Effect of pH on the conformational structure of cytochrome c and subsequent enzymatic cross-linking catalyzed by laccase

被引：0

作者：

Li D.-X. ^{[1
]}

Qi Z.-Y. ^{[1
]}

Liu J.-Y. ^{[1
]}

Zhou J.-Q. ^{[1
]}

机构：

[1] College of Pharmaceutical Sciences, Soochow University, Suzhou

来源：

Electronic Journal of Biotechnology | 2022年 / 60卷

基金：

中国国家自然科学基金;

关键词：

Conformational structure; Cytochrome c; High-performance size exclusion chromatography; Isothermal titration calorimetry; Laccase; Laser particle size analysis; pH value; Protein cross-linking; Substrate requirement;

D O I：

10.1016/j.ejbt.2022.07.002

中图分类号：

学科分类号：

摘要：

Background: The aim of the present study was to investigate the effect of substrate conformational structure changes on the laccase-induced protein cross-linking. The effects of laccase amount, pH, and ferulic acid (FA) on the enzymatic cross-linking of substrate, Cyt C, were determined by sodium dodecyl sulfate–polyacrylamide gel electrophoresis. High-performance size exclusion chromatography, laser particle size analysis and isothermal titration calorimetry (ITC) were also applied to investigate the cross-linking product and enthalpy changes. Structural changes of Cyt C at different pH values were analyzed by ultraviolet–visible (UV–vis), fluorescence, and circular dichroism (CD) measurements. Results: Complete cross-linking, partial cross-linking, minute cross-linking, and no cross-linking occurred at pH 2.0, 4.0, 6.0, and 8.0, respectively. ITC analysis demonstrated that the enzymatic cross-linking of Cyt C was an endothermic process. The UV–vis, fluorescence, and CD measurements exhibited that the tertiary structure of Cyt C was disrupted, and part of the α-helical polypeptide region unfolded at pH 2.0. The structural flexibilities decreased, and the tertiary structure of Cyt C became increasingly compact with the increase in pH values from 4.0 to 8.0. The gradual changes in the structure of Cyt C at different pH values were in accordance with the cross-linking results of Cyt C catalyzed by laccase. Conclusions: The results demonstrated that minute structure changes of substrate had a remarkable effect on the laccase-induced cross-linking. The findings promote the understanding of the substrate requirement of laccase in protein cross-linking and are instructive for the modulation of laccase-induced protein cross-linking. How to cite: Li D-X, Qi Z-Y, Liu J-Y, et al. Effect of pH on the conformational structure of cytochrome c and subsequent enzymatic cross-linking catalyzed by laccase. Electron J Biotechnol 2022;60. https://doi.org/10.1016/j.ejbt.2022.07.002. © 2022

引用

页码：1 / 10

页数：9

共 40 条

[1]

Isaschar-Ovdat S., Davidovich-Pinhas M., Fishman A., Modulating the gel properties of soy glycinin by crosslinking with tyrosinase, Food Res Int, 87, pp. 42-49, (2016)

[2]

Yuan X.Y., Li X.Y., Zhang X.L., Et al., Effect of ultrasound on structure and functional properties of laccase-catalyzed α-lactalbumin, J Food Eng, 223, pp. 116-123, (2018)

[3]

Teixeira L.S.M., Feijen J., van Blitterswijk C.A., Et al., Enzyme-catalyzed crosslinkable hydrogels: emerging strategies for tissue engineering, Biomater, 33, pp. 1281-1290, (2012)

[4]

Xu D.Y., Yang Y., Yang Z., Activity and stability of cross-linked tyrosinase aggregates in aqueous and nonaqueous media, J Biotechnol, 152, pp. 30-36, (2011)

[5]

Hu H., Zhu X.R., Hu T., Et al., Effect of ultrasound pre-treatment on formation of transglutaminase-catalysed soy protein hydrogel as a riboflavin vehicle for functional foods, J Funct Foods, 19, pp. 182-193, (2015)

[6]

Elzoghby A.O., El-Fotoh W.S.A., Elgindy N.A., Casein-based formulations as promising controlled release drug delivery systems, J Control Release, 153, pp. 206-216, (2011)

[7]

Fan L.H., Wu H., Zhou X.Y., Et al., Transglutaminase-catalyzed grafting collagen on chitosan and its characterization, Carbohydr Polym, 105, pp. 253-259, (2014)

[8]

Zhou J.Q., He T., Wang J.W., The microbial transglutaminase immobilization on carboxylated poly(N-isopropylacrylamide) for thermo-responsivity, Enzyme Microb Technol, 87-88, pp. 44-51, (2016)

[9]

Faccio G., Kruus K., Saloheimo M., Et al., Bacterial tyrosinases and their applications, Process Biochem, 47, pp. 1749-1760, (2012)

[10]

Garcia-Jimenez A., Teruel-Puche J.A., Garcia-Ruiz P.A., Et al., Action of tyrosinase on caffeic acid and its n-nonyl ester. Catalysis and suicide inactivation, Int J Biol Macromol, 107, pp. 2650-2659, (2018)

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