Combi-lipase for heterogeneous substrates: a new approach for hydrolysis of soybean oil using mixtures of biocatalysts

Abstract

The concept of the combi-lipase biocatalyst has been proposed. It is based on the combination of different lipases as biocatalysts in reactions using heterogeneous substrates.

Current status and new developments of biodiesel production using fungal lipases

“…This concept could be better explored for the production of biodiesel, since vegetable oils, which are the main raw materials for biodiesel synthesis, are heterogeneous substrates, containing triglycerides formed by very different fatty acids, glycerides with lower substitutions, and even free fatty acids [133,134]. Poppe et al [134] evaluated enzymatic ethanolysis of olive and palm oils using mixtures of three immobilized lipases as biocatalysts: Novozym 435 (CALB), Lipozyme TL-IM (TLL), and Lipozyme RM-IM (RML).…”

Optimization of ethyl ester production from olive and palm oils using mixtures of immobilized lipases

“…The concept is based on the fact that a composed biocatalyst of a mixture of different lipases should be more effective on heterogeneous substrates than one specific lipase [17]. It was applied for the hydrolysis of soybean oil, with results showing that the mixture of lipases from Rhizomucor miehie (RML) and Candida antarctica (CALB) was more efficient than the lipase from Thermomyces lanuginosus (TLL), which was the most active enzyme, when used individually [16].…”

Moving towards a Competitive Fully Enzymatic Biodiesel Process

“…In this case, best immobilization was obtained on polypropylene supports: Accurel EP100 for LipA and Accurel MP1000 for LipC and LipCmut. Lip I.3, requiring a refolding step [21], was poorly immobilized on all supports tested (best results for Accurel MP1000). The behavior of all four immobilized lipases was tested for triolein transesterification, the best results being obtained for lipases immobilized on Accurel MP1000.…”

“…Moreover, lipases can perform reactions with high specificity, regioselectivity and enantioselectivity, becoming the most used enzymes in synthetic organic chemistry [13,[18][19][20]. Therefore, there is substantial interest in developing appropriate lipase preparations for their use in biodiesel synthesis [21,22]. As an initial approach, the study of biodiversity is a valuable tool for the discovery of new lipolytic enzymes useful for biocatalysis [20,23,24].…”

“…On the contrary, simple enzyme adsorption allows preparation and use of immobilized lipases under mild conditions without significant loss of activity [46], and the associated process is very simple, with the carrier being easily recovered for repeated immobilization rounds [22,46]. Therefore, four non-commercial lipases from Pseudomonas species-LipA and LipC from P. aeruginosa 42A2 [31], LipCmut, the thermostable mutant of LipC previously described [47], and LipI.3 from Pseudomonas CR611 [21], which were produced in either homologous or heterologous hosts-were prepared, immobilized on different cheap supports, and examined for potential biodiesel production. The enzymes were obtained directly from crude cell extracts or from growth culture supernatants, and further immobilized on different carriers: Polypropylene polymers like Accurel EP100 or Accurel MP1000 [48,49] and a silica powder, Celite ® 545 [50] by simple adsorption.…”