A Novel Gelatinase from Marine Flocculibacter collagenilyticus SM1988: Characterization and Potential Application in Collagen Oligopeptide-Rich Hydrolysate Preparation

• Published in: Marine drugs Vol. 20; no. 1; p. 48
• Main Authors: Li, Jian, Cheng, Jun-Hui, Teng, Zhao-Jie, Zhang, Xia, Chen, Xiu-Lan, Sun, Mei-Ling, Wang, Jing-Ping, Zhang, Yu-Zhong, Ding, Jun-Mei, Tian, Xin-Min, Zhang, Xi-Ying
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 03.01.2022; MDPI
• Subjects: Amino acids; Animals; Aquatic Organisms; Bones; bovine bone collagen; Cattle; Chromatography; Collagen; Collagen - chemistry; E coli; Enzymes; Enzymolysis; Gelatin; Gelatinase; Gelatinases - chemistry; Gelatinases - pharmacology; Genomes; hydrolysate; Hydrolysates; Hydrolysis; Molecular weight; Oligopeptides; Oligopeptides - chemistry; Peptidase; Peptidases; Peptides; Proteins; Proteobacteria; Recombinants; Skin; Structure-Activity Relationship; Substrates; the MEROPS S8 family; hydrolysate; bovine bone collagen; the MEROPS S8 family; oligopeptides; peptidase
• ISSN: 1660-3397; 1660-3397
• DOI: 10.3390/md20010048

Although the S8 family in the MEROPS database contains many peptidases, only a few S8 peptidases have been applied in the preparation of bioactive oligopeptides. Bovine bone collagen is a good source for preparing collagen oligopeptides, but has been so far rarely applied in collagen peptide preparation. Here, we characterized a novel S8 gelatinase, Aa2_1884, from marine bacterium SM1988 , and evaluated its potential application in the preparation of collagen oligopeptides from bovine bone collagen. Aa2_1884 is a multimodular S8 peptidase with a distinct domain architecture from other reported peptidases. The recombinant Aa2_1884 over-expressed in showed high activity toward gelatin and denatured collagens, but no activity toward natural collagens, indicating that Aa2_1884 is a gelatinase. To evaluate the potential of Aa2_1884 in the preparation of collagen oligopeptides from bovine bone collagen, three enzymatic hydrolysis parameters, hydrolysis temperature, hydrolysis time and enzyme-substrate ratio (E/S), were optimized by single factor experiments, and the optimal hydrolysis conditions were determined to be reaction at 60 ℃ for 3 h with an E/S of 400 U/g. Under these conditions, the hydrolysis efficiency of bovine bone collagen by Aa2_1884 reached 95.3%. The resultant hydrolysate contained 97.8% peptides, in which peptides with a molecular weight lower than 1000 Da and 500 Da accounted for 55.1% and 39.5%, respectively, indicating that the hydrolysate was rich in oligopeptides. These results indicate that Aa2_1884 likely has a promising potential application in the preparation of collagen oligopeptide-rich hydrolysate from bovine bone collagen, which may provide a feasible way for the high-value utilization of bovine bone collagen.