鸡腿菇多糖的液态发酵工艺优化及抗氧化活性
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摘要
在单因素试验基础上,采用Box-Behnken试验设计对鸡腿菇多糖液态发酵工艺进行响应面法优化,并对鸡腿菇多糖的抗氧化性进行研究。结果表明,最优发酵工艺为玉米粉3%,蔗糖3%,黄豆饼粉0.2%,酵母膏0.2%,装液量90 mL/250 mL,转速160 r/min,25℃条件下发酵6 d。在此最佳条件下,鸡腿菇菌丝体生物量为3.2 g/100 mL,胞外多糖产量为2.9 g/L。抗氧化试验结果表明,质量浓度为2 mg/m L鸡腿菇胞内多糖(IPS)和胞外多糖(EPS)对DPPH·的清除率分别达到55.1%和47.4%;质量浓度为8 mg/m L鸡腿菇胞内菌丝多糖和胞外多糖对·OH的清除率分别达到52.2%和44.1%、对NO·的清除率分别达到65.1%和48.1%、对O_2-·的清除率分别达到27.2%和38.2%;还原力(OD_(700) nm值)分别达到0.52和0.78,对亚铁离子(Fe~(2+))诱发的卵黄脂蛋白脂质过氧化抑制率分别为25.2%和37.1%。试验证明鸡腿菇多糖具有良好的抗氧化活性。
Based on the single factor experiments, the liquid-state fermentation technology of Coprinus comatus was optimized using Box-Behnken experimental design, and the antioxidant activity of C. comatus polysaccharides was studied. The results showed that the optimal fermentation technology was corn flour 3%, sucrose 3%, soybean powder 0.2%, yeast extract 0.2%, liquid loading volume 90 ml/250 ml, rotation speed 160 r/min, and fermentation time 6 d at 25 ℃. Under the optimal condition, the mycelial biomass of C. comatus was 3.2 g/100 ml, and the extracellular polysaccharide yield was 2.9 g/L. The antioxidant activity results showed that the DPPH scavenging rate of 2 mg/ml C. comatus intracellular polysaccharides(IPS)and extracellular polysaccharides(EPS) could reach 55.1% and 47.4%, respectively. When the IPS and EPS concentration was 8 mg/ml, the ·OH scavenging rate was 52.2% and 44.1%, respectively; the NO·scavenging rate was 65.1% and 48.1%, respectively; and the O_2-·scavenging rate was 27.2% and 38.2%, respectively. The reducing power(OD_(700) nmvalue) reached 0.52 and 0.78, and the inhibition rate of ferrous ion(Fe~(2+))-induced yolk lipoprotein lipid peroxidation were 25.2% and 37.1%. The test proved that the polysaccharide of C. comatus had good antioxidant activity.
Based on the single factor experiments, the liquid-state fermentation technology of Coprinus comatus was optimized using Box-Behnken experimental design, and the antioxidant activity of C. comatus polysaccharides was studied. The results showed that the optimal fermentation technology was corn flour 3%, sucrose 3%, soybean powder 0.2%, yeast extract 0.2%, liquid loading volume 90 ml/250 ml, rotation speed 160 r/min, and fermentation time 6 d at 25 ℃. Under the optimal condition, the mycelial biomass of C. comatus was 3.2 g/100 ml, and the extracellular polysaccharide yield was 2.9 g/L. The antioxidant activity results showed that the DPPH scavenging rate of 2 mg/ml C. comatus intracellular polysaccharides(IPS)and extracellular polysaccharides(EPS) could reach 55.1% and 47.4%, respectively. When the IPS and EPS concentration was 8 mg/ml, the ·OH scavenging rate was 52.2% and 44.1%, respectively; the NO·scavenging rate was 65.1% and 48.1%, respectively; and the O_2-·scavenging rate was 27.2% and 38.2%, respectively. The reducing power(OD_(700) nmvalue) reached 0.52 and 0.78, and the inhibition rate of ferrous ion(Fe~(2+))-induced yolk lipoprotein lipid peroxidation were 25.2% and 37.1%. The test proved that the polysaccharide of C. comatus had good antioxidant activity.
引文
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[3]余杰,崔鹏举,卓健勇.响应面分析方法在鸡腿菇多糖提取中的应用[J].食品科学,2009,30(8):93-96.
[4]LI B,DOBRUCHOWSKA J M,GERWIG G J,et al.Structural investigation of water-soluble polysaccharides extracted from the fruit bodies of Coprinus comatus[J].Carbohyd Polym,2013,91(1):314-321.
[5]王岩,关洪全,马贤德.毛头鬼伞多糖对免疫抑制小鼠体液免疫功能的影响[J].吉林中医药,2010,30(3):260-261.
[6]赵源.毛头鬼伞多糖制备工艺优化及高分子量多糖结构和免疫活性研究[D].南京:南京农业大学,2012.
[7]LIU Y,ZHAO Y,YANG Y,et al.Structural characteristics and hypoglycemic activity of polysaccharides from Coprinus comatus[J].Bioact Carbohydr Diet Fibre,2013,2(2):164-169.
[8]吴艳兵.毛头鬼伞(Coprinus comatus)多糖的分离纯化及其抗烟草花叶病毒(TMV)作用机制[D].福州:福建农林大学,2007.
[9]ZHAO H J,LI H P,LAI Q Q,et al.Antioxidant and hepatoprotective activities of modified polysaccharides from Coprinus comatus in mice with alcohol-induced liver injury[J].Int J Biol Macromol,2019,127(4):476-485.
[10]金卫根,吴瑞娟.毛头鬼伞液体培养条件试验[J].食用菌,2008,30(4):12-14.
[11]李巍巍,刘晓兰,时晰,等.鸡腿菇产溶栓酶液体发酵条件优化[J].工业微生物,2009,39(2):49-54.
[12]凡军民.毛头鬼伞菌丝体多糖的分离、纯化和化学结构鉴定及生物活性研究[D].南京:南京农业大学,2006.
[13]陆武祥,王东华,王秀英,等.5种食用菌液体发酵菌丝抗氧化活性分析比较[J].食品与发酵工业,2013,39(7):124-127.
[14]TSAI S Y,TSAI H L,MAU J L.Antioxidant properties of Coprinus comatus[J].J Food Biochem,2010,33(3):368-389.
[15]CHEN H,XU X,ZHU Y.Optimization of hydroxyl radical scavenging activity of exopolysaccharides from Inonotus obliquus in submerged fermentation using response surface methodology[J].J Microbiol Biotech,2010,20(4):835-843.
[16]金萍.鸡腿菇富锌液体发酵条件的研究[J].苏州科技大学学报:自然科学版,2014(2):47-51.
[17]胡梅.鸡腿蘑深层发酵技术及其多糖分离纯化的研究[D].武汉:华中农业大学,2006.
[18]韩秋菊.鸡腿菇多糖提取工艺条件的研究[J].陕西农业科学,2012,58(3):63-65.
[19]孙汉巨.菜籽多糖的分离、纯化及特性研究[D].合肥:合肥工业大学,2009.
[20]王金玺,顾林,孔凡伟,等.鸡腿菇粗多糖的体外抗氧化性[J].食品科学,2012,33(13):79-82.
[21]PRICTO P,PINEDA M,AGUILAR M.Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex:specific application to the determination of vitamin E[J].Anal Biochem,1999,269(2):337-341.