为研究牛樟芝胞外多糖的抗氧化活性及其在胃肠系统中的消化特性,采用原位萃取发酵技术进行牛樟芝的液态发酵,并考察发酵萃取剂对多糖产量的影响;通过·O-2、·OH以及DPPH自由基清除率的测定评估胞外多糖的抗氧化性;构建体外模拟胃肠液和人源粪便的消化模型分析牛樟芝胞外多糖的体外消化特性。结果显示,油酸萃取发酵能够显著提高牛樟芝胞外多糖产量(236.42 mg/L)及抗氧化活性;利用简单醇沉就能够获得单一胞外多糖组分AC-2,其分子质量为23.07 kDa,主要由岩藻糖、半乳糖、葡萄糖和甘露糖组成,物质的量比为0.09∶1∶0.49∶0.27。此外,AC-2能够稳定通过胃肠液系统而不被消化降解并最终被肠道微生物利用;在人源粪便体外发酵过程中,AC-2显著降低肠道pH并提高短链脂肪酸(short chain fatty acids,SCFAs)含量,表明AC-2能够有效改善肠道环境。综上,AC-2作为一种小分子胞外多糖,能够稳定通过胃肠液系统,促进肠道微生物合成SCFAs,具有调节肠道健康的潜在活性。
[1] CHEN J F, TSAI Y T, LAI Y H, et al.Proteomic analysis of Antrodia cinnamomea-induced ER stress in liver cancer cells[J].Journal of Pharmaceutical and Biomedical Analysis, 2020, 187:113142.
[2] MANDAL E K, MAITY K, MAITY S, et al.Structural characterization of an immunoenhancing cytotoxic heteroglycan isolated from an edible mushroom Calocybe indica var.APK2[J].Carbohydrate Research, 2011, 346(14):2 237-2 243.
[3] ZHANG H N, MA H L, LIU W, et al.Ultrasound enhanced production and antioxidant activity of polysaccharides from mycelial fermentation of Phellinus igniarius[J].Carbohydrate Polymers, 2014, 113:380-387.
[4] MAO G H, REN Y, FENG W W, et al.Antitumor and immunomodulatory activity of a water-soluble polysaccharide from Grifola frondosa[J].Carbohydrate Polymers, 2015, 134:406-412.
[5] WANG Z Q, ZHU C X, DAI A R, et al.Chemical characterization and antioxidant properties of cell wall polysaccharides from Antrodia cinnamomea mycelia[J].Food Bioscience, 2021, 41:100932.
[6] LEE M H, CHAO C H, HSU Y C, et al.Production, characterization, and functions of sulfated polysaccharides from zinc sulfate enriched cultivation of Antrodia cinnamomea[J].International Journal of Biological Macromolecules, 2020, 159:1 013-1 021.
[7] ZHANG Y T, WANG Z, LI D Y, et al.A polysaccharide from Antrodia cinnamomea mycelia exerts antitumor activity through blocking of TOP1/TDP1-mediated DNA repair pathway[J].International Journal of Biological Macromolecules, 2018, 120:1 551-1 560.
[8] LIU Y T, LI Y W, KE Y, et al.In vitro saliva-gastrointestinal digestion and fecal fermentation of Oudemansiella radicata polysaccharides reveal its digestion profile and effect on the modulation of the gut microbiota[J].Carbohydrate Polymers, 2021, 251:117041.
[9] LI X, XIE Q T, HUANG S M, et al.Digestion & fermentation characteristics of sulfated polysaccharides from Gracilaria chouae using two extraction methods in vitro and in vivo[J].Food Research International, 2021, 145:110406.
[10] LUO Z, WANG L, ZHOU P, et al.Effect of in vitro simulated gastrointestinal digestion on structural characteristics and anti-proliferative activities of the polysaccharides from the shells of Juglans regia L[J].Food and Chemical Toxicology, 2021, 150:112100.
[11] CAO Z H, GUO Y, LIU Z H, et al.Ultrasonic enzyme-assisted extraction of comfrey (Symphytum officinale L.) polysaccharides and their digestion and fermentation behaviors in vitro[J].Process Biochemistry, 2022, 112:98-111.
[12] LIU X F, XIA Y J, ZHANG Y, et al.Enhancement of antroquinonol production via the overexpression of 4-hydroxybenzoate polyprenyltransferase biosynthesis-related genes in Antrodia cinnamomea[J].Phytochemistry, 2021, 184:112677.
[13] 郑若欣, 易啸, 罗爽, 等.醇沉-苯酚硫酸法测定桑葚酒中多糖含量[J].现代食品, 2021(9):154-158.
