106 research outputs found
Selection of strain and optimization of mutanase production in submerged cultures of Trichoderma harzianum
Nineteen fungal strains belonging to different genera were tested for extracellular mutanase production in shaken flasks. The optimal enzymatic activity was achieved by Trichoderma harzianum F-470, a strain for which the mutanase productivity has not yet been published. Some of factors affecting the enzyme production in shaken flasks and aerated fermenter cultures have been standardized. Mandels mineral medium with initial pH 5.3, containing 0.25% mutan and inoculated with 10% of the 48-h mycelium, was the best for enzyme production. A slight mutanolytic activity was also found when sucrose, raffinose, lactose and melibiose were carbon sources. Application of optimized medium and cultural conditions, as well as use of a fermenter with automatic pH control set at pH 6.0 enabled to obtain a high mutanase yield (0.33 U/ml, 2.5 U/mg protein) in a short time (2-3 days). The enzyme in crude state was stable over a pH range of 4.5-6.0, and at temperatures up to 35 °C; its maximum activity was at 40 °C and at pH 5.5
Mutanase from Paenibacillus sp. MP-1 produced inductively by fungal α-1,3-glucan and its potential for the degradation of mutan and Streptococcus mutans biofilm
Laetiporus sulphureus is a source of α-1,3-glucan that can substitute for the commercially-unavailable streptococcal mutan used to induce microbial mutanases. The water-insoluble fraction of its fruiting bodies from 0.15 to 0.2% (w/v) induced mutanase activity in Paenibacillus sp. MP-1 at 0.35 μ ml−1. The mutanase extensively hydrolyzed streptococcal mutan, giving 23% of saccharification, and 83% of solubilization of glucan after 6 h. It also degraded α-1,3-polymers of biofilms, formed in vitro by Streptococcus mutans, even after only 3 min of contact
Measurements from the RV Ronald H. Brown and related platforms as part of the Atlantic Tradewind Ocean-Atmosphere Mesoscale Interaction Campaign (ATOMIC)
© The Author(s), 2021. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Quinn, P. K., Thompson, E. J., Coffman, D. J., Baidar, S., Bariteau, L., Bates, T. S., Bigorre, S., Brewer, A., de Boer, G., de Szoeke, S. P., Drushka, K., Foltz, G. R., Intrieri, J., Iyer, S., Fairall, C. W., Gaston, C. J., Jansen, F., Johnson, J. E., Krueger, O. O., Marchbanks, R. D., Moran, K. P., Noone, D., Pezoa, S., Pincus, R., Plueddemann, A. J., Poehlker, M. L., Poeschl, U., Melendez, E. Q., Royer, H. M., Szczodrak, M., Thomson, J., Upchurch, L. M., Zhang, C., Zhang, D., & Zuidema, P. Measurements from the RV Ronald H. Brown and related platforms as part of the Atlantic Tradewind Ocean-Atmosphere Mesoscale Interaction Campaign (ATOMIC). Earth System Science Data, 13(4), (2021): 1759-1790, https://doi.org/10.5194/essd-13-1759-2021.The Atlantic Tradewind Ocean-Atmosphere Mesoscale Interaction Campaign (ATOMIC) took place from 7 January to 11 July 2020 in the tropical North Atlantic between the eastern edge of Barbados and 51∘ W, the longitude of the Northwest Tropical Atlantic Station (NTAS) mooring. Measurements were made to gather information on shallow atmospheric convection, the effects of aerosols and clouds on the ocean surface energy budget, and mesoscale oceanic processes. Multiple platforms were deployed during ATOMIC including the NOAA RV Ronald H. Brown (RHB) (7 January to 13 February) and WP-3D Orion (P-3) aircraft (17 January to 10 February), the University of Colorado's Robust Autonomous Aerial Vehicle-Endurant Nimble (RAAVEN) uncrewed aerial system (UAS) (24 January to 15 February), NOAA- and NASA-sponsored Saildrones (12 January to 11 July), and Surface Velocity Program Salinity (SVPS) surface ocean drifters (23 January to 29 April). The RV Ronald H. Brown conducted in situ and remote sensing measurements of oceanic and atmospheric properties with an emphasis on mesoscale oceanic–atmospheric coupling and aerosol–cloud interactions. In addition, the ship served as a launching pad for Wave Gliders, Surface Wave Instrument Floats with Tracking (SWIFTs), and radiosondes. Details of measurements made from the RV Ronald H. Brown, ship-deployed assets, and other platforms closely coordinated with the ship during ATOMIC are provided here. These platforms include Saildrone 1064 and the RAAVEN UAS as well as the Barbados Cloud Observatory (BCO) and Barbados Atmospheric Chemistry Observatory (BACO). Inter-platform comparisons are presented to assess consistency in the data sets. Data sets from the RV Ronald H. Brown and deployed assets have been quality controlled and are publicly available at NOAA's National Centers for Environmental Information (NCEI) data archive (https://www.ncei.noaa.gov/archive/accession/ATOMIC-2020, last access: 2 April 2021). Point-of-contact information and links to individual data sets with digital object identifiers (DOIs) are provided herein.NOAA's Climate Variability and Predictability Program provided funding under NOAA CVP NA19OAR4310379, GC19-301, and GC19-305. The Joint Institute for the Study of the Atmosphere and Ocean (JISAO) supported this study under NOAA cooperative agreement NA15OAR4320063. Additional support was provided by the NOAA's Uncrewed Aircraft Systems (UAS) Program Office, NOAA's Physical Sciences Laboratory, and NOAA AOML's Physical Oceanography Division. The NTAS project is funded by the NOAA's Global Ocean Monitoring and Observing Program (CPO FundRef number 100007298), through the Cooperative Institute for the North Atlantic Region (CINAR) under cooperative agreement NA14OAR4320158
Fermentation of deproteinized cheese whey powder solutions to ethanol by engineered Saccharomyces cerevisiae : effect of supplementation with corn steep liquor and repeated-batch operation with biomass recycling by flocculation
The lactose in cheese whey is an interesting
substrate for the production of bulk commodities such as
bio-ethanol, due to the large amounts of whey surplus
generated globally. In this work, we studied the performance
of a recombinant Saccharomyces cerevisiae strain
expressing the lactose permease and intracellular ß-galactosidase
from Kluyveromyces lactis in fermentations of
deproteinized concentrated cheese whey powder solutions.
