to Th1induced by lentinan is directed through the distinctive cytokine production by macrophages with elevated intracellular glutathione content[J].International Immunopharmacology,2002,2(5):673
[9]Kouakou K,Schepetkin I A,Yapi A,et al.Immunomodulatory activity of polysaccharides isolated from Alchornea cordifolia[J].Journal of Ethnopharmacology,2013,146:232-242
[10]Xu F F,Liao K S,Wu Y S,et al.Optimization,characterization,sulfation and antitumor activity of neutral polysaccharides from the fruit of Borojoa sorbilis cuter[J].Carbohydrate Polymers,2016,151:364-372
[11]Gao S,Lai C K,Cheng P C.Nondigestible carbohydrates isolated from medicinal mushroom sclerotia as novel prebiotics[J].International Journal of Medicinal Mushrooms,2009,11(1):1-8
[12]Snart J,Bibiloni R,Grayson T,et al.Supplementation of the diet with high-viscosity beta-glucan results in enrichment for lactobacilli in the rat cecum[J].Applied and Environmental Microbiology,2006,72(3):1925-1931
[13]郁志华.黄墩桑木耳生产技术[J].蚕桑通报,2011,42(2):61-62YU Zhi-hua.Production technology of mulberry fungus[J].Bulletin of Sericulture,2011,42(2):61-62
[14]叶良喜.桑木变废为宝育木耳[J].浙江林业,2007,11:27YE Liang-xi.Auricularia auricula cultivated with mulberry wood[J].Zhejiang Forestry,2007,11:27
[15]刘锐,陆建忠,王忠林,等.嘉兴市桑木耳营养成分检测[J].现代农业,2012,10:100-101LIU Rui,LU Jian-zhong,WANG Zhong-lin,et al.The nutrients detection of mulberry fungus from Jiaxing[J].Modern Agriculture,2012,10:100-101
[16]Dubois M,Gilles K A,Hamilton J K,et al.Colorimetric method for determination of sugars and related substances[J].Analytical Chemistry,1955,28(3):350-356
[17]Zhu X K,Zhang J Y,Nie S P,et al.Physicochemical properties and in vitro antioxidant activities ofpolysaccharide from Artocarpus heterophyllus Lam.Pulp[J].Carbohydrate Polymers,2017,155:354-361
[18]Li X C,Lin J,Gao Y X,et al.Antioxidant activity and mechanism of Rhizoma Cimicifugae[J].Chemistry Central Journal,2012,6:140
[19]Shao J,Yu J P,Hu M Z.Study on extraction and antioxidant activity of polysaccharides from Sarcandra glabra[J].Food Science,2007,28(11):283-286
[20]Zhang L N,Yang L Q,Qiong D,et al.Studies on molecular weights of polysaccharidesof Auricularia auricula-judae[J].Carbohydrate Research,1995,270:1-10
[21]Zeng W C,Zhang Z,Gao H,et al.Characterization of antioxidant polysaccharides from Auricularia auricular using microwave-assisted extraction[J].Carbohydrate Polymers,2012,89(2):694-700
[22]Wang Y,Li C,Liu P,et al.Physical characterization of exopolysaccharide produced by Lactobacillus plantarum KF5 isolated from Tibet Kefir[J].Carbohydrate Polymers,2010,82(3):895-903
[23]Melo M R,Feitosa J P,Freitas A L,et al.Isolation and characterization of soluble sulfated polysaccharide from the red seaweed Gracilaria cornea[J].Carbohydrate Polymers,2002,49(4):491-498
[24]Yu X H,Liu Y,Wu X L,et al.Isolation,purification,characterization and immunostimulatory activity of polysaccharides derived from American ginseng[J].Carbohydrate Polymers,2017,156:9-18
[25]Pawan G L.Molecular structure and function of food carbohydrates[J].Applied Science,1973,65-80
[26]Chen Y,Zhang H,Wang Y X,et al.Sulfated modification of the polysaccharides from Ganoderma atrum and their antioxidant and immunomodulating activities[J].Food Chemistry,2015,186(3):231-238
[27]Kadnikova I A,Costa R,Kalenik T K,et al.Chemical composition and nutritional value of the mushroom Auricularia auricula-judae[J].Journal of Food and Nutrition Research,2015,3(8):478-482