Specific chemical incorporation of l-DOPA and functionalized l-DOPA-hyaluronic acid in Candida antarctica lipase: creating potential mussel-inspired bioadhesives

Abstract

Mussel adhesives proteins have been extensively studied as a promising alternative in bioadhesives due to their ability to provide durable anchoring under different surfaces in seawater. These charasteristics have been attributed to the presence of the reduced catechol form, 3,4-dihydroxyphenylalanine (DOPA) of its structure. However, its practical applications have been limited due to drawbacks with natural extraction. Here, a novel method have been described for site-specific chemical incorporation of l-3,4-dihydroxyphenylalanine methyl ester (l-DOPAME) into proteins, in particular Candida antartica fraction B (CAL-B) lipase. Two strategies were followed, direct conjugation of DOPA at the C-terminus on the surface of the protein, and protein conjugation with tailor-made glycopolymers (DOPA-hyaluronic acid (HA) polymers) at the N-terminus. In all cases, the characterization of the new DOPA-proteins was carried out using circular dichroism, fluorescence or mass spectrometry. An improvement in the activity (in some cases more than 2 times) or the thermostability of CAL-B (with a half live 4 fold greater in some cases) was found by the incorporation of DOPA molecules. These DOPA-proteins showed excellent underwater covalent adhesive ability on amino functionalized surfaces in aqueous media compared to other modified [e.g. tyrosine modified (TYR)] CAL-B proteins. At pH 8.5, CALB-DOPA proteins were completely adsorbed after 90 min of incubation, whereas about 10% of CALB-HA or CALB-TYR proteins were adsorbed at the same time. Native CAL-B adsorption was not observed. These results suggest a potential application of these DOPA-proteins as bioglues or bioadhesives for practical underwater applications.

Bio-Based Adhesives for Orthopedic Applications: Sources, Preparation, Characterization, Challenges, and Future Perspectives

“…Based on biomechanical studies, it is known that these bio-adhesive polymers have better strength of bio-adhesion when compared to fibrin glue, but they also have a poorer strength of bio-adhesion than cyanoacrylates on bovine cortical bone specimens. Excellent biocompatibility was also demonstrated in in vitro cell testing, so this bio-based adhesive can be a promising candidate for clinical utilization [21,42,[46][47][48].…”

Synthesis of silver and gold nanoparticles–enzyme–polymer conjugate hybrids as dual-activity catalysts for chemoenzymatic cascade reactions

“…35 These polymers have been recently used for stabilization of multimeric enzymes, 19 and also, we observed important influences on the selectivity and specificity of enzymes after their modification. 18,36,37 Dextrans with molecular weights of 6 kDa and 2000 kDa, respectively, were modified firstly using sodium periodate to create dextranaldehyde (Dext-Asp). The chemical incorporation of aspartic acid molecules into dextran-aldehyde polymer resulted in the final dextran-aspartic acid polymers called Dext6kDa and Dext2000kDa.…”

“…14 Recent studies have demonstrated that the physical or chemical modification of enzymes generates novel bioconjugates with improved catalytic properties. [18][19][20][21] This is a very important issue, in particular, when a cascade process is developed. These novel nanobiohybrids could have a pivotal advantage over other systems since they are able to exhibit an enzymatic and metallic activity in the same compartment.…”

Pd-Oxazolone complexes conjugated to an engineered enzyme: improving fluorescence and catalytic properties

“…Enzyme lid oligopeptide (orange). egy recently reported 31 by us using N-acetyl-tyrosine (Ac-Tyr) as molecule (Fig. S6 †).…”

“…of NHS (37.46 mg) were added in 5 mL of H 2 O adjusted to pH 4.8-5.2. 31 This solution was allowed to react for 1 h. Then, 0.5 mL (50 eq.) of the previous solution was added to 3.5 mL of H 2 O adjusted to pH = 7 and 0.5 g of GTL or GTL-C114 immobilized on butyl-Sepharose protein at r.t., and the mixture was left to react overnight.…”