Recent advances in the intellectual property landscape of filamentous fungi.

For centuries, filamentous fungi have been used in the making of food and beverages, and for decades for the production of enzymes and pharmaceuticals. In the last decades, the intellectual property (IP) landscape for fungal technology has seen an ever increasing upward trend, introducing new and promising applications utilising fungi. In this review, we highlight fungi-related patent applications published during the last 5 years (2015-2020), identify the key players in each field, and analyse future trends. New developments in the field of fungal technology include the increased use of filamentous fungi as a food source (mycoprotein), using fungi as biodegradable materials, in wastewater treatment, in integrated biorefineries and as biological pest agents. Biotechnology companies in Europe and the US are currently leading when it comes to the number of patents in these areas, but Asian companies and research institutes, in particular in China, are becoming increasingly important players, for example in pesticide formulation and agricultural practices.

Effect of single nanoparticle-nanopore interaction strength on ionic current modulation.

Solid-state nanopores are rapidly emerging as promising platforms for developing various single molecule sensing applications. The modulation of ionic current through the pore due to translocation of the target molecule has been the dominant measurement modality in nanopore sensors. Here, we focus on the dwell time, which is the duration taken by the target molecule or particle to traverse the pore and study its dependence on the strength of interaction of the target with the pore using single gold nanoparticles (NPs) as targets interacting with a silicon nitride (SiN) nanopore. The strength of interaction, which in our case is electrostatic in nature, can be controlled by coating the nanoparticles with charged polymers. We report on an operating regime of this nanopore sensor, characterized by attractive interactions between the nanoparticle and the pore, where the dwell time is exponentially sensitive to the target-pore interaction. We used negatively and positively charged gold nanoparticles to control the strength of their interaction with the Silicon Nitride pore which is negatively charged. Our experiments revealed how this modulation of the electrostatic force greatly affects the dwell time. Positively charged NPs with strong attractive interactions with the pore resulted in increase of dwell times by 2-3 orders of magnitude, from 0.4 ms to 75.3 ms. This extreme sensitivity of the dwell time on the strength of interaction between a target and nanopore can be exploited in emerging nanopore sensor applications.

Conversion of food and kitchen waste to value-added products.

Food and kitchen waste - omnipresent in every corner of the world serve as an excellent source of value added products owing to high organic content. Regardless of existence of various traditional methods of land filling or biogas production used to harness food waste energy, effective conversion of food to valuable resources is often challenged by its heterogenous nature and high moisture content. The current paper tries to lay down the prospects and consequences associated with food waste management. The various social, economical and environmental concerns associated with food waste management especially in terms of green house gas emission and extended rate of leachate generation also has been discussed. The difficulties in proper collection, storage and bioconversion of food waste to valuable by-products are pointed as a big hurdle in proper waste management. Finally, the wide array of value added products developed from food waste after pretreatment are also enlisted to emphasis the prospects of food waste management.

Lignocellulose integration to 1G-ethanol process using filamentous fungi: fermentation prospects of edible strain of Neurospora intermedia.

BACKGROUND: Integration of first- and second-generation ethanol processes is one among the alternate approaches that efficiently address the current socio-economic issues of the bioethanol sector. Edible filamentous fungus capable of utilizing pentoses from lignocelluloses and also possessing biomass application as potential animal feed component was used as the fermentation strain for the integration model. This study presents various fermentation aspects of using edible filamentous fungi in the integrated first and second generation ethanol process model. RESULTS: Fermentation of edible strain of N. intermedia on the integrated first and second-generation ethanol substrate (the mixture of dilute acid pretreated and enzymatically hydrolyzed wheat straw and thin stillage from the first-generation ethanol process), showed an ethanol yield maximum of 0.23 ± 0.05 g/g dry substrate. The growth of fungal pellets in presence of fermentation inhibitors (such as acetic acid, HMF and furfural) resulted in about 11 to 45% increase in ethanol production as compared to filamentous forms, at similar growth conditions in the liquid straw hydrolysate. Fungal cultivations in the airlift reactor showed strong correlation with media viscosity, reaching a maximum of 209.8 ± 3.7 cP and resulting in 18.2 ± 1.3 g/L biomass during the growth phase of fungal pellets. CONCLUSION: N. intermedia fermentation showed high sensitivity to the dilute acid lignocellulose pretreatment process, with improved fermentation performance at milder acidic concentrations. The rheological examinations showed media viscosity to be the most critical factor influencing the oxygen transfer rate during the N. intermedia fermentation process. Mycelial pellet morphology showed better fermentation efficiency and high tolerance towards fermentation inhibitors.

