title: Effect of ultrasonic pretreatment on kinetics of gelatin hydrolysis by collagenase and its mechanism.
abstract: Gelatin is a mixture of soluble proteins prepared by partial hydrolysis of native collagen. Gelatin can be enzymatically hydrolyzed to produce bioactive hydrolysates. However, the preparation of gelatin peptide with expected activity is usually a time-consuming process. The production efficiency of gelatin hydrolysates needs to be improved. In present work, effect of ultrasonic pretreatment on kinetic parameters of gelatin hydrolysis by collagenase was investigated based on an established kinetic model. With ultrasonic pretreatment, reaction rate constant and enzyme inactivation constant were increased by 27.5% and 27.8%, respectively. Meanwhile, hydrolysis activation energy and enzyme inactivation energy were reduced by 36.3% and 43.0%, respectively. In order to explore its possible mechanism, influence of sonication on structural properties of gelatin was determined using atomic force microscopy, particle size analyzer, fluorescence spectroscopy, protein solubility test and Fourier transform infrared spectroscopy. Moreover, hydrogen peroxide was used as a positive control for potential sonochemical effect. It was found that reduction of gelatin particle size was mainly caused by physical effect of ultrasound. Increased solubility and variation in β-sheet and random coil elements of gelatin were due to sonochemical effect. Both physical and chemical effects of sonication contributed to the change in α-helix and β-turn structures. The current results suggest that ultrasound can be potentially applied to stimulate the production efficiency of gelatin peptides, mainly due to its effects on modification of protein structures.
authors.affiliation: The Key Laboratory of Leather Chemistry and Engineering of Ministry of Education, Sichuan University, Chengdu 610065, PR China; National Engineering Laboratory of Clean Technology for Leather Manufacture, Sichuan University, Chengdu 610065, PR China.
authors.forename: Zhi-Long
authors.initials: ZL
authors.lastname: Yu
authors.affiliation: The Key Laboratory of Leather Chemistry and Engineering of Ministry of Education, Sichuan University, Chengdu 610065, PR China.
authors.forename: Wei-Cai
authors.initials: WC
authors.lastname: Zeng
authors.affiliation: The Key Laboratory of Leather Chemistry and Engineering of Ministry of Education, Sichuan University, Chengdu 610065, PR China.
authors.forename: Wen-Hua
authors.initials: WH
authors.lastname: Zhang
authors.affiliation: The Key Laboratory of Leather Chemistry and Engineering of Ministry of Education, Sichuan University, Chengdu 610065, PR China.
authors.forename: Xue-Pin
authors.initials: XP
authors.lastname: Liao
authors.affiliation: The Key Laboratory of Leather Chemistry and Engineering of Ministry of Education, Sichuan University, Chengdu 610065, PR China; National Engineering Laboratory of Clean Technology for Leather Manufacture, Sichuan University, Chengdu 610065, PR China. Electronic address: shibi@scu.edu.cn.
authors.forename: Bi
authors.initials: B
authors.lastname: Shi
mesh_terms: D000818:Animals; D002417:Cattle; D017364:Collagenases / Q000378:metabolism*; D005780:Gelatin / Q000737:chemistry*; D006868:Hydrolysis; D007700:Kinetics; D010316:Particle Size; D013010:Sonication
chemical_list: D005780:Gelatin; D017364:Collagenases
keywords: Gelatin; Hydrolysis; Kinetics; Mechanism; Ultrasonic pretreatment