60 Hz低频电磁场对α-淀粉酶催化效率的影响及作用机理
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摘要
为了研究低频电磁场对酶促反应的影响及其作用机理,本文以α-淀粉酶催化水解玉米淀粉为酶促反应模型,使用60 Hz低频电磁场(LF-EMF)进行持续辐射处理,通过分析低频电磁场辐射对酶促淀粉水解反应效率的影响,以及低频电磁场辐射对底物结构、酶结构、及溶液体系传质等方面的影响,初步探讨了低频电磁场对酶促反应影响的作用机理。结果表明:低频电磁场可促进α-淀粉酶催化的淀粉水解反应,反应10 h时,产水解糖达14.39 mg/mL,比对照实验(12.41 mg/mL)提高了15.90%(p<0.05);低频电磁场对玉米淀粉的形貌及结晶度无显著影响(p>0.05);未发现低频电磁场引起α-淀粉酶一级结构的明显变化,也未发现低频电磁场辐射导致酶的二级结构/三级结构不可逆的改变(p>0.05);低频电磁场可降低α-淀粉酶催化反应体系的最适离子强度(0.0947 mol/L),并降低高离子强度对酶促反应的抑制作用;氯化钠的扩散实验表明低频电磁场可促进溶液体系的传质,其中,离子扩散的增强因子为1.23,表观膜厚度由1.26 cm减少到1.08 cm,降低了14.29%(p<0.05)。LF-EMF辐射提高了α-淀粉酶催化玉米淀粉水解的反应效率,原因之一是强化了反应传质,但不排除其影响α-淀粉酶的构象进而提高酶活性的可能性。
To investigate the effect and mechanism of low frequency electromagnetic field(LF-EMF)on enzymatic reaction,in this paper,the hydrolysis of corn starch catalyzed by α-amylase was used as a model reaction,treatment on the reaction with 60 Hz LF-EMF was carried out. Based on the analyzing of the LF-EMF's influence on the reaction efficiency of enzymatic starch hydrolysis,and its influence on the substrate structure,the enzyme structure and the mass transfer in solution system,the mechanism of the LF-EMF's influence on the enzymatic reaction was discussed preliminarily. The results showed that LF-EMF radiation promoted the hydrolysis of starch catalyzed by α-amylase,14.39 mg/mL hydrolyzed sugar was produced in 10 h reaction,which was 15.90% higher than the control(12.41 mg/mL)(p<0.05);Neither remarkable change on the primary structure of α-amylase,nor irreversible change on the secondary structure/tertiary structure resulted from LF-EMF radiation was observed(p>0.05). The LF-EMF could reduce the optimum ionic strength(which was 0.0947 mol/L)of the α-amylase's reaction system,and weaken the inhibition of high ionic strength on the enzymatic reaction. The sodium chloride diffusion experiment indicated that LF-EMF could promote mass transfer in solution system,the enhancement factor of ion diffusion was 1.23,and the apparent film thickness decreased significantly from 1.26 cm to 1.08 cm,which was 14.29% thinner(p<0.05). LF-EMF radiation improved the reaction efficiency of enzymatic starch hydrolysis catalyzed by α-amylase,one of the reason should be the enhancement of mass transfer,but the possibility could not be excluded that the LF-EMF radiation might affect the enzyme's conformation and thus increase its activity.
To investigate the effect and mechanism of low frequency electromagnetic field(LF-EMF)on enzymatic reaction,in this paper,the hydrolysis of corn starch catalyzed by α-amylase was used as a model reaction,treatment on the reaction with 60 Hz LF-EMF was carried out. Based on the analyzing of the LF-EMF's influence on the reaction efficiency of enzymatic starch hydrolysis,and its influence on the substrate structure,the enzyme structure and the mass transfer in solution system,the mechanism of the LF-EMF's influence on the enzymatic reaction was discussed preliminarily. The results showed that LF-EMF radiation promoted the hydrolysis of starch catalyzed by α-amylase,14.39 mg/mL hydrolyzed sugar was produced in 10 h reaction,which was 15.90% higher than the control(12.41 mg/mL)(p<0.05);Neither remarkable change on the primary structure of α-amylase,nor irreversible change on the secondary structure/tertiary structure resulted from LF-EMF radiation was observed(p>0.05). The LF-EMF could reduce the optimum ionic strength(which was 0.0947 mol/L)of the α-amylase's reaction system,and weaken the inhibition of high ionic strength on the enzymatic reaction. The sodium chloride diffusion experiment indicated that LF-EMF could promote mass transfer in solution system,the enhancement factor of ion diffusion was 1.23,and the apparent film thickness decreased significantly from 1.26 cm to 1.08 cm,which was 14.29% thinner(p<0.05). LF-EMF radiation improved the reaction efficiency of enzymatic starch hydrolysis catalyzed by α-amylase,one of the reason should be the enhancement of mass transfer,but the possibility could not be excluded that the LF-EMF radiation might affect the enzyme's conformation and thus increase its activity.