Effect of media rheology and bioreactor hydrodynamics on filamentous fungi fermentation of lignocellulosic and starch-based substrates under pseudoplastic flow conditions.

The aim of this work was to study how media rheology and bioreactor hydrodynamics would influence fermentation of lignocellulosic and starch-based substrates under pseudoplastic flow conditions. This was investigated using hydrolyzed wheat straw, wheat-based thin stillage and filamentous fungi as inoculum in bubble column, airlift and horizontal hybrid tubular/bubble column (textile bioreactor) bioreactors. The rheological models showed that the consistency index was dependent on biomass growth (R2 0.99) while the flow behavior index depended on biomass growth and suspended solid (R2 0.99). Oxygen transfer rate above 0.356 mmol-O2/L/h was needed for growing fungi with a cube-root growth rate constant of 0.03 g1/3/L1/3/h. At 1.4 VVM aeration the textile bioreactor performed better than others with minimal foaming, yields of 0.22 ±â€¯0.01 g/g and 0.47 ±â€¯0.01 g/g for ethanol and biomass, substrate consumption rate of 0.38 g/L/h. Operating the bioreactors with air-flowrate to cross-sectional area ratio of 8.75 × 10-3 (m3/s/m2) or more led to sustained foaming.

Does the second messenger cAMP have a more complex role in controlling filamentous fungal morphology and metabolite production?

The effect of second messenger cAMP on the physiological aspects of fungal cells such as pigmentation has been reported previously. However, their actual role in the cellular biochemical cascade that eventually affects the fungal growth morphology, such as mycelial pellet formation, is unclear. This article intends to open up the detailed study on the possible correlative effect of cAMP on the morphological and physiological growth aspects of filamentous fungi, with special emphasis on the industrial metabolite production.

Vegan-mycoprotein concentrate from pea-processing industry byproduct using edible filamentous fungi.

BACKGROUND: Currently around one billion people in the world do not have access to a diet which provides enough protein and energy. However, the production of one of the main sources of protein, animal meat, causes severe impacts on the environment. The present study investigates the production of a vegan-mycoprotein concentrate from pea-industry byproduct (PpB), using edible filamentous fungi, with potential application in human nutrition. Edible fungal strains of Ascomycota (Aspergillus oryzae, Fusarium venenatum, Monascus purpureus, Neurospora intermedia) and Zygomycota (Rhizopus oryzae) phyla were screened and selected for their protein production yield. RESULTS: A. oryzae had the best performance among the tested fungi, with a protein yield of 0.26 g per g of pea-processing byproduct from the bench scale airlift bioreactor cultivation. It is estimated that by integrating the novel fungal process at an existing pea-processing industry, about 680 kg of fungal biomass attributing to about 38% of extra protein could be produced for each 1 metric ton of pea-processing byproduct. This study is the first of its kind to demonstrate the potential of the pea-processing byproduct to be used by filamentous fungi to produce vegan-mycoprotein for human food applications. CONCLUSION: The pea-processing byproduct (PpB) was proved to be an efficient medium for the growth of filamentous fungi to produce a vegan-protein concentrate. Moreover, an industrial scenario for the production of vegan-mycoprotein concentrate for human nutrition is proposed as an integrated process to the existing PPI production facilities.

Integrated Process for Ethanol, Biogas, and Edible Filamentous Fungi-Based Animal Feed Production from Dilute Phosphoric Acid-Pretreated Wheat Straw.

Integration of wheat straw for a biorefinery-based energy generation process by producing ethanol and biogas together with the production of high-protein fungal biomass (suitable for feed application) was the main focus of the present study. An edible ascomycete fungal strain Neurospora intermedia was used for the ethanol fermentation and subsequent biomass production from dilute phosphoric acid (0.7 to 1.2% w/v) pretreated wheat straw. At optimum pretreatment conditions, an ethanol yield of 84 to 90% of the theoretical maximum, based on glucan content of substrate straw, was observed from fungal fermentation post the enzymatic hydrolysis process. The biogas production from the pretreated straw slurry showed an improved methane yield potential up to 162% increase, as compared to that of the untreated straw. Additional biogas production, using the syrup, a waste stream obtained post the ethanol fermentation, resulted in a combined total energy output of 15.8 MJ/kg wheat straw. Moreover, using thin stillage (a waste stream from the first-generation wheat-based ethanol process) as a co-substrate to the biogas process resulted in an additional increase by about 14 to 27% in the total energy output as compared to using only wheat straw-based substrates. ᅟ.

