香菇多酚氧化酶活性测定方法的改进
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摘要
对目前香菇多酚氧化酶活性的测定方法进行改进,通过离心消除蛋白质沉淀对反应的不利影响,以及其他因素对酶活力测定的影响,并对该方法进行了重复与再现试验。结果表明,香菇中多酚氧化酶的最佳测定条件为反应液预热温度40℃,添加质量分数20%(W/V)的三氯乙酸,取酶液1 m L,添加浓度0.3 mol/L的邻苯二酚,反应3 min后进行离心,于波长410 nm处测定吸光度。稳定性与再现性试验表明,此方法的重复试验和再现试验的相对标准偏差性均小于5%。
Common methods from literatures for the determination of polyphenol oxidase(PPO) activity in mushroom(Lentinus edodes(Berk.) sing) is improved. Adverse effect of precipitate is removed by centrifugation,at the same time,some factors are also studied,repetition and repeatability tests are conducted. The results show that the optimal conditions for the determination of PPO activity is as follows:preheating temperature of the reaction is 40 ℃,concentration of trichloroacetic acid is 20%(W/V),the crude enzyme is 1 m L,pyrocatechol is 0.3 mol/L after incubation for 3 min,the reaction liquid is centrifuged and measured the absorbance at 410 nm. Stability and repeatability rests show that the relative standard deviation of both tests is within 5%.
Common methods from literatures for the determination of polyphenol oxidase(PPO) activity in mushroom(Lentinus edodes(Berk.) sing) is improved. Adverse effect of precipitate is removed by centrifugation,at the same time,some factors are also studied,repetition and repeatability tests are conducted. The results show that the optimal conditions for the determination of PPO activity is as follows:preheating temperature of the reaction is 40 ℃,concentration of trichloroacetic acid is 20%(W/V),the crude enzyme is 1 m L,pyrocatechol is 0.3 mol/L after incubation for 3 min,the reaction liquid is centrifuged and measured the absorbance at 410 nm. Stability and repeatability rests show that the relative standard deviation of both tests is within 5%.
引文
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[14]莫祺晖,韦星明,李槐,等.三氯乙酸终止液对木瓜蛋白酶酶活测定的影响研究[J].饮料工业,2015(1):23-25.
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[16]吴进菊,陈红兵,高金燕,等.多种食用菌中多酚氧化酶活性的初步比较[J].食品与机械,2010(2):79-81.
[17]曲留柱.香蕉多酚氧化酶特性及其催化褐变防控的研究[D].北京:北京林业大学,2014.
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[19]巩慧玲,赵萍,袁惠君,等.马铃薯贮藏期间褐变强度与多酚氧化酶活性变化的研究[J].食品工业科技,2009(11):281-282,286.
[20]张洪,黄建韶.马铃薯中多酚氧化酶的酶学特性研究[J].食品工业科技,2002(4):39-41.
[21]孙月娥,张俊韬,王卫东.金针菇中多酚氧化酶活性测定及其护色研究[J].食品工业,2011(11):68-70.
[22]Jang M S,Sanada A,Ushio H,et al.Inhibitory effects of'Enokitake'mushroom extracts on polyphenol oxidase and prevention of apple browning[J].LWT-Food Science and Technology,2002,35(8):697-702.
[2]Gawlik-dziki U,Zlotek U,Swieca M.Characterization of polyphenol oxidase from butter lettuce(Lactuca sativa var.capitata L).[J].Food Chemistry,2008,107(1):129-135.
[3]周小玲,杨代明,李娜.小麦粉多酚氧化酶活性测定方法的改进[J].食品与机械,2015(6):84-87.
[4]譈NAL M譈,SENER A.Two-year comparison of the biochemical properties of polyphenol oxidase from Turkish Alyanak apricot(Prunus armenica L).[J].Food Chemistry,2016,190:741-747.
[5]汤凤霞,魏好程,曹禹.芒果多酚氧化酶的特性及抑制研究[J].食品科学,2006(12):156-160.
[6]苏适,黎莉,戴明,等.茶树菇中多酚氧化酶的活性研究[J].内江科技,2013(6):65-66.
[7]譈NAL M譈.Properties of polyphenol oxidase from Anamur banana(Musa cavendishi)i[J].Food Chemistry,2007,100(3):909-913.
[8]滕昆,张吉福,牛福星,等.脂肪酶活力测定中终止反应方法的比较分析[J].广西科学,2014(2):115-118.
[9]吴进菊.双孢蘑菇多酚氧化酶的研究[D].南昌:南昌大学,2010.
[10]耿中华,王乃馨.秀珍菇多酚氧化酶的特性研究[J].食品工业,2014(1):116-119.
[11]李君兰,李怡华,赵秋玲,等.鸡腿蘑多酚氧化酶特性研究[J].食品科学,2007(1):187-191.
[12]邵伟,乐超银,黄艺,等.蘑菇多酚氧化酶酶学特性初步研究[J].食用菌,2007(2):5-6.
[13]于新,冯彤,黄小丹.草菇多酚氧化酶及过氧化物酶活性的抑制研究[J].仲恺农业技术学院学报,2000(2):14-17,26.
[14]莫祺晖,韦星明,李槐,等.三氯乙酸终止液对木瓜蛋白酶酶活测定的影响研究[J].饮料工业,2015(1):23-25.
[15]刘雅嘉,李炜,衣杰荣.香菇多酚氧化酶酶学特性的研究[J].食品工业科技,2009(1):183-185,188.
[16]吴进菊,陈红兵,高金燕,等.多种食用菌中多酚氧化酶活性的初步比较[J].食品与机械,2010(2):79-81.
[17]曲留柱.香蕉多酚氧化酶特性及其催化褐变防控的研究[D].北京:北京林业大学,2014.
[18]Wuyts N,Waele D D,Swennen R.Extraction and partial characterization of polyphenol oxidase from banana(Musa acuminata Grande naine)roots[J].Plant Physiology and Biochemistry,2006,44(5):308-314.
[19]巩慧玲,赵萍,袁惠君,等.马铃薯贮藏期间褐变强度与多酚氧化酶活性变化的研究[J].食品工业科技,2009(11):281-282,286.
[20]张洪,黄建韶.马铃薯中多酚氧化酶的酶学特性研究[J].食品工业科技,2002(4):39-41.
[21]孙月娥,张俊韬,王卫东.金针菇中多酚氧化酶活性测定及其护色研究[J].食品工业,2011(11):68-70.
[22]Jang M S,Sanada A,Ushio H,et al.Inhibitory effects of'Enokitake'mushroom extracts on polyphenol oxidase and prevention of apple browning[J].LWT-Food Science and Technology,2002,35(8):697-702.