纤维素酶与表面活性剂的相互作用及其在洗涤剂中的应用

doi:10.11949/j.issn.0438-1157.20151989

摘要/Abstract

摘要：

研究了表面活性剂对纤维素酶活力的影响，并通过动态光散射（DLS）、荧光光谱及傅里叶变换衰减全反射红外光谱（ATR-FTIR）探讨了表面活性剂和纤维素酶的相互作用机理。DLS实验结果显示，阴离子表面活性剂吸附到了纤维素酶表面，使得阴离子表面活性剂-纤维素酶体系中的zeta电位分布在较低的负电荷区域；而非离子表面活性剂-纤维素酶与蒸馏水-纤维素酶体系中的zeta电位的分布没有明显的区别。荧光和ATR-FTIR实验说明了阴离子表面活性剂对纤维素酶的二级结构和侧链的微环境都有很大的影响。此外，对比了添加与未添加纤维素酶的洗衣液对国标污布JB-03的去污力，通过光学显微镜及扫描电镜（SEM）观察国标污布洗涤前后织物表面，发现将纤维素酶添加到洗衣液能明显提高洗衣液的去污能力。

关键词: 纤维素酶, 表面活性剂, 动态光散射, 荧光光谱, ATR-FTIR, 去污力

Abstract:

Effects of surfactants on cellulase activity were studied. The interaction mechanism between surfactants and cellulase was discussed by dynamic light scattering (DLS), fluorescence spectrum and ATR-FTIR. The DLS results showed that anionic surfactants were adsorbed onto cellulase surface, which made cellulase zeta potential distribute in the lower negative charge region. There was no obvious difference of cellulase zeta potential between in nonionic surfactants and deionized water. As shown in spectroscopy, the anionic surfactants had stronger influence on the main chain and side chain of cellulase than nonionic surfactant AEOs. The washing performance data of liquid detergent with and without cellulase indicated that adding cellulase into liquid detergent can enhance detergency of liquid detergent.

Key words: cellulase, surfactant, DLS, fluorescence spectrum, ATR-FTIR, detersive power

中图分类号:

TQ028.8

于跃, 张剑. 纤维素酶与表面活性剂的相互作用及其在洗涤剂中的应用[J]. 化工学报, 2016, 67(7): 3023-3031.

YU Yue, ZHANG Jian. Interaction of cellulase with surfactants and their application in detergent[J]. CIESC Journal, 2016, 67(7): 3023-3031.