Mild-temperature dilute acid pretreatment for integration of first and second generation ethanol processes.

The use of hot-water (100°C) from the 1st generation ethanol plants for mild-temperature lignocellulose pretreatment can possibly cut down the operational (energy) cost of 2nd generation ethanol process, in an integrated model. Dilute-sulfuric and -phosphoric acid pretreatment at 100°C was carried out for wheat bran and whole-stillage fibers. Pretreatment time and acid type influenced the release of sugars from wheat bran, while acid-concentration was found significant for whole-stillage fibers. Pretreatment led up-to 300% improvement in the glucose yield compared to only-enzymatically treated substrates. The pretreated substrates were 191-344% and 115-300% richer in lignin and glucan, respectively. Fermentation using Neurospora intermedia, showed 81% and 91% ethanol yields from wheat bran and stillage-fibers, respectively. Sawdust proved to be a highly recalcitrant substrate for mild-temperature pretreatment with only 22% glucose yield. Both wheat bran and whole-stillage are potential substrates for pretreatment using waste heat from the 1st generation process for 2nd generation ethanol.

Valorization of sugar-to-ethanol process waste vinasse: A novel biorefinery approach using edible ascomycetes filamentous fungi.

The aim of the present work was to study the integration of edible ascomycetes filamentous fungi into the existing sugar- or molasses-to-ethanol processes, to grow on vinasse or stillage and produce ethanol and protein-rich fungal biomass. Two fungal strains, Neurospora intermedia and Aspergillus oryzae were examined in shake flasks and airlift-bioreactors, resulting in reduction of vinasse COD by 34% and viscosity by 21%. Utilization of glycerol and sugars were observed, yielding 202.4 or 222.8g dry fungal biomass of N. intermedia or A. oryzae respectively, per liter of vinasse. Integration of the current process at an existing ethanol facility producing about 100,000m3 of ethanol per year could produce around 200,000-250,000tons of dry fungal biomass (40-45% protein) together with about 8800-12,600m3 extra ethanol (8.8-12.6% of production-rate improvement).

Mycelial pellet formation by edible ascomycete filamentous fungi, Neurospora intermedia.

Pellet formation of filamentous fungi in submerged culture is an imperative topic of fermentation research. In this study, we report for the first time the growth of filamentous ascomycete fungus, Neurospora intermedia in its mycelial pellet form. In submerged culture, the growth morphology of the fungus was successfully manipulated into growing as pellets by modifying various cultivation conditions. Factors such as pH (2.0-10.0), agitation rate (100-150 rpm), carbon source (glucose, arabinose, sucrose, and galactose), the presence of additive agents (glycerol and calcium chloride) and trace metals were investigated for their effect on the pellet formation. Of the various factors screened, uniform pellets were formed only at pH range 3.0-4.0, signifying it as the most influential factor for N. intermedia pellet formation. The average pellet size ranged from 2.38 ± 0.12 to 2.86 ± 0.38 mm. The pellet formation remained unaffected by the inoculum type used and its size showed an inverse correlation with the agitation rate of the culture. Efficient glucose utilization was observed with fungal pellets, as opposed to the freely suspended mycelium, proving its viability for fast-fermentation processes. Scale up of the pelletization process was also carried out in bench-scale airlift and bubble column reactors (4.5 L).

Combined multidrug and Mycobacterium w vaccine therapy in patients with multibacillary leprosy.

Immunotherapy with Mycobacterium w vaccine was attempted in patients with borderline-borderline, borderline lepromatous (BL), or lepromatous leprosy (LL) to determine whether immunization can hasten recovery and reduce treatment time by invigorating cell-mediated immunity. Mycobacterium w, a nonpathogenic, rapidly growing, atypical mycobacterium, shares a number of common B and T cell determinants with Mycobacterium leprae and Mycobacterium tuberculosis. Patients receiving the vaccine had rapid clinical improvement and accelerated bacteriologic clearance. After treatment with vaccine for 2 years, 13 of 31 BL and LL patients were bacteriologically negative as were 5 of 25 controls. Vaccinated patients had one of two distinct histologic features, either an upgrading in the disease spectrum or complete clearance of granuloma. Some 80% of lepromin conversions were in BL and LL patients who received vaccine versus none and 14.3% of BL and LL controls, respectively. Thirteen of 17 vaccinated LL patients were released from treatment after 2 years in contrast to 2 of 15 controls.