| 摘要: |
| [目的]优化猪苓发酵菌丝胞内多糖的提取工艺,以提高猪苓胞内多糖浸提的得率.[方法]通过单因素试验、正交试验和对比试验确定猪苓发酵菌丝胞内多糖的浸提方法、辅助浸提方法和除蛋白方法.[结果]与水浸提相比,弱碱性溶剂浸提猪苓菌丝胞内多糖的效果更好,具体工艺参数为:按料液比1:40添加40倍体积pH=11的NaOH,90℃浸提4次,每次60 min;酶解辅助法和超声波辅助法可有效提高猪苓发酵菌丝胞内多糖的浸提得率,其中酶解辅助浸提的效果更好,其最佳工艺参数为:复合酶配比为m(纤维素酶):m(果胶酶)=1:1,复合酶用量为10 mg/g,酶解80 min;蛋白酶和Sevag配合的除蛋白方法,是去除猪苓胞内蛋白的最佳方法.[结论]猪苓发酵菌丝胞内多糖的最适提取工艺流程为:猪苓菌丝干粉-添加10 mg/g复合酶[m(纤维素酶):m(果胶酶)=1:1]和40倍体积蒸馏水-50℃酶解80 min-NaOH溶液调pH=11,90℃浸提4次,每次60 min-浸提液合并、浓缩-HCl溶液调pH=6.5-蛋白酶酶解1 h-85℃酶灭活-Sevag除蛋白至无蛋白检出-浓缩-醇沉-干燥. |
| 关键词: 猪苓 胞内多糖 提取工艺 辅助浸提 蛋白酶 |
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| 基金项目:教育部新世纪优秀人才支持计划项目；西北农林科技大学拔尖人才支持计划项目 |
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| Extraction methods of polysaccharides from the fermentation mycelium of Polyporus umbellatus |
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| Abstract: |
| 【Objective】 The study was done to maximize the yield of intracellular polysaccharides (IPS) extracted from Polyporus umbellatus.【Method】 The solvent extraction,assistant treatments and the methods of protein removing were studied by single-factor tests,orthogonal experiments and comparison tests.【Result】 The results indicated that the optimal extraction conditions of IPS were pH=11 solvent,temperature of 90 ℃,4 times extraction,and 60 min each time respectively.And the extraction rates of IPS were enhanced by enzyme treatment and ultrasonic-assisted treatment before hot-water extraction,and the optimal conditions of enzyme treatment were m(Cellulase)∶m(Pectinase)= 1∶1,enzyme dosage of 10 mg/g,and time of 80 min.Furthermore,this study also indicated that the protein of crude polysaccharide which was treated with proteinase was easy to be removed by Sevag method.【Conclusion】 The most suitable extraction process of polysaccharides from Polyporus umbellatus was gained from this study,that is 10 mg/g compound enzyme〔m(Cellulase)∶m(Pectinase)= 1∶1〕treaded dry mycelium for 80 min in 40 (m/V) distilled water,temperature at 50 ℃;then adjusted pH to 11 with NaOH,extracted at 90 ℃ for 4 times,60 min each time;then adjusted pH to 6.5 with HCl,proteinase treated for 1 h,inactivated enzyme at 85 ℃,removed protein with the method of Sevag;finally,after concentrated and alcohol deposit, polysaccharide of P.umbellatus could be gained. |
| Key words: Polyporus umbellatus intracellular polysaccharides(IPS) extraction process assisted extraction proteinase |
