Loosenin-Like Proteins from Phanerochaete carnosa Impact Both Cellulose and Chitin Fiber Networks.

Microbial expansin-related proteins are ubiquitous across bacterial and fungal organisms and reportedly play a role in the modification and deconstruction of cell wall polysaccharides, including lignocellulose. So far, very few microbial expansin-related proteins, including loosenins and loosenin-like (LOOL) proteins, have been functionally characterized. Herein, four LOOLs encoded by Phanerochaete carnosa and belonging to different subfamilies (i.e., PcaLOOL7 and PcaLOOL9 from subfamily A and PcaLOOL2 and PcaLOOL12 from subfamily B) were recombinantly produced and the purified proteins were characterized using diverse cellulose and chitin substrates. The purified PcaLOOLs weakened cellulose filter paper and cellulose nanofibril networks (CNF); however, none significantly boosted cellulase activity on the selected cellulose substrates (Avicel and Whatman paper). Although fusing the family 63 carbohydrate-binding module (CBM63) of BsEXLX1 encoded by Bacillus subtilis to PcaLOOLs increased their binding to cellulose, the CBM63 fusion appeared to reduce the cellulose filter paper weakening observed using wild-type proteins. Binding of PcaLOOLs to alpha-chitin was considerably higher than that to cellulose (Avicel) and was pH dependent, with the highest binding at pH 5.0. Amendment of certain PcaLOOLs in fungal liquid cultivations also impacted the density of the cultivated mycelia. The present study reveals the potential of fungal expansin-related proteins to impact both cellulose and chitin networks and points to a possible biological role in fungal cell wall processing. IMPORTANCE The present study deepens investigations of microbial expansin-related proteins and their applied significance by (i) reporting a detailed comparison of diverse loosenins encoded by the same organism, (ii) considering both cellulosic and chitin-containing materials as targeted substrates, and (iii) investigating the impact of the C-terminal carbohydrate binding module (CBM) present in other expansin-related proteins on loosenin function. By revealing the potential of fungal loosenins to impact both cellulose and chitin-containing networks, our study reveals a possible biological and applied role of loosenins in fungal cell wall processing.

Semisynthetic transformation of banana peel to enhance the conversion of sugars to 5-hydroxymethylfurfural.

This study aimed to efficiently convert banana peels (BP) into 5-hydroxymethylfurfural (HMF) by using an integrated mechanoenzymatic/catalytic approach. There is no report on HMF production using mechanoenzymatic hydrolysis. Moreover, this method enables saccharification of lignocellulose without bulk solvents or pretreatment. The effects of the reaction volume, milling time, and reactive aging (RAging) on the mechanoenzymatic hydrolysis of BP were studied. The solvent-free enzymatic hydrolysis of BP under RAging conditions was found to provide higher glucose (40.5 wt%) and fructose (17.2 wt%) yields than chemical hydrolysis. Next, the conversion of the resulting monosaccharides into HMF in the presence of the AlCl3·H2O/HCl-DMSO/H2O system resulted in 71.9 mol% yield, which is so far the highest HMF yield obtained from cellulosic food wastes. Under identical reaction conditions, direct conversion of untreated BP to HMF yielded 22.7 mol% HMF, suggesting that mechanoenzymatic hydrolysis greatly promotes the release of sugars from BP to improve HMF yield.