Solid state fermentation is a promising technology largely used in biotechnology process and is a suitable strategy for producing low-cost enzymatic products. At the present study, a novel enzyme obtained through solid state fermentation using Aspergillus sydowii was herein purified and characterized. The fermentations used coffee ground residue as substrate and the crude enzyme was submitted through further purification steps of: acetonic precipitation, DEAE-Sephadex and Superdex G-75 column. Both crude and purified enzymes were submitted to biochemical characterization of their thermostability, optimal temperature and pH, effects of inhibitors and metal ions. A purified protease was obtained with yield of 5.9-fold and 53% recovery, with maximal proteolytic activity of 352.0 U/mL. SDS-PAGE revealed a band of protein at 47.0 kDa. The enzyme activity was abolished in the presence of phenyl-methyl sulfonyl fluoride and partially inhibited against Triton X-100 (78.0%). The optimal activity was found in pH 8.0 at 45°C of temperature. Besides, the enzyme showed stability between 35°C and 50°C. It was possible to determine appropriate conditions to the obtainment of thermostable proteases with biotechnological interest associated with a method that concomitantly shows excellent production levels and recovery waste raw material in a very profitable process.

Fibrinolytic proteases operate directly on fibrin clot and are able to maintain blood flow. Fungi show up as viable sources for obtaining this enzyme. The purpose of this systematic review is to is to unveil all the information concerning production, purification and characterization of fibrinolytic proteases by fungi. The search was conducted in ScienceDirect, PubMed and Scopus databases, using as keywords “(Fibrinolytic enzyme) OR (Fibrinolytic protease) AND (Fungal or Fungus or Fungi)”. Delimiting period of 10 years (2011-2021). The results obtained were filtered by selection criteria, and review articles or articles outside the scope of this work were excluded. The articles were evaluated and scored (0-10) according to pre-established criteria. None of the studies obtained the score 10, however the study with the highest score (9) presented relevant data in all criteria analyzed, obtaining fibrinolytic enzyme from Xylaria curta. Among the 21 selected articles, 12 different genera appear and Submerged Fermentation and purification of Serino-proteases were more described. This work also observed a greater representation of purification and characterization steps, indicating the need for attention to cultivation process and enzymatic application. It is clear that the production of these enzymes by fungi is pertinent towards the high recovery observed even after purification and its tendency for pharmaceutical application.

Spent coffee grounds (SCG) are a widely available agro-industrial residue rich in carbon and phenolic compounds, presenting significant potential for biotechnological valorization. This study evaluated the use of SCG as a suitable substrate for fungal laccase production and the application of the resulting fermented biomass (RFB), a mixture of fermented SCG and fungal biomass as a biosorbent for textile dye removal. Two fungal strains, namely Lentinus crinitus UCP 1206 and Trametes sp. UCP 1244, were evaluated in both submerged (SmF) and solid-state fermentation (SSF) using SCG. L. crinitus showed superior performance in SSF, reaching 14.62 U/g of laccase activity. Factorial design revealed that a lower SCG amount (5 g) and higher moisture (80%) and temperature (30 °C ± 0.2) favored enzyme production. Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) analyses confirmed significant structural degradation of SCG after fermentation, especially in SSF. Furthermore, SCG and RFB were chemically activated and evaluated as biosorbents. The activated carbon from SCG (ACSCG) and RFB (ACRFB) exhibited high removal efficiencies for Remazol dyes, comparable to commercial activated carbon. These findings highlight the potential of SCG as a low-cost, sustainable resource for enzyme production and wastewater treatment, contributing to circular bioeconomy strategies.

Fructooligosaccharides (FOS) are prebiotics of interest to the food industry. These compounds can be produced through the transfructosylation reaction by the enzyme fructofuranosidase. This enzyme is widely produced by fungi in a medium rich in sugar. Therefore, in this work, the main objectives were production, purification, biochemical characterization of a novel fructofuranosidase enzyme by Penicillium citreonigrum URM 4459 and synthesize and evaluate the antibacterial potential of fructooligosaccharides. With respect to sucrose hydrolysis, the optimal pH was 5.5, the apparent Km for purified FFase was 3.8 mM, the molecular mass was 43.0 kDa, estimated by gel filtration on Superdex increase G75 controlled by AKTA Avant 25 and confirmed by 10% SDS-PAGE under denaturing condition. Also, the isoelectric point was 4.9. The fractions obtained with enzymatic activities, both stable at acidic pH and high temperatures, as well as being able to produce FOS. Regarding antibacterial activity, the FOS produced in this study showed better results than commercial FOS and other carbon sources. Thus, this work presents relevant data for the use of P. citreonigum to produce fructofuranosidase and consequently FOS and can be used in the food and pharmaceutical industry.

(1) Background: Polysaccharide films are promising vehicles for the delivery of bioactive agents such as collagenases, as they provide controlled release at the wound site, facilitating tissue regeneration. This study aimed to investigate the physicochemical properties of Cassia grandis polysaccharide films with immobilized collagenase from Streptomyces parvulus (DPUA/1573). (2) Methods: Galactomannan was extracted from Cassia grandis seeds for film production with 0.8% (w/v) galactomannan and 0.2% (v/v) glycerol with or without collagenases. The films underwent physical-chemical analyses: Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), thermogravimetric analysis (TGA), color and opacity (luminosity-L*, green to red-a*, yellow to blue-b*, opacity-Y%), moisture content, water vapor permeability (WVP), thickness, contact angle, and mechanical properties. (3) Results: The results showed similar FTIR spectra to the literature, indicating carbonyl functional groups. Immobilizing bioactive compounds increased surface roughness observed in SEM. TGA indicated a better viability for films with immobilized S. parvulus enzymes. Both collagenase-containing and control films exhibited a bright-yellowish color with slight opacity (Y%). Mechanical tests revealed decreased rigidity in PCF (−25%) and SCF (−41%) and increased deformability in films with the immobilized bioactive compounds, PCF (234%) and SCF (295%). (4) Conclusions: Polysaccharide-based films are promising biomaterials for controlled composition, biocompatibility, biodegradability, and wound healing, with a potential in pharmacological applications.

Collagenases are enzymes with broad biotechnological applications in medicine. This study describes the production and characterization of a collagenase from Aspergillus serratalhadensis URM 7866, isolated from the Caatinga biome. Solid-state fermentations were conducted using wheat bran under varying conditions of pH (6, 7, 8), moisture content (50%, 60%, 70%), and substrate concentration (2.5 g, 5 g, 10 g). The optimal condition—10 g of wheat bran at pH 8 and 70% moisture—yielded the highest collagenolytic activity (177.96 U/mL) and a specific activity of 50.55 U/mg. The enzyme was purified via multiple chromatography, with pre-purification and final purification factors of 18.09 and 20.21, respectively, reaching a specific activity of 1021.86 U/mg. The enzyme showed optimal activity at 50 °C and pH 8, with stability from 20 to 40 °C and pH 7–9. PMSF caused >80% inhibition; EDTA caused ~34% inhibition. Activity increased with Na⁺ and Ca²⁺ and was inhibited by Zn²⁺. The enzyme retained full activity in anionic and non-ionic surfactants (1–10%). FTIR confirmed characteristic amide bands, and kinetic analysis revealed a Km of 1.72 mg/mL and Vmax of 6.89 mg/mL/min. These findings support its potential for alkaline and surfactant-rich industrial processes.