参考文献 34

[1]	PEREZ J, MUNOZ-DORADO J, DE LA RUBIA T, et al. Biodegradation and biological treatments of cellulose, hemicelluloses and lignin an overview [J]. Int. Microbiol., 2002, 5: 53-63. DOI: 10.1007/s10123-002-0062-3.
[2]	BHAT M K, BHAT S. Cellulose degrading enzymes and their potential industrial applications [J]. Biotechnol. Adv., 1997, 15: 583-620. DOI: 10.1016/S0734-9750(97)00006-2.
[3]	项瑾, 梁毅, 陈楠. 等温滴定量热法和荧光滴定法研究十二烷基硫酸钠与纤维素酶的结合[J]. 化学学报, 2003, 61(12): 1949-1954. XIANG J, LIANG Y, CHEN N. Studies on the binding of sodium dodecyl sulfate to cellulase by isothermal titration calorimetry and fluorescence titration [J].Acta Chimica Sinica, 2003, 61(12): 1949-1954.
[4]	MANDELS M. Applications of cellulases [J]. Biochem. Soc. Trans., 1985, 13: 414-416.
[5]	BEGUIN P, AUBERT J P. The biological degradation of cellulose [J]. FEMS Microbiol., 1994, 13: 25-58. DOI: 10.1016/0168- 6445(94)90099-X.
[6]	LEE I, EVANS B R, WOODWARD J. The mechanism of cellulase action on cotton fibers: evidence from atomic force microscopy [J]. Ultramicroscopy, 2000, 82: 213-221. DOI: 10.1016/S0304- 3991(99)00158-8.
[7]	LIANG Y S, YUAN X Z, ZENG G M, et al. Effects of surfactants on enzyme-containing reversed micellar system [J]. Sci. China Chem., 2011, 54: 715-723. DOI: 10.1007/s11426-011-4266-2.
[8]	于跃, 张剑. 纤维素酶降解纤维素机理的研究进展[J]. 化学通报, 2016, 79(2): 118-122. YU Y, ZHANG J. Research progress in cellulose degradation by cellulase [J]. Chemistry Online, 2016, 79(2): 118-122.
[9]	和田恭尚. 酶在洗涤剂中的应用现状及展望[J]. 日用化学工业, 2005, 35(1): 30-35. WADA Y. Status and prospect of application of enzyme in detergent products [J]. China Surfactant Detergent & Cosmetics, 2005, 35(1):30-35.
[10]	HOSHINO E, CHIWAKI M, SUZUKI A, et al. Improvement of cotton cloth soil removal by inclusion of alkaline cellulase from Bacillus sp. KSM-635 in detergents [J]. Journal of Surfactant and Detergents, 2000, 3: 317-326.
[11]	VASCONCELOS A, CAVACO-PAULO A. Enzymatic removal of cellulose from cotton/polyester fabric blends [J]. Cellulose, 2006, 13: 611-618. DOI: 10.1007/s10570-06-9063-2.
[12]	MIZUTANI C, SETHUMASHAVAN K, HOWLEY P, et al. Effect of a nonionic surfactant on Trichoderma cellulase treatments of regenerated cellulose and cotton yarns [J]. Cellulose, 2002, 9: 83-89.
[13]	姚兰, 赵建, 谢益民, 等.木质素结构以及表面活性剂对木质素吸附纤维素酶的影响[J]. 化工学报, 2012, 63(8): 2612-2616. YAO L, ZHAO J, XIE Y M, et al. Effect of lignin structure and surfactant on cellulase adsorption by lignin [J]. CIESC Journal, 2012, 63(8): 2612-2616.
[14]	XIA J, CHEN X, NNANNA I A. Activity and stability of Penicillium cyclopium lipase in surfactant and detergent solutions [J]. J. Am. Chem. Soc., 1996: 115-20. DOI: 10.1007/BF02523457.
[15]	沈兵兵.复合酶在液体洗涤剂中的应用[D]. 上海: 华南理工大学, 2012: 32-33. SHEN B B. Application of composite enzymes in liquid laundry detergent [D]. Shanghai: South China University of Technology, 2012: 32-33.
[16]	COX M F, 吴姣莲. 烷基链长和EO含量对AES性能的影响[J]. 日用化学品科学, 1990, (5): 1-9. COX M F, WU J L. Effect of alkyl chain length and EO chain on AES [J]. Detergent & Cosmetics, 1990, (5): 1-9.
[17]	史兴旺. 新颖表面活性剂对牛血清蛋白(BSA)结构的影响研究[D]. 济南: 山东大学, 2008: 49-51. SHI X W. Studies on the effects of novel surfactants on the structures of bovine serum albumin(BSA) [D]. Jinan: Shandong University, 2008: 49-51.
[18]	叶进富. 蛋白质Zeta电位与离子交换层分析的相关性研究[D]. 杭州: 浙江大学, 2006: 33-35. YE J F. Zeta potential of protein and its correlation to separation behavior of ion exchange chromatography [D]. Hangzhou: Zhejiang University, 2006: 33-35.
[19]	刘微, 李萌, 任皓威, 等. 荧光、紫外和红外光谱分析人乳和牛乳β-酪蛋白的功能和构象差异[J]. 光谱学与光谱分析, 2014, 34(12): 3283-3287. DOI: 10.3964/j.issn.100-0593(2014)12-3281-07. LIU W, LI M, RENG H W, et al. Comparison of the function and conformation of human β-casein and bovine β-casein by spectroscopic study [J]. Spectroscopy and Spectral Analysis, 2014, 34(12): 3283-3287. DOI: 10.3964/j.issn.100-0593(2014)12-3281-07.
