Cross-linking of collagen with laccases and tyrosinases

• Published in: Materials Science & Engineering C Vol. 31; no. 5; pp. 1068 - 1077
• Main Authors: Jus, S., Stachel, I., Schloegl, W., Pretzler, M., Friess, W., Meyer, M., Birner-Gruenberger, R., Guebitz, G.M.
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 20.07.2011
• Subjects: Collagen; Collagens; Compressive strength; Crosslinking; Degradation; Enzymes; Laccase; Microorganisms; Tyrosinase; Crosslinking; Collagen; Tyrosinase; Laccase
• ISSN: 0928-4931; 1873-0191
• DOI: 10.1016/j.msec.2011.03.007

Oxidation of acid soluble collagen (ASC), collagen suspension and BrCN-peptides (BrCN-P) with tyrosinases from B. obtusa (BoT1, BoT2) and A. bisporus (AbT) and laccases from T. versicolor (TvL) and T. hirsuta (ThL) resulted in UV/VIS peaks at 475 nm and 305 nm indicating formation of reactive o-quinones and cross-linked components. Concomitant oxygen consumption was higher for the low molecular weight enzymes (TvL and BoT2) indicating limited accessibility. SDS-PAGE and SEC bands at higher MW demonstrated the formation of cross-linked material. LC-MS/MS analysis suggested the involvement of tyrosine residues in cross-linking without major changes of sequence similarities to untreated collagen. However, an increase of the SEC α-peak together with a decrease of β-peak and the 1235/1450 cm − 1 ratio (FTIR) indicated partial degradation. Crosslinking was enhanced by phenolic molecules such as catechine which lead to increased denaturation temperature and reduced degradation by microbial collagenase. The tensile strength was increased whereas resistance to compressive forces was not influenced. [Display omitted] ► Enzymatic cross-linking of collagen with oxidoreductases demonstrated. ► Oxidation of collagen by laccase and tyrosinase proven. ► Applications potential of enzymatic collagen cross-linking in medical applications shown.