酶法提取苦荞D-手性肌醇及体外降糖活性  

Enzymatic Extraction of D-chiro-inositol in Tartary Buckwheat and Its in Vitro Hypoglycemic Activity

作　　者：田晓艳 白晓琳[1] 王振宇[2] 张大千 樊梓鸾[1,3] TIAN Xiaoyan;BAI Xiaolin;WANG Zhenyu;ZHANG Daqian;FAN Ziluan(College of Forestry,Northeast Forestry University,Harbin 150040,China;College of Food Science,Harbin Institute of Technology,Harbin 150090,China;Heilongjiang Provincial Key Laboratory of Forest Food Resources Utilization,Harbin 150040,China)

机构地区：[1]东北林业大学林学院,黑龙江哈尔滨150040 [2]哈尔滨工业大学食品学院,黑龙江哈尔滨150090 [3]黑龙江省森林食品资源利用重点实验室,黑龙江哈尔滨150040

出　　处：《现代食品科技》2023年第5期32-40,共9页Modern Food Science and Technology

基　　金：国家自然科学基金项目(31170510);黑龙江省大学生创新训练项目(202110225278);黑龙江省科学自然基金项目(JJ2020LH1480);中央高校科学前沿与交叉学科创新基金项目(2572019BA09);黑龙江省高等教育教学改革项目(SJGY20200018);中国博士后科学基金(2016M600239)。

摘　　要：该实验以苦荞籽粒为研究对象,使用酶辅助超声法提取D-手性肌醇(D-Chiro Inositol,DCI)。考察了pH值、酶解温度、酶解时间、加酶量、料液比、乙醇体积分数对DCI浓度的影响,并在此基础上,采用响应面试验设计优化了DCI最佳提取工艺。结果显示:酶解最佳pH值为6,酶解温度为36.4℃,酶解时间为0.5 h,加酶量为2.1%,料液比为1:10,乙醇体积分数为48%时,DCI浓度为5.87 mmol/L。进一步利用α-淀粉酶及α-葡萄糖苷酶两项指标评价提取物纯化前后的体外降糖能力。采用活性炭、固相萃取离子小柱纯化DCI,结果显示阴离子(SAX)固相萃取小柱纯化效果最好,能够将纯度提高80%。苦荞中DCI对α-葡萄糖苷酶的半抑制质量浓度(50%Inhibitory Concentration,IC50)降低,IC50值由0.68 mg/mL降到0.47 mg/mL,表明纯化效果显著。采用α-半乳糖苷酶辅助超声法可以有效的获得高浓度DCI,并证实纯化后的DCI具有较好的体外降糖活性。通过该项目的研究为苦荞的综合开发利用提供理论参考。Using Tartary buckwheat grains as the research object,D-chiro-inositol(DCI)was extracted via an enzyme-assisted ultrasonic method.The effects of pH,enzymatic hydrolysis temperature,enzymatic hydrolysis time,enzyme concentration,solid-liquid ratio,and ethanol concentration on DCI concentration were investigated,and the extraction process of DCI was optimized using response surface methodology.The optimal enzymatic hydrolysis pH was 6,enzymatic hydrolysis temperature was 36.4,enzymatic hydrolysis℃time was 0.5 h,enzyme concentration was 2.1%,solid-liquid ratio was 1:10,and ethanol concentration was 48%;as such,DCI concentration measured under these conditions was 5.87 mmol/L.Further,the in vitro hypoglycemic activity of the extract before and after purification was evaluated usingα-amylase andα-glucosidase.DCI was purified with activate carbon and a solid-phase extraction cartridge.Anion(SAX)solid-phase extraction cartridge produced the best purification effect,increasing the purity by 80%.The 50%inhibitory concentration(IC50)of DCI onα-glucosidase in Tartary buckwheat decreased from 0.675 to 0.472 mg/mL,indicating a significant purification effect.The proposedα-galactosidase-assisted ultrasound method effectively produced a high concentration of DCI,and the purified DCI showed excellent in vitro hypoglycemic activity.These findings provide a theoretical reference for the comprehensive development and utilization of Tartary buckwheat.

关 键 词：D-手性肌醇(DCI) Α-半乳糖苷酶 降糖活性 苦荞 

分 类 号：TS210.4[轻工技术与工程—粮食、油脂及植物蛋白工程]