[20]	刘媛, 谢孟峡, 康娟. 三七总皂甙对牛血清白蛋白溶液构象的影响[J]. 化学学报, 2003, 61(8): 1305-1310. LIU Y, XIE M X, KANG J. Influence of total saponins of Panax notoginseng on the conformation of BSA[J]. Acta Chimica Sinica, 2003, 61(8): 1305-1310.
[21]	马萍, 迟燕华, 庄稼, 等. 骨螺紫及其铜配合物与人血清白蛋白相互作用的光谱学研究[J]. 高等学校化学学报, 2009, (8): 1509-1515. MA P, CHI Y H, ZHUANG J, et al. Spectroscopic studies on the interaction of murexide and murexide-copper complex with human serum albumin [J].Chemical Journal of Chinese Universities, 2009, (8): 1509-1515.
[22]	马林, 魏志强, 黄爱民, 等. 丙三醇对水溶液中血红蛋白构象的影响-荧光猝灭和动态光散射研究[J]. 化学学报, 2009, 67(14): 1566-1572. MA L, WEI Z Q, HUANG A M, et al. Influence of glycerol on the conformation of hemoglobin in aqueous solutions: a study of fluorescence quenching and dynamic light scattering [J]. Acta Chimica Sinica, 2009, 67(14): 1566-1572.
[23]	魏晓芳, 西明, 刘会洲. pH诱导牛血清白蛋白芳香氨基酸残基微环境变化的光谱分析[J]. 光谱学与光谱分析, 2000, 20(4): 556-559. DOI: 10.3321/j.issn:1000-0593.2000.04.028. WEI X F, XI M, LIU H Z.Spectral study of the microenviroment change of aromatic amino-acid residues in BSA induced by pH [J]. Spectroscopy and Spectral Analysis, 2000, 20(4): 556-559. DOI: 10.3321/j.issn:1000-0593.2000.04.028.
[24]	NIELSEN A D, ARLETH L, WESTH P. Analysis of protein- surfactant interactions - a titration calorimetric and fluorescence spectroscopic investigation of interactions between Humicola insolens cutinase and an anionic surfactant [J]. Biochimica et Biophysica Acta, 2005, (1752): 124-132. DOI: 10.1016/j.bbapap.2005.08.001.
[25]	BROCKHINKE A, PLESSOW R, DITTRICH P, et al. Analysis of the local conformation of proteins with two-dimensional fluorescence techniques [J]. Appl. Phys., 2000, (71): 755-763. DOI: 10.1007/ s00340000384.
[26]	LAKOWICZ J R. Principles of Fluorescence Spectroscopy [M]. 3rd ed. Springer, 2006: 529-575.
[27]	VONGSVIVUT J, HERAUD P, ZHANG W, et al. Rapid determination of protein contents in microencapsulated fish oil supplements by ATR-FTIR spectroscopy and partial least square regression (PLSR) analysis [J]. Food and Bioprocess Technology, 2014, 7: 265-277. DOI: 10.1007/s11947-013-1122-8.
[28]	袁久刚, 文艺. 表面活性剂对纤维素酶水洗性能的影响[J]. 纺织学报, 2014, 35(5): 78-82. DOI: 10.13475/j.fzxb.201405007805. YUAN J G, WEN Y. Effect of surfactant on cellulase bio-washing [J]. Journal of Textile Research, 2014, 35(5): 78-82. DOI: 10.13475/j.fzxb. 201405007805.
[29]	余多慰, 袁生. 绿色木霉纤维素酶分子内氢键特征的研究[J]. 光谱学与光谱分析, 2005, 25(4): 544-547. YU D W, YUAN S. Hydrogen-bond state analysis of cellobiohydrolase Ⅱ molecule from Trichoderma viride [J]. Spectroscopy and Spectral Analysis, 2005, 25(4): 544-547.
[30]	ZHANG J, ZHANG J. Study on the interaction of alkaline protease with main surfactants in detergent [J]. Colloid and Polymer Science, 2015, 9: 1-9. DOI: 10.1007/s00396-015-3777-3.
[31]	GUILLEN M D, CABO N. Characterization of edible oils and lard by Fourier transform infrared spectroscopy. Relationships between composition and frequency of concrete bands in the fingerprint region [J]. J. Am. Chem. Soc., 1997, 74: 1281-1286. DOI: 10.1007/ s11746-997-0058-4.
[32]	董荣, 李江华, 陈银生. 液体洗涤剂组分对青霉脂肪酶活性的影响[J]. 日用化学品科学, 2000, 23(S1): 100-102. DONG R, LI J H, CHEN Y S. Effects of liquid detergent components on activity of penicillium lipase [J]. Detergent & Cosmetics, 2000, 23(S1): 100-102.
[33]	IVANOV I B, KRALCHEVSKY P A. Stability of emulsions under equilibrium and dynamic conditions [J]. Colloids and Surfaces A, 1997: 155.
[34]	梁运姗, 袁兴中, 曾光明, 等. 表面活性剂在逆胶束酶反应系统中的作用机制[J]. 中国科学: 化学, 2011, 41(5): 763-775. LIANG Y S, YUAN X Z, ZENG G M, et al. Effects of the surfactants on enzyme-containing reversed micellar system [J]. Scientia Sinica Chimica, 2011, 41(5): 763-775.