Immobilisation on mesoporous silica and solvent rinsing improve the transesterification abilities of feruloyl esterases from Myceliophthora thermophila

Abstract

The immobilisation of four feruloyl esterases (FAEs) (FaeA1, FaeA2, FaeB1, FaeB2) from the thermophilic fungus Myceliophthora thermophila C1 was studied and optimised via physical adsorption onto various mesoporous silica particles with pore diameters varying from 6.6nm to 10.9nm. Using crude enzyme preparations, enrichment of immobilised FAEs was observed, depending on pore diameter and protein size. The immobilised enzymes were successfully used for the synthesis of butyl ferulate through transesterification of methyl ferulate with 1-butanol. Although the highest butyl ferulate yields were obtained with free enzyme, the synthesis-to-hydrolysis ratio was higher when using immobilised enzymes. Over 90% of the initial activity was observed in a reusability experiment after nine reaction cycles, each lasting 24h. Rinsing with solvent to remove water from the immobilised enzymes further improved their activity. This study demonstrates the suitability of immobilised crude enzyme preparations in the development of biocatalysts for esterification reactions.

Release of phenolic acids from sunflower and rapeseed meals using different carboxylic esters hydrolases from Aspergillus niger

“…Therefore, given the presence of significant amounts of glucose (Naczk &Shahidi, 1990), as well as mono-and disinapoyl glucose isomers in the NI-RSM extract (Laguna et al, 2018), it can be reasonably assumed that this new compound resulted from transesterification of sinapine with the aforementioned molecules extracted from the meal. If so, these results would confirm the well-known acyl transferase activity of AnFaeA (Hatzakis and Smonou, 2005;Hüttner et al, 2017;Vafiadi et al, 2009). However, the reason why such a reaction only occurred in the NI-RSM methanolic extract remains to elucidate.…”

Glycosylation influences activity, stability and immobilization of the feruloyl esterase 1a from Myceliophthora thermophila

“…The production of Mt Fae1a in M. thermophila C1 was done as previously reported by over-production in a low background production host strain (Visser et al 2011). The biomass-free fermentation broth was concentrated and dialyzed, and the resulting crude FAE extract was freeze-dried until use [as described for FaeB2 in (Hüttner et al 2017)]. M. thermophila is also known as Chrysosporium lucknowense and Thermothelomyces thermophila (Marin-Felix et al 2015).…”

Feruloyl esterase immobilization in mesoporous silica particles and characterization in hydrolysis and transesterification

“…Influence of the pH of the immobilization buffer on the enzyme loading [ 24 ] and selectivity [ 7 ], has been investigated previously for FoFaeC, resulting in an increase of selectivity for transesterification. In another study, using four different FAEs from the thermophilic fungus Myceliophtora thermophila C1 and 10.1 nm pore size SBA-15, the selectivity towards transesterification was decreased for three out of four enzymes [ 28 ]. In the present study, using a commercial FAE, immobilization led to a decreased selectivity for transesterification (Fig.…”

“…In the present study, enzymes immobilized on MPS were dried before being used in transesterification reactions. One way to improve the BFA yield of immobilized enzymes might be to replace the drying step by solvent rinsing as it has been shown to improve the productivity of a FAE from Myceliophtora thermophila C1 from 3 to 5 mmol BFA /g enz /h [ 28 ].…”