Mechanistic Insight Into the Production of Collagen Oligopeptides by the S8 Family Protease A4095

Abstract

Collagen oligopeptides have wide applications in foods, pharmaceuticals, cosmetics, and others due to their high bioactivities and bioavailability. The S8 family is the second-largest family of serine proteases. Several collagenolytic proteases from this family have been reported to have good potential in the preparation of collagen oligopeptides, however, the underlying mechanism remains unknown. A4095 was the most abundant S8 protease secreted by the protease-producing bacterium Anoxybacillus caldiproteolyticus 1A02591. Here, we characterized A4095 as an S8 collagenolytic protease and illustrated its structural basis to produce collagen oligopeptides. Protease A4095 preferentially hydrolyzed the Y-Gly peptide bonds in denatured bovine bone collagen, leading to high production (62.48% <1000 Da) of collagen oligopeptides. Structural and mutational analyses indicated that A4095 has a unique S1′ substrate-binding pocket to preferentially bind Gly, which is the structural determinant for the high production of collagen oligopeptides. This study provides mechanistic insight into the advantage of the S8 collagenolytic proteases in preparing collagen oligopeptides.

Production of marine bacterial metalloprotease A69 and evaluation of its potential in preparing soybean peptides with angiotensin‐converting enzyme‐inhibitory activity

“…SM9913, Aa2_1884 from Flocculibacter collagenilyticus SM1988 and A4095 from Anoxybacillus caldiproteolyticus 1A02591. [6][7][8] The oligopeptide contents in the collagen hydrolysates from these marine enzymes are generally higher than those from their terrestrial homologs, 8 suggesting the promising potential of these marine bacterial proteases in collagen peptides preparation. In addition to these proteases, there may be many other marine bacterial proteases suitable for preparation of bioactive peptides, although this awaits exploration.…”

“…Several serine proteases of the MEROPS S8 family from marine bacteria have been used in collagen peptides preparation, including MCP‐01 from Pseudoaltermonas sp. SM9913, Aa2_1884 from Flocculibacter collagenilyticus SM1988 and A4095 from Anoxybacillus caldiproteolyticus 1A02591 6‐8 . The oligopeptide contents in the collagen hydrolysates from these marine enzymes are generally higher than those from their terrestrial homologs, 8 suggesting the promising potential of these marine bacterial proteases in collagen peptides preparation.…”