Potential of Thermolysin-like Protease A69 in Preparation of Bovine Collagen Peptides with Moisture-Retention Ability and Antioxidative Activity

Abstract

Bovine bone is rich in collagen and is a good material for collagen peptide preparation. Although thermolysin-like proteases (TLPs) have been applied in different fields, the potential of TLPs in preparing bioactive collagen peptides has rarely been evaluated. Here, we characterized a thermophilic TLP, A69, from a hydrothermal bacterium Anoxybacillus caldiproteolyticus 1A02591, and evaluated its potential in preparing bioactive collagen peptides. A69 showed the highest activity at 60 °C and pH 7.0. We optimized the conditions for bovine bone collagen hydrolysis and set up a process with high hydrolysis efficiency (99.4%) to prepare bovine bone collagen peptides, in which bovine bone collagen was hydrolyzed at 60 °C for 2 h with an enzyme–substrate ratio of 25 U/g. The hydrolysate contained 96.5% peptides that have a broad molecular weight distribution below 10000 Da. The hydrolysate showed good moisture-retention ability and a high hydroxyl radical (•OH) scavenging ratio of 73.2%, suggesting that the prepared collagen peptides have good antioxidative activity. Altogether, these results indicate that the thermophilic TLP A69 has promising potential in the preparation of bioactive collagen peptides, which may have potentials in cosmetics, food and pharmaceutical industries. This study lays a foundation for the high-valued utilization of bovine bone collagen.

Food Protein-Derived Antioxidant Peptides: Molecular Mechanism, Stability and Bioavailability

“…The modulation of the antioxidant defense system of the model organism by GLPGPM was highlighted, as demonstrated by the pretreatment of 0.1 mg/mL of the peptide that increased the activity of SOD and CAT (33.18% and 101.80%, respectively) and reduced ROS and MDA accumulation (17.89% and 48.90%, respectively). Another work evaluated the antioxidant capacity of enzymatic hydrolysates of bovine bone collagen [ 101 ]. The results pointed out the DPPH • (40.7%), • OH (31.8%), and O 2 •- (73.2%) scavenging activities of the hydrolysate (30 mg/mL).…”

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

“…SM9913 contained 60% oligopeptides with a molecular weight lower than 1000 Da, and exhibited good hydroxyl radical scavenging activity and promoted an effect on cell viability of human dermal fibroblasts [7]. A bovine bone collagen hydrolysate prepared with the thermolysin-like protease A69 from Anoxybacillus caldiproteolyticus 1A02591 contained 21.1% oligopeptides with a molecular weight lower than 1000 Da, and exhibited good moisture-retention ability and antioxidant activity [43]. The hydrolysate prepared from Bigeye tuna skin collagen contained peptides with molecular weights of 300-425 Da and had DPPH• scavenging activity [51].…”

“…With L-leucine as the standard, the content of free amino acids in the hydrolysate solution was determined by the ninhydrin method [62]. The content of peptides in the hydrolysate was calculated by subtracting the content of free amino acids from that of the hydrolysate in the solution [43]. The compositions of free and total amino acids of the hydrolysate were analyzed by using an amino acid analyzer HITACHI 835 (Tokyo, Japan).…”

Potential of the Liquid Fermentation of Fishery Waste by Paenibacillus elgii for Metalloprotease Production

“…P. elgii TKU051 protease reduced its activity with the appearance of Triton X-100 (53%) and was inhibited by ionic surfactants such as sodium dodecyl sulfate (SDS) (0%) and cetrimonium bromide (8%). Protease from the M4 family could be incorporated into detergent which effectively eliminated proteinaceous stains from textiles [18], therefore, we herein investigated the compatibility of P. elgii TKU051 protease with some commercial laundry detergents. Indeed, P. elgii TKU051 protease was compatible with two kinds of detergents (Ekos and Yeuhyang) with the residual activity of 94% and 93% (respectively) whereas, on Amah, the enzyme retained 63% of its initial activity.…”

“…Various microbes have been used to produce protease, for example, Bacillus [4][5][6], Paenibacillus [7][8][9], Aspergillus [10,11], Streptomyces [12,13], Pseudomonas [14,15], and Brevibacillus [16,17]. With a range of properties (e.g., thermal stability, alkaliphilic, substrate specificity, and chemical resistance), microbial proteases are used in a wide range of biotech applications such as the leather industry, pharmaceutical industry, detergent industry, food industry, and bioremediation [3,18]. Recently, the use of protease has gotten a lot of interest in producing bioactive protein hydrolysates as a possible alternative to chemical methods that may emit toxic chemicals or residual organic solvents [3,[19][20][21].…”