桑黄菌丝体的液体发酵培养及多糖提取和抗氧化活性测定  

Liquid Fermentation of Phellinus igniarius Mycelium and Extraction and Antioxidant Activity Determination of Its Polysaccharide

作　　者：宋红志[1,2] 王方芹[1,2] 王俊[1,2] 盛晟[1,2] 吴福安[1,2] 

机构地区：[1]江苏科技大学,江苏镇江212003 [2]中国农业科学院蚕业研究所,江苏镇江212018

出　　处：《蚕业科学》2016年第4期700-710,共11页ACTA SERICOLOGICA SINICA

基　　金：江苏省科技支撑计划(农业)项目(No.BE2012365);现代农业产业技术体系建设专项(No.CARS-22);江苏省研究生科研创新计划项目(No.CXLX14_501)

摘　　要：为高效利用桑黄资源开发多糖类抗氧化剂,以菌丝体生物量为考核指标,对桑黄菌丝体的液体摇瓶发酵培养条件进行单因素试验,筛选出较佳的培养条件：以葡萄糖为碳源,大豆蛋白胨为氮源,发酵液p H为6,培养温度为30℃,摇瓶转速为140 r/min,接种量（体积分数）为4%。采用超声波辅助复合酶法提取桑黄多糖,复合酶由3%纤维素酶（酶活力为1.0×10~4U/g）、2%木瓜蛋白酶（1.5×10~4U/g）和1.5%果胶酶（4.0×10~4U/g）组成,通过单因素试验优化提取工艺条件为料液质量浓度20 g/L、超声波功率300 W、处理温度50℃、处理时间45 min,在此条件下多糖得率可达10.867%。采用分光光度法测定6个来源菌株培养桑黄菌丝提取粗多糖的抗氧化活性,结果显示6种桑黄粗多糖样品均表现出较强的体外抗氧化活性,其中韩国来源菌株、中国金寨来源菌株的桑黄多糖对ABTS和DPPH自由基的清除能力最强,具有进一步开发为天然抗氧化剂和自由基清除剂的潜力。In order to promote the development of polysaccharide antioxidant from Phellinus igniarius,single factor experiments were carried out to optimize the liquid fermentation conditions of P. igniarius mycelia by taking dry weight of mycelia as the evaluation index. The results showed that the best carbon source for the growth of P. igniarius mycelium was glucose,the optimal nitrogen source was soy peptone,the best fermentation p H was 6,the optimal growth temperature was 30 ℃,the best rotation rate and inoculation quantity（ volume fraction） was 140 r / min and 4% respectively. Furthermore,polysaccharide was extracted from P. igniarius by ultrasonic assisted digestion with composite enzymes which consisted of 3% cellulose（ 1. 0 × 10~4U / g）,2% papain protease（ 1. 5 × 10~4U / g） and 1. 5% pectinase（ 4. 0 × 10~4U / g）. The optimized extraction conditions obtained from single factor experiment was as following： the ratio of material to solvent was 20 g / L,ultrasonic power was 300 W,the extraction temperature was 50 ℃,and the extraction time was 45 min. The yield of polysaccharide was 10. 867% under this condition. The antioxidant activities of crude polysaccharide extracted from mycelia of 6 P. igniarius strains were evaluated by using spectrometric method. All samples showed high antioxidant activity,among which ABTS and DPPH free radical scavenging activities of polysaccharide extracted from mycelia of P. igniarius strains from Korean and Jinzhai,China were superior to others. The results showed that mycelium polysaccharide of P. igniarius has the potential for further development of natural antioxidant and free radical scavenger.

关 键 词：桑黄 发酵培养 多糖 超声波辅助复合酶法提取 抗氧化活性 

分 类 号：S567.39[农业科学—中草药栽培]