南烛叶多酚氧化酶最适提取条件的研究
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摘要
[目的]深入探讨南烛叶多酚氧化酶(PPO)的酶学性质,对南烛叶PPO的最适提取条件进行研究。[方法]以南烛叶为原料,分别研究了交联聚乙烯吡咯烷酮(PVPP)添加含量、料液比、浸提时间和p H各因素对其提取的PPO酶活的影响。[结果]试验表明,适量南烛叶粉末,以料液比1∶15 g/m L,加入含9%PVPP(W/V)p H 7.6的柠檬酸-磷酸氢二钠缓冲液(0.1 mol/L),匀浆并于4℃下浸提9 h,过滤,滤液经冷冻离心(4℃,12 000 r/min,20 min)后得到酶活最高的PPO酶液(11 335 U/m L)。[结论]研究结果可为促进南烛叶的工业化应用提供参考。
[Objective] The aim was to explore the enzymatic properties of PPO from Vaccinium bracteatum Thunb leaf and research the optimum extraction conditions of PPO.[Method] Effects of PVPP content,solid-liquid ratio,extraction time,p H on PPO enzyme activity were studied respectively.[Result] The results showed that appropriate amount of V.bracteatum power was added into citric acid-phosphate buffer( 0.1 mol/L p H 7.6) containing 9% PVPP( W/V) with solid-liquid ratio 1∶ 15 g/m L,extracted at 4 ℃ for 9 h,through filtering,centrifuging at 4 ℃,12 000 r / min for 20 minutes,PPO with the highest enzyme activity( 11 335 U / m L) was obtained.[Conclusion]The study can provide reference for promoting industrialized application of V.bracteatum leaf.
[Objective] The aim was to explore the enzymatic properties of PPO from Vaccinium bracteatum Thunb leaf and research the optimum extraction conditions of PPO.[Method] Effects of PVPP content,solid-liquid ratio,extraction time,p H on PPO enzyme activity were studied respectively.[Result] The results showed that appropriate amount of V.bracteatum power was added into citric acid-phosphate buffer( 0.1 mol/L p H 7.6) containing 9% PVPP( W/V) with solid-liquid ratio 1∶ 15 g/m L,extracted at 4 ℃ for 9 h,through filtering,centrifuging at 4 ℃,12 000 r / min for 20 minutes,PPO with the highest enzyme activity( 11 335 U / m L) was obtained.[Conclusion]The study can provide reference for promoting industrialized application of V.bracteatum leaf.
引文
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[14]卢丽娜,卢婕,刘亮,等.奉化水蜜桃多酚氧化酶提取纯化工艺研究[J].湖北农业科学,2011,50(7):1451-1453.
[2]李立祥,吴红梅.提取方法对茶多酚氧化酶活性的影响[J].中国茶叶加工,2001(4):26-31.
[3]许雷.茶树多酚氧化酶的提取、分离纯化及其部分酶性质研究[D].武汉:华中农业大学,2014:9-18.
[4]徐广洲.乌饭树叶黑色素形成相关的酶与底物的初步研究[D].无锡:江南大学,2014:12-22.
[5]梁建荣.甘薯叶多酚氧化酶的分离纯化及部分与功能基团研究[D].重庆:西南大学,2011:32-40.
[6]WUYTS N,WAELE D D,SWENEN R.Extraction and partial characterization of polyphenol oxidase from banana(Musa acuminate Grand naine)roots[J].Plant physiology and biochemistry,2006,44:308-314.
[7]BENEDICTE L,VIRGINIE M L,TRISTAN R,et al.PVPP-polyphenol complexes:A mole-cular approach[J].Journal of agricultural and food chemistry,2006,54(12):4383-4389.
[8]SUN X H,ZHU K,ZHOU H M.Optimization of a novel backward extraction of defatted wheat germ protein from reverse micelles[J].Innovative food science and emerging technologies,2009,10:328-333.
[9]GAO Z J,HAN X H,XIAO X G.Puri cation and characterisation of polyphenol oxidase from red swiss chard[J].Food chemistry,2009,117:342-348.
[10]贺寅,王强,钟葵.响应面优化酶法提取龙眼多糖工艺[J].食品科学,2012,33(2):79-83.
[11]王璋.食品酶学[M].北京:中国轻工业出版社,2002.
[12]YORUK R,MARSHALL M R.Physicochemical properties and function of plant polyphenol oxidase:A review[J].Journal of food biochemistry,2003,27(5):361-422.
[13]刘琨,钱和,汪何雅.茶叶中多酚氧化酶提取的响应面优化[J].食品科技,2013,38(1):206-210.
[14]卢丽娜,卢婕,刘亮,等.奉化水蜜桃多酚氧化酶提取纯化工艺研究[J].湖北农业科学,2011,50(7):1451-1453.