Supplementation with 10 g/l of corn steep liquor significantly
enhanced whey fermentation, resulting in the production
of 7.4% (v/v) ethanol from 150 g/l initial lactose in
shake-flask fermentations, with a corresponding productivity
of 1.2 g/l/h. The flocculation capacity of the yeast
strain enabled stable operation of a repeated-batch process
in a 5.5-l air-lift bioreactor, with simple biomass recycling
by sedimentation of the yeast flocs. During five consecutive
batches, the average ethanol productivity was 0.65 g/l/h
and ethanol accumulated up to 8% (v/v) with lactose-toethanol
conversion yields over 80% of theoretical. Yeast
viability (>97%) and plasmid retention (>84%) remained
high throughout the operation, demonstrating the stability
and robustness of the strain. In addition, the easy and
inexpensive recycle of the yeast biomass for repeated utilization
makes this process economically attractive for
industrial implementation.Fundação para a Ciência e a Tecnologia (FCT)LACTOGAL-Produtos Alimentares S.A.Companhia Portuguesa de Amidos, S.A
Arp2/3 complex interactions and actin network turnover in lamellipodia
Cell migration is initiated by lamellipodia—membrane-enclosed sheets of cytoplasm containing densely packed actin filament networks. Although the molecular details of network turnover remain obscure, recent work points towards key roles in filament nucleation for Arp2/3 complex and its activator WAVE complex. Here, we combine fluorescence recovery after photobleaching (FRAP) of different lamellipodial components with a new method of data analysis to shed light on the dynamics of actin assembly/disassembly. We show that Arp2/3 complex is incorporated into the network exclusively at the lamellipodium tip, like actin, at sites coincident with WAVE complex accumulation. Capping protein likewise showed a turnover similar to actin and Arp2/3 complex, but was confined to the tip. In contrast, cortactin—another prominent Arp2/3 complex regulator—and ADF/cofilin—previously implicated in driving both filament nucleation and disassembly—were rapidly exchanged throughout the lamellipodium. These results suggest that Arp2/3- and WAVE complex-driven actin filament nucleation at the lamellipodium tip is uncoupled from the activities of both cortactin and cofilin. Network turnover is additionally regulated by the spatially segregated activities of capping protein at the tip and cofilin throughout the mesh
Enzymatic hydrolysis of sorghum straw using native cellulase produced by T. reesei NCIM 992 under solid state fermentation using rice straw
Cellulose is a major constituent of renewable lignocellulosic waste available in large quantities and is considered the most important reservoir of carbon for the production of glucose, for alternative fuel and as a chemical feedstock. Over the past decade, the emphasis has been on the enzymatic hydrolysis of cellulose to glucose and the efficiency of which depends on source of cellulosic substrate, its composition, structure, pretreatment process, and reactor design. In the present study, efforts were made to produce cellulase enzyme using rice straw. The produced enzyme was used for the hydrolysis of selected lignocellulosic substrate, i.e., sorghum straw. When rice straw was used as a substrate for cellulase production under solid state fermentation, the highest enzyme activity obtained was 30.7 FPU/gds, using T. reesei NCIM 992. 25 FPU/g of cellulase was added to differently treated (native, alkali treated, alkali treated followed by 3% acid treated and alkali treated followed by 3 and 5% acid treated) sorghum straw and hydrolysis was carried out at 50 °C for 60 h. 42.5% hydrolysis was obtained after 36 h of incubation. Optimization of enzyme loading, substrate concentration, temperature, time and buffer yielded a maximum of 546.00 ± 0.55 mg/g sugars (54.60 ± 0.44 g/l) with an improved hydrolysis efficiency of 70 ± 0.45%. The enzymatic hydrolyzate can be used for fermentation of ethanol by yeasts
Microtubules as Platforms for Assaying Actin Polymerization In Vivo
The actin cytoskeleton is continuously remodeled through cycles of actin filament assembly and disassembly. Filaments are born through nucleation and shaped into supramolecular structures with various essential functions. These range from contractile and protrusive assemblies in muscle and non-muscle cells to actin filament comets propelling vesicles or pathogens through the cytosol. Although nucleation has been extensively studied using purified proteins in vitro, dissection of the process in cells is complicated by the abundance and molecular complexity of actin filament arrays. We here describe the ectopic nucleation of actin filaments on the surface of microtubules, free of endogenous actin and interfering membrane or lipid. All major mechanisms of actin filament nucleation were recapitulated, including filament assembly induced by Arp2/3 complex, formin and Spir. This novel approach allows systematic dissection of actin nucleation in the cytosol of live cells, its genetic re-engineering as well as screening for new modifiers of the process
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