Have already been purified from other Basidiomycota and Ascomycota species including Coprinellus radians (CraUPO) (Anh et al., 2007), Marasmius rotula (MroUPO) (Gr e et al., 2011), and Chaetomium globosum (CglUPO) (Kiebist et al., 2017), which is indicative for their widespread occurrence within the fungal kingdom. As well as these wild (i.e., 5-HT5 Receptor Antagonist web nonrecombinant) enzymes, you can find other UPOs, e.g., fromCoprinopsis cinerea (rCciUPO) (Babot et al., 2013) and Humicola insolens (rHinUPO) (Kiebist et al., 2017), which might be only known as recombinant proteins heterologously expressed by Novozymes A/S (Bagsvaerd, Denmark) in the mold Aspergillus oryzae (Landvick et al., 2016), and very lately further UPOs heterologously expressed in Escherichia coli (Linde et al., 2020). Initially, UPO PAK3 manufacturer enzymes had been shown to catalyze oxygenation reactions on aromatic compounds (Hofrichter et al., 2010), and their action on aliphatic compounds was demonstrated later (Guti rez et al., 2011; Peter et al., 2011). Right here, we demonstrate a promising enzymatic technologies to epoxidize, under mild and environmentally friendly conditions, complex mixtures of no cost and methylated fatty acids from representative vegetable oils, which had been previously applied on isolated pure fatty acids (Aranda et al., 2018), for its industrial application within the production of biobased binder ingredients, in collaboration with interested organizations. This contains the usage of two wild UPOs, namely MroUPO and CglUPO, and recombinant rHinUPO, all of them with preferential epoxidation (vs. hydroxylation) oxygenation patterns. These and associated fungal peroxygenases elude a number of the limitations of other monooxygenases given that they are secreted proteins, thus much more stable, and only call for H2 O2 for activation (Wang et al., 2017; Hofrichter et al., 2020). In addition, their recent expression as soluble and active enzymes in Escherichia coli is expanding the amount of UPO enzymes out there from associated genes in sequenced genomes (Linde et al., 2020) and, simultaneously, enabling the rational style in the available UPOs as ad hoc biocatalysts of industrial interest, employing the protein engineering tools (Carro et al., 2019; Gonz ez-Benjumea et al., 2020; Municoy et al., 2020).Materials AND Techniques OilsFour refined vegetable oils–namely rapeseed, soybean, sunflower, and linseed oils–were supplied by the Cargill business and stored at four C, prior to their saponification, transesterification and use as UPO substrates. For characterization from the complete lipid profiles (“intact” lipids), aliquots had been straight treated with BSTFA [N,Obis-(trimethylsilyl)trifluoroacetamide] at 80 C for 1 h, and analyzed by GC-MS.EnzymesMroUPO and CglUPO are wild enzymes isolated at JenaBios (Jena, Germany) from pure cultures of M. rotula DSM 25031 and C. globosum DSM 62110 from the German Collection of Microorganisms and Cell Cultures (Braunschweig, Germany). rHinUPO is often a recombinant enzyme obtained at Novozymes A/S (Bagsvaerd, Denmark) (Kiebist et al., 2017), by heterologous expression with the cloned gene in the A. oryzae industrial host, employing proprietary technology (Landvick et al., 2016). In all cases, the secreted enzyme was recovered after eliminating the fungal mycelium by filtration of liquid cultures, concentrated by ultrafiltration or ammonium sulfate precipitation, and purifiedFrontiers in Bioengineering and Biotechnology | www.frontiersin.orgJanuary 2021 | Volume 8 | ArticleGonz ez-Benjumea et al.Biobased Epoxides by Funga.
