Enzymes offer interesting features as biological catalysts for industry: high specificity, activity under mild conditions, accessibility, and environmental friendliness. Being able to produce enzymes in large quantities and having them available in a stable and reusable form reduces the production costs of any enzyme-based process. Agricultural residues have recently demonstrated their potential as substrates to produce ligninolytic enzymes by different white rot fungi. In this study, the biotechnological production of a manganese peroxidase (MnP) by Irpex lacteus was conducted through solid-state fermentation (SSF) with wheat straw as substrate and submerged fermentation (SmF) employing wheat straw extract (WSE). The obtained enzyme cocktail also showed manganese-independent activity (MiP), related to the presence of a short MnP and a dye-decolorizing peroxidase (DyP) which was confirmed by shotgun proteomic analyses. In view of the enhanced production of ligninolytic enzymes in SmF, different parameters such as WSE concentration and nitrogen source were evaluated. The highest enzyme titers were obtained with a medium formulated with glucose and peptone (339 U/L MnP and 15 U/L MiP). The scale-up to a 30 L reactor achieved similar activities, demonstrating the feasibility of enzyme production from the residual substrate at different production scales. Degradation of five emerging pollutants was performed to demonstrate the high oxidative capacity of the enzyme. Complete removal of hormones and bisphenol A was achieved in less than 1 h, whereas almost 30% degradation of carbamazepine was achieved in 24 h, which is a significant improvement compared to previous enzymatic treatments of this compound. Key points: • Wheat straw extract is suitable for the growth of I. lacteus. • The enzyme cocktail obtained allows the degradation of emerging contaminants. • Mn-dependent and Mn-independent activities increases the catalytic potential. Graphical

Interest in the development of biorefineries and biotechnological processes based on renewable resources has multiplied in recent years. This driving force is the result of the availability of lignocellulosic biomass and the range of applications that arise from its use and valorization. The approach of second-generation sugars from lignocellulosic biomass opens up the possibility of producing biotechnological products such as enzymes as a feasible alternative in the framework of biorefineries. It is in this context that this manuscript is framed, focusing on the modelling of a large-scale fermentative biotechnological process to produce the enzyme manganese peroxidase (MnP) by the fungus Irpex lacteus using wheat straw as a carbon source. The production scheme is based on the sequence of four stages: pretreatment of wheat straw, seed fermenters, enzyme production and downstream processes. For its environmental assessment, the Life Cycle Assessment methodology, which allows the identification and quantification of environmental impacts associated with the process, was utilized. As the main finding, the stages of the process with the highest environmental burdens are those of pretreatment and fermentation, mainly due to energy requirements. With the aim of proposing improvement scenarios, sensitivity analyses were developed around the identified hotspots. An improvement in the efficiency of steam consumption leads to a reduction of environmental damage of up to 30%.

Unspecific peroxygenase (UPO) presents a wide range of biotechnological applications. This study targets the use of by-products from bioethanol synthesis to produce UPO by Agrocybe aegerita. Solid-state and submerged fermentations (SSF and SmF) were evaluated, achieving the highest titers of UPO and laccase in SmF using vinasse as nutrients source. Optimized UPO production of 331 U/L was achieved in 50% (v:v) vinasse with an inoculum grown for 14 days. These conditions were scaled-up to a 4 L reactor, achieving a UPO activity of 265 U/L. Fungal proteome expression was analyzed before and after UPO activity appeared by shotgun mass spectrometry proteomics. Laccase, dye-decolorizing peroxidases (DyP), lectins and proteins involved in reactive oxygen species (ROS) production and control were detected (in addition to UPO). Interestingly, the metabolism of complex sugars and nitrogen sources had a different activity at the beginning and end of the submerged fermentation.