引文
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[11]蔡杏华,任浩锋,蒙永双,等.低频静电场辐射对酿酒酵母生长代谢的影响[J].食品工业科技,2018(2):124-127.
[12]Xiang-Jun Q I,Gou J X,Han X J,et al.Study on measuring reducing sugar by DNS reagent[J].Journal of Cellulose Science & Technology,2004,12(3):17-445.
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[14]陈翠兰,张本山,陈福泉.淀粉结晶度计算的新方法[J].食品科学,2011,32(9):68-71.
[15]U.K.LaemmLi.Cleavage of structural proteins during the assembly of the head of bacteriophage T4[J].Nature,1970,227,680-685.
[16]Chen L,Liang J F.Peptide fibrils with altered stability,activity,and cell selectivity[J].Biomacromolecules,2013,14(7):2326-2331.
[17]Spotti M J,Martinez M J,Pilosof A M R,et al.Influence of Maillard conjugation on structural characteristics and rheological properties of whey protein/dextran systems[J].Food Hydrocolloids,2014,39(39):223-230.
[18]Levanda M,Fleurov V.A Wigner quasi-distribution function for charged particles in classical electromagnetic fields[J].Annals of Physics,2001,292(2):199-231.
[19]何魏魏,王萍,赵果,等.紫菜藻胆蛋白质的提取纯化与抗病毒研究[J].齐鲁渔业,2012(7):4-6.
[20]张太平.磷酸盐缓冲溶液(Na2HPO4-NaH2PO4)pH计算公式及其应用[J].高等继续教育学报,2001,14(2):25-26.
[21]张惠.超声波对传质过程强化的研究[D].杭州:浙江工业大学,2002.
[22]张亚楠,薛济来,朱骏,等.超声功率对水溶液中NaCl颗粒溶解速率的影响[J].中国科技论文,2014(12):1336-1339.
[23]喻佩,于淑娟,孙炜炜.高静压对酪蛋白-半乳糖美拉德反应体系的影响[J].食品工业科技,2012,33(4):188-191.
[24]张春玲,赵长新,董亮,张帆,邵国壮.大麦发芽过程中金属离子对大麦酶系中淀粉酶活力影响研究[J].食品工业科技,2005(8):75-77,81.
[25]Mesbah N M,Wiegel J.Halophilic alkali-and thermostable amylase from a novel polyextremophilic Amphibacillus sp.NM-Ra2[J].International Journal of Biological Macromolecules.September 2014,70:222-229.
[26]Deshpande S S,Cheryan M.Effects of phytic acid,divalent cations,and their interactions on α-amylase activity[J].Journal of Food Science,2010,49(2):516-519.
[27]聂国兴,明红,胡建业,等.无机离子对淀粉酶活性的影响[J].水生态学杂志,2004,24(4):26-29.
[28]方华,黄俊,陈瞾,等.缓冲体系和金属离子对漆酶催化活性的影响[J].武汉理工大学学报,2009(5):27-30.
[29]Wu H,Niu L,Xu Y.In-situ synthesis of WC particles via electromagnetic field[J].Journal of the Chinese Ceramic Society,2010,38(8):1591-1595.
[2]林亲录,周丽君,符琼.淀粉转化生产葡萄糖工艺研究进展[J].食品工业科技,2011(4):412-414.
[3]贺福元,邓凯文,刘文龙,等.葡萄糖应用生产现状及碎(大)米酶法制成注射用葡萄糖的巨大优势[J].湖南中医药大学学报,2010,30(12):68-71.
[4]刘力倩.我国玉米淀粉糖生产历程综述[J].粮食流通技术,2017(1):72-74.
