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Kinetic Analysis of the Digestion of Bovine Type I Collagen Telopeptides with Porcine Pepsin
Author(s) -
Qian Jun,
Okada Yukari,
Ogura Takayuki,
Tanaka Keisuke,
Hattori Shunji,
Ito Shinji,
Satoh Junko,
Takita Teisuke,
Yasukawa Kiyoshi
Publication year - 2016
Publication title -
journal of food science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.772
H-Index - 150
eISSN - 1750-3841
pISSN - 0022-1147
DOI - 10.1111/1750-3841.13179
Subject(s) - pepsin , chemistry , type i collagen , digestion (alchemy) , cleavage (geology) , chromatography , enzyme , biochemistry , materials science , biology , fracture (geology) , composite material , endocrinology
Abstract Collagen is frequently digested using pepsin in industries to produce a triple helical collagen without the N‐ and C‐terminal telopeptides. However, kinetic analysis of this reaction is difficult because several Lys residues in the N‐ and in the C‐terminal telopeptides form covalent bonds, leading to multiple substrates species, and pepsin cleaves collagen at various sites in the N‐terminal and in the C‐terminal telopeptides, yielding different products. Here we performed kinetic analysis of the digestion of bovine type I collagen with porcine pepsin. The reaction could be monitored by SDS‐PAGE by measuring the intensity of the protein bands corresponding to the variant β11 chain. We obtained kinetic parameters relative to the decrease in the variant β11 chain upon digestion. At pH 4.0, the K m and k cat values increased with increasing temperature (30 to 65 °C), although the k cat / K m values were stable. Additional cleavage at the helical region was detected at 45 to 65 °C. At 37 °C, the K m and k cat values increased with decreasing pH, and the k cat / K m values at pH 2.1 to 4.5 were stable and higher than those at pH 5.0 and 5.5. No additional cleavage was detected at the examined pH. Thus, the optimal pH and temperatures for selective digestion of collagen telopeptides with pepsin are 2.1 to 4.5 and 30 to 40 °C, respectively. These results suggest that the method might be useful for the kinetic analysis of the digestion of collagen telopeptides with pepsin.