具有DPPH自由基抑制活性的蚕蛹蛋白酶解液脱色工艺优化
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摘要
利用木瓜蛋白酶水解的蚕蛹蛋白酶解液作为研究原料,根据响应面试验设计方法,以脱色率和1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除保持率作为响应面指标,得最佳脱色工艺条件为:活性白土、碱性钙基膨润土和膨润土按照质量比1∶1∶1混合为脱色剂、脱色剂用量1.9%、酶解液质量浓度9.61 mg/m L、p H 7.2、水浴温度60℃、振荡转速150 r/min、脱色时间1.0 h。在此条件下实验,得出DPPH自由基清除保持率84.5%、脱色率94.5%,与预测值相比误差值均小于5%,说明采用响应面试验设计方法对蚕蛹蛋白酶解液脱色体系进行优化具有较好的可靠性。最后再对脱色前后酶解液中各氨基酸含量进行比较,发现经脱色处理后酶解液中对DPPH自由基抑制有显著作用的疏水性氨基酸和芳香族氨基酸含量减少。
Papain hydrolyzate of silkworm pupae protein with DPPH radical scavenging activity was decolorized by the combined use of activated clay, alkaline Ca-bentonite and bentonite. Based on decolorization efficiency and retention rate of radical scavenging activity, the decolorization conditions were optimized by using single-factor design and central composite design(CCD) coupled with response surface methodology(RSM). When the hydrolyzate with a concentration of 9.61 mg/m L and an initial p H of 7.2 was added with activated clay:alkaline Ca-bentonite:bentonite(1:1:1) in a proportion of 1.9% and incubated for 1.0 h in a thermostatically controlled water at 60 ℃ with shaking at a speed of 150 r/min, it retained 84.5%of its original DPPH radical scavenging activity and the decolorization rate was 94.5%, both close to the predicted values with a relative error less than 5%, suggesting the reliability of the developed response surface model. Comparing the amino acid composition of the decolorized hydrolyzate with that of the untreated one, it was found that the contents of hydrophobic and aromatic amino acids which are significantly effective in inhibiting DPPH free radical were reduced after the decolorization treatment.
Papain hydrolyzate of silkworm pupae protein with DPPH radical scavenging activity was decolorized by the combined use of activated clay, alkaline Ca-bentonite and bentonite. Based on decolorization efficiency and retention rate of radical scavenging activity, the decolorization conditions were optimized by using single-factor design and central composite design(CCD) coupled with response surface methodology(RSM). When the hydrolyzate with a concentration of 9.61 mg/m L and an initial p H of 7.2 was added with activated clay:alkaline Ca-bentonite:bentonite(1:1:1) in a proportion of 1.9% and incubated for 1.0 h in a thermostatically controlled water at 60 ℃ with shaking at a speed of 150 r/min, it retained 84.5%of its original DPPH radical scavenging activity and the decolorization rate was 94.5%, both close to the predicted values with a relative error less than 5%, suggesting the reliability of the developed response surface model. Comparing the amino acid composition of the decolorized hydrolyzate with that of the untreated one, it was found that the contents of hydrophobic and aromatic amino acids which are significantly effective in inhibiting DPPH free radical were reduced after the decolorization treatment.
引文
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[2]胡建平,刘可桃.蚕蛹蛋白的提取工艺及综合利用研究进展[J].安徽农学通报,2011,17(11):185-186.
[3]李国荣,张华峰,李建科,等.蚕蛋白质资源的研究现状及其在食品工业中的应用[J].食品工业科技,2012,33(5):396-401.
[4]闵建华,李建科,陈婷.蚕蛹多肽的制备工艺及其体外抗氧化活性[J].食品科学,2009,30(14):123-126.
[5]赵钟兴,廖丹葵,孙建华,等.蚕蛹蛋白酶解产物体外抗氧化和降血压活性筛选及响应面工艺优化[J].食品科学,2011,32(23):186-191.
[6]吴晓霞,李建科,张研宇.蚕蛹油超声波辅助萃取及其抗氧化稳定性[J].中国农业科学,2010,43(8):1677-1687.
[7]章绍兵,甘晓露,龚琴.花生蛋白酶解液的活性炭脱色工艺研究[J].河南工业大学学报:自然科学版,2011,32(2):10-14.
[8]张立峰,刘庆富,宁海风.活性炭对大豆低聚糖脱色效果研究[J].粮食与油脂,2013,26(1):51-53.
