Chemically Modified Lipase from Thermomyces lanuginosus with Enhanced Esterification and Transesterification Activities

Abstract

Lipase from Thermomyces lanuginosus is one of the most explored enzymes for the esterification of several added-value industrial compounds, such as biodiesel, fragrances, and flavors. Its selectivity in these reactions is mostly related with its activity towards small alcohols. In this work, the impact of the chemical modification, with 4 dodecyl chains at its surface, was evaluated regarding its transesterification and esterification activities, comparing with the native form. Linear size-differentiated alcohols (from 1 to 20 carbons in the aliphatic chain) were used to explore for the first time the effect of the chain length in both transesterification and esterification reactions, using p-nitrophenyl palmitate and oleic acid as model compounds, respectively. The chemically modified lipase showed an outstanding improvement of its catalytic performance than the native enzyme, being this increase directly proportional to the size of the alcohols chain used as substrates. The enormous potential and remarkable versatility of this novel super catalyst was here demonstrated, where diverse types of esters, differing in their potential applications (biodiesel, cosmetics, fine chemistry), were efficiently synthesized. The produced esters were fully characterized by 1 H NMR, GC-MS, and FTIR.

Characteristics of Crosslinking Polymers Play Major Roles in Improving the Stability and Catalytic Properties of Immobilized Thermomyces lanuginosus Lipase

“…Compared with butanol, the initial esterification rate between oleic acid and ethanol was much faster for all lipase preparations. Noro et al [ 31 ] found that native free TLL showed higher affinity toward alcohols with three or less carbons than those with a longer chain length during the esterification of oleic acid with various alcohols. However, the reverse reaction happened simultaneously during the esterification between oleic acid and ethanol, as an even lower amount of ester was detected at 2 h than that at 0.5 h, especially for free lipase.…”

Solvent‐free synthesis of hydrophobic and amphiphilic esters using a chemically modified lipase from Thermomyces lanuginosus: a comparative study with native and immobilized forms

“…In the first approach (Figure 2B), the compounds were synthesized with native LTL and chemically modified LTL (cmLTL) in a solvent-reactional system, resembling its traditional reaction conditions. [8,17] The data revealed that native TL showed limited activity for the tested compounds, demonstrating only residual activity for esters synthesized from heptanol with heptanoic, octanoic and decanoic acids. When decanol was used as the alcohol source, the native lipase was completely inefficient.…”

“…[14] The activity improvement was not only observed for hydrolysis reactions but also for esterification and transesterification purposes. [8] Apart from increasing enzyme catalytic activity, the chemical modification strategy, compared to physical immobilization onto solid supports, is often a more cost-effective option for industrial-scale applications. Furthermore, the chemical modification technique provides a way to reduce the dependency on organic solvents while increasing enzyme performance.…”

“…[4,5] Therefore, biocatalysis provides an appealing option for industries seeking sustainable, efficient, and environmentally friendly processes. However, most approaches applied industrially still require the use of an organic solvent, such as dimethyl sulfoxide, dimethylformamide, [6] n-hexane, [7] or n-heptane, [8] to enhance the enzymes' mobility and productivity, and substrate solubility. Several drawbacks are associated to these solvents including high cost, potential toxicity, and environmental concerns.…”

Chemical modification of lipases: A powerful tool for activity improvement

“…Later, the authors studied the transesterification ( p ‐nitrophenyl palmitate) and esterification (oleic acid) activity of the modified lipase against different chain‐length alcohols. [ 68 ] Again, the lipase grafted with four dodecyl chains, compared to the native enzyme, showed remarkable catalytic activity for synthesis reactions. The increase in activity was more pronounced the longer the alcohol chain was used.…”