ZHENG R X, YI X, LUO S, et al.Determination of polysaccharide in mulberry wine by ethanol precipitation-phenol sulfuric acid method[J].Modern Food, 2021(9):154-158.
[14] 赵英永, 戴云, 崔秀明, 等.考马斯亮蓝G-250染色法测定草乌中可溶性蛋白质含量[J].云南民族大学学报(自然科学版), 2006, 15(3):235-237.
ZHAO Y Y, DAI Y, CUI X M, et al.Determination of protein contents of Radix Aconiti kusnezoffii using coomassie brillant blue G-250 dye binding[J].Journal of Yunnan Nationalities University (Natural Sciences Edition), 2006, 15(3):235-237.
[15] 杨鸿玲, 李淑芬, 贾冬冬.桑叶多糖中糖醛酸和中性糖含量测定方法的建立[J].药物分析杂志, 2009, 29(2):301-304.
YANG H L, LI S F, JIA D D.A method for determination of the neutral sugars and uronic acids in mulberry leaves[J].Chinese Journal of Pharmaceutical Analysis, 2009, 29(2):301-304.
[16] MARKLUND S, MARKLUND G.Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase[J].European Journal of Biochemistry, 1974, 47(3):469-474.
[17] FAN M H, SUN X, QIAN Y L, et al.Effects of metal ions in tea polysaccharides on their in vitro antioxidant activity and hypoglycemic activity[J].International Journal of Biological Macromolecules, 2018, 113:418-426.
[18] FENG K, CHEN W, SUN L W, et al.Optimization extraction, preliminary characterization and antioxidant activity in vitro of polysaccharides from Stachys sieboldii Miq.tubers[J].Carbohydrate Polymers, 2015, 125:45-52.
[19] YUAN Y Q, LI C, ZHENG Q W, et al.Effect of simulated gastrointestinal digestion in vitro on the antioxidant activity, molecular weight and microstructure of polysaccharides from a tropical sea cucumber (Holothuria leucospilota)[J].Food Hydrocolloids, 2019, 89:735-741.
[20] 高文军, 李卫红, 王喜明, 等.3, 5-二硝基水杨酸法测定蔓菁中还原糖和总糖含量[J].中国药业, 2020, 29(9):113-116.
GAO W J, LI W H, WANG X M, et al.Determination of reducing sugar and total sugar in turnip by 3, 5-dinitrosalicylic acid colorimetry[J].China Pharmaceuticals, 2020, 29(9):113-116.
[21] 张冠亚. 铁皮石斛多糖在模拟消化、酵解体系中的代谢特点及其改善肠道功能的研究[D].南昌:南昌大学, 2015.
ZHANG G Y.Research on metabolic characteristics of Dendrobium officinale polysaccharides in simulating digestion and fermentation system and its effect on the improvement of intestinal function[D].Nanchang:Nanchang University, 2015.
[22] 杨彩云, 刘晓凤, 王光强, 等.响应面法优化提高桑黄YC-1菌发酵法生产总三萜产率的研究[J].食品发酵与科技, 2021, 57(4):1-7;31.
YANG C Y, LIU X F, WANG G Q, et al.Optimization and improvement of total triterpenes yield of Phellinus sp. YC-1 by fermentation with response surface methodology[J].Food and Fermentation Sciences & Technology, 2021, 57(4):1-7;31.
[23] YI J P, LI X, WANG S, et al.Steam explosion pretreatment of Achyranthis bidentatae radix:Modified polysaccharide and its antioxidant activities[J].Food Chemistry, 2022, 375:131746.
[24] 吴雅清, 冷小鹏.多糖体外抗氧化作用及其影响因素[J].广州化工, 2018, 46(4):4-9;16.
WU Y Q, LENG X P.Antioxidant activity and influencing factors of polysaccharides in vitro[J].Guangzhou Chemical Industry, 2018, 46(4):4-9;16.
[25] LI J J, PANG B, YAN X M, et al.Prebiotic properties of different polysaccharide fractions from Artemisia sphaerocephala Krasch seeds evaluated by simulated digestion and in vitro fermentation by human fecal microbiota[J].International Journal of Biological Macromolecules, 2020, 162:414-424.
[26] WANG M J, CHEN G J, CHEN D, et al.Purified fraction of polysaccharides from Fuzhuan brick tea modulates the composition and metabolism of gut microbiota in anaerobic fermentation in vitro[J].International Journal of Biological Macromolecules, 2019, 140:858-870.