[5]刘洪,张小云,谭丽,等.微波-双酶耦合催化淀粉水解[J].食品与发酵工业,2010(4):101-103.
[6]石文奇,易长海,甘厚磊,等.超声波对液体α-淀粉酶酶学活性的影响[J].天津工业大学学报,2012,31(2):43-46.
[7]徐茜,曹毅.极低频电磁场与儿童白血病关系的研究进展[J].中国辐射卫生,2009,18(2):243-245.
[8]薛丽萍,孟亚萍,赵娟娟,等.极低频交变磁场对过氧化氢酶活性的影响[J].食品与生物技术学报,2014,33(3):268-274.
[9]张秋霞,尹春玲,范秋领,朱元保.磁场对脂肪酶催化活性的影响[J].平顶山学院学报,2005(5):59-61.
[10]肖祖峰,陈明东,韩光泽.电磁场作用下的强化传质研究进展[J].化工进展,2008,27(12):1911-1916.
[11]蔡杏华,任浩锋,蒙永双,等.低频静电场辐射对酿酒酵母生长代谢的影响[J].食品工业科技,2018(2):124-127.
[12]Xiang-Jun Q I,Gou J X,Han X J,et al.Study on measuring reducing sugar by DNS reagent[J].Journal of Cellulose Science & Technology,2004,12(3):17-445.
[13]赵蓉,李多伟,任涛,等.DNS比色法测定白芸豆中α-淀粉酶抑制剂活性的方法研究[J].中成药,2013,35(3):573-576.
[14]陈翠兰,张本山,陈福泉.淀粉结晶度计算的新方法[J].食品科学,2011,32(9):68-71.
[15]U.K.LaemmLi.Cleavage of structural proteins during the assembly of the head of bacteriophage T4[J].Nature,1970,227,680-685.
[16]Chen L,Liang J F.Peptide fibrils with altered stability,activity,and cell selectivity[J].Biomacromolecules,2013,14(7):2326-2331.
[17]Spotti M J,Martinez M J,Pilosof A M R,et al.Influence of Maillard conjugation on structural characteristics and rheological properties of whey protein/dextran systems[J].Food Hydrocolloids,2014,39(39):223-230.
[18]Levanda M,Fleurov V.A Wigner quasi-distribution function for charged particles in classical electromagnetic fields[J].Annals of Physics,2001,292(2):199-231.
[19]何魏魏,王萍,赵果,等.紫菜藻胆蛋白质的提取纯化与抗病毒研究[J].齐鲁渔业,2012(7):4-6.
[20]张太平.磷酸盐缓冲溶液(Na2HPO4-NaH2PO4)pH计算公式及其应用[J].高等继续教育学报,2001,14(2):25-26.
[21]张惠.超声波对传质过程强化的研究[D].杭州:浙江工业大学,2002.
[22]张亚楠,薛济来,朱骏,等.超声功率对水溶液中NaCl颗粒溶解速率的影响[J].中国科技论文,2014(12):1336-1339.
[23]喻佩,于淑娟,孙炜炜.高静压对酪蛋白-半乳糖美拉德反应体系的影响[J].食品工业科技,2012,33(4):188-191.
[24]张春玲,赵长新,董亮,张帆,邵国壮.大麦发芽过程中金属离子对大麦酶系中淀粉酶活力影响研究[J].食品工业科技,2005(8):75-77,81.
[25]Mesbah N M,Wiegel J.Halophilic alkali-and thermostable amylase from a novel polyextremophilic Amphibacillus sp.NM-Ra2[J].International Journal of Biological Macromolecules.September 2014,70:222-229.
[26]Deshpande S S,Cheryan M.Effects of phytic acid,divalent cations,and their interactions on α-amylase activity[J].Journal of Food Science,2010,49(2):516-519.
[27]聂国兴,明红,胡建业,等.无机离子对淀粉酶活性的影响[J].水生态学杂志,2004,24(4):26-29.
[28]方华,黄俊,陈瞾,等.缓冲体系和金属离子对漆酶催化活性的影响[J].武汉理工大学学报,2009(5):27-30.
[29]Wu H,Niu L,Xu Y.In-situ synthesis of WC particles via electromagnetic field[J].Journal of the Chinese Ceramic Society,2010,38(8):1591-1595.
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