[9]马永香,华欲飞,孔祥珍,等.离子交换树脂对大豆糖蜜上清液脱盐脱色工艺的研究[J].中国油脂,2013,38(1):70-75.
[10]LIU Jun,LUO Jiangguang,SUN Yi,et al.A simple method for the simultaneous decoloration and deproteinization of crude levan extract from Paenibacillus polymyxa EJS-3 by macroporous resin[J].Bioresource Technology,2010,101(15):6077-6083.
[11]赵华,王虹,任晶,等.响应面法提取玉米醇溶蛋白的工艺优化[J].食品研究与开发,2012,33(5):38-44.
[12]郑仕远,谢志刚,曹优明,等.蚕蛹的溶剂法一步脱脂、脱色、除臭工艺研究[J].食品科技,2003,24(8):49-50.
[13]韦藤幼,童张法,吴璇.碱性钙基膨润土及其制备方法:中国,ZL200510101332.X[P].2009-07-08.
[14]赵娟,韦藤幼,潘远凤,等.碱性白土的制备及其脂肪酸脱除机理研究[J].广西大学学报,2011,36(3):379-384.
[15]ZHANG Hanbing,TONG Zhangfa,WEI Tengyou,et al.Removal characteristics of Zn(Ⅱ)from aqueous solution by alkaline Cabentonite[J].Desalination,2011,276:103-108.
[16]张寒冰,胡雪玲,韦藤幼,等.碱性钙基膨润土的制备及其应用研究进展[J].化工进展,2012,31(7):1395-1401.
[17]涂宗财,王艳敏,迟海霞,等.响应面法优化豆豉蛋白酶解工艺[J].食品与发酵工业,2009,35(6):116-120.
[18]张海祥,魏兆军,周乐春,等.响应曲面法优化动物蛋白酶水解脱脂蚕蛹蛋白的工艺[J].食品科学,2009,30(24):188-192.
[19]刘石生,林志锋,周国英,等.响应面法优化罗非鱼肉内源酶酶解工艺[J].现代食品科技,2010,26(1):85-88.
[20]章绍兵,甘晓露,龚琴.琴花生蛋白酶解液的活性炭脱色工艺研究[J].河南工业大学学报:自然科学版,2011,32(2):10-14.
[21]吕汶骏,赵钟兴,廖丹葵,等.蚕蛹蛋白及其水解产物中氨基酸组成分析[J].食品科学,2012,33(12):228-232.
[22]秦伟佳,赵钟兴,吕汶骏,等.蚕蛹蛋白精制及其氨基酸成分分析[J].食品工业科技,2013,34(13):197-201.
[23]郑为完,徐慧诠,周小娟,等.可溶性米渣水解蛋白粉的脱色工艺[J].南昌大学学报:工科版,2009,31(1):14-17.
[24]赵欢欢,刘玉兰,张小涛,等.吸附法脱除芝麻油中苯并芘及脱色效果研究[J].粮油食品科技,2014,21(4):23-27.
[25]沈芸,翁新楚,宋红生,等.凹凸棒土对蚕蛹油的脱色[J].上海大学学报:自然科学版,2011,17(5):687-690.
[26]霍英霞.豆油碱性白土脱酸工艺及动力学研究[D].南宁:广西大学,2013.
[27]REN Jiaoyan,ZHAO Mouming,SHI J,et al.Purification and identification of antioxidant peptides from grass carp muscle hydrolysates by consecutive chromatography and electrospray ionizationmass spectrometry[J].Food Chemistry,2008,108(2):727-736.
[28]KIM S K,KIM Y T,BYUM H G,et al.Isolation and characterization of antioxidative peptides from gelatin hydrolysate of Alaska pollack skin[J].Journal of Agricultural and Food Chemistry,2001,49(4):1984-1989.
[29]LI Yaowang,LI Bo,HE Jiguo,et al.Structure-activity relationship study of antioxidative peptides by QSAR modeling:the amino acid next to C-terminus affects the activity[J].Journal of Peptide Science,2011,17(6):454-462.
[30]JIANG Haiping,TONG Tianzhe,SUN Jianghua,et al.Purification and characterization of antioxidative peptides from round scad(Decapterus maruadsi)muscle protein hydrolysate[J].Food Chemistry,2014,154(5):158-163.