Kinetic Study on the Effect of Substrate and Micronutrient Inhibition during Anaerobic Fermentation of Biohydrogen

There are several factors that influence the production of biohydrogen by dark fermentation including inoculum seeds, type and concentration of substrate, pH, temperature, presence of micronutrient and reactor configuration. Previous research has proven that the concentration of substrate and the presence of micronutrient will influence the yield and productivity of biohydrogen production. However, improvement of yield and productivity of the process can only be achieved once the system is under the optimum amount of substrate and micronutrient. Therefore, the best way to determine the effect of substrate concentration and presence of micronutrient is through kinetic study that was done using Monod model along with Andrews model. Besides that, the substrate inhibition effect also will be evaluated to determine the maximum substrate that needs to be supplied for maximum hydrogen production, and thus supplied the information for economic feasibility for fermentation process. In the meantime, the inhibition effect of adding the iron nanoparticles also had been evaluated in order to understand the interaction effect between iron nanoparticles and bacteria in term of catabolism reaction. It was found that increasing the substrate concentration more than 10 g/l will cause the inhibition to the system, in which it will slow down the reaction process and reduced the production of hydrogen. While the presence of iron NPs more than its optimum value (200 mg/l) will inhibit the bacterial growth and hence, affect the hydrogen production. For both cases, when the inhibition occurred at the respective concentration, it was found that the metabolic pathway was shifted to produce more hydrogen-consuming metabolite such as propionate acid, and thus, dropped the hydrogen production.

Mathematical Modelling of Bioethanol Production from Raw Sugar Beet Cossettes in a Horizontal Rotating Tubular Bioreactor

Alternative to the use of fossil fuels are biofuels (e.g., bioethanol, biodiesel and biogas), which are more environmentally friendly and which can be produced from different renewable resources. In this investigation, bioethanol production from raw sugar beet cossettes (semi-solid substrate) by yeast Saccharomyces cerevisiae in a horizontal rotating tubular bioreactor (HRTB) was studied. Obtained results show that HRTB rotation mode (constant or interval) and rotation speed have considerable impact on the efficiency of bioethanol production in the HRTB. The main goal of this research was to develop a non-structural mathematical model of bioethanol production from raw sugar beet cossettes in the HRTB. The established mathematical model of bioethanol production in the HRTB describes substrate utilization and product formation (glycerol, ethanol and acetate) and presumes negative impact of high substrate concentration on the working microorganism (substrate inhibition) by using Andrews inhibition kinetics. All simulations of bioethanol production in the HRTB were performed by using Berkeley Madonna software, version 8.3.14 (Berkeley Madonna, Berkeley, CA, USA). The established non-structural bioprocess model describes relatively well the bioethanol production from raw sugar beet cossettes in the HRTB.

Old Enzyme, New Role: The β-Glucosidase BglC of Streptomyces scabiei Interferes with the Plant Defense Mechanism by Hydrolyzing Scopolin

The beta-glucosidase BglC fulfills multiple functions in both primary metabolism and induction of pathogenicity of Streptomyces scabiei, the causative agent of common scab in root and tuber crops. Indeed, this enzyme hydrolyzes cellobiose and cellotriose to feed glycolysis with glucose directly and modifies the intracellular concentration of these cello-oligosaccharides, which are the virulence elicitors. The inactivation of bglC led to unexpected phenotypes such as the constitutive overproduction of thaxtomin A, the main virulence determinant of S. scabiei. In this work, we reveal a new target substrate of BglC, the phytoalexin scopolin. Removal of the glucose moiety of scopolin generates scopoletin, a potent inhibitor of thaxtomin A production. The hydrolysis of scopolin by BglC displayed substrate inhibition kinetics, which contrasts with the typical Michaelis–Menten saturation curve previously observed for the degradation of its natural substrate cellobiose. Our work, therefore, reveals that BglC targets both cello-oligosaccharide elicitors emanating from the hosts of S. scabiei, and the scopolin phytoalexin generated by the host defense mechanisms, thereby occupying a key position to fine-tune the production of the main virulence determinant thaxtomin A.

DEAD-box RNA Helicases Act as Nucleotide Exchange Factors for Casein Kinase 2

DDX RNA helicases promote RNA processing but DDX3X is also known to activate casein kinase 1 ϵ (CK1ϵ). Here we show that not only is protein kinase stimulation a latent property of other DDX proteins towards CK1ϵ, but that this extends to casein kinase 2 (CK2α2) as well. CK2α2 enzymatic activity is stimulated by a variety of DDX proteins and we identify DDX1/24/41/54 as physiological activators required for full kinase activity in vitro and in Xenopus embryos. Mutational analysis of DDX3X reveals that CK1 and CK2 kinase stimulation engages its RNA binding- but not catalytic motifs. Mathematical modelling of enzyme kinetics and stopped-flow spectroscopy converge that DDX proteins function as nucleotide exchange factor towards CK2α2 that reduce unproductive reaction intermediates and substrate inhibition. Our study reveals protein kinase stimulation by nucleotide exchange as a new principle in kinase regulation and an evolved function of DDX proteins.

The Stiffness of Cardiac Fibroblast Substrates Exerts a Regulatory Influence on Collagen Metabolism via α2β1 Integrin, FAK and Src Kinases

Information about mechanical strain in the extracellular space is conducted along collagen fibers connected with integrins and then transmitted within cells. An aim of the study is to verify the hypothesis that the stiffness of cardiac human fibroblast substrates exerts a regulatory effect on collagen metabolism via integrin α2β1 and downstream signaling. The experiments were performed on human cardiac fibroblasts cultured on stiff or soft polyacrylamide gels. Extracellular and intracellular collagen content, metalloproteinase-1 (MMP-1), metalloproteinase-9 (MMP-9) and expression of the α1 chain of the procollagen type I gene (Col1A1) were elevated in cultures settled on soft substrate. The substrate stiffness did not modify tissue inhibitors of matrix metalloproteinase capacity (TIMPs 1–4). Integrin α2β1 inhibition (TC-I 15) or α2 subunit silencing resulted in augmentation of collagen content within the culture. Expression of Col1A1 and Col3A1 genes was increased in TC-I 15-treated fibroblasts. Total and phosphorylated levels of both FAK and Src kinases were elevated in fibroblasts cultured on stiff substrate. Inhibition of FAK (FAK kinase inhibitor 14) or Src kinase (AZM 47527) increased collagen content within the culture. The substrate stiffness exerted a regulatory influence on collagen metabolism via integrin α2β1 and its downstream signaling (FAK and Src kinases) in cardiac fibroblasts.

Ascomycetes versus Spent Mushroom Substrate in Mycoremediation of Dredged Sediments Contaminated by Total Petroleum Hydrocarbons: The Involvement of the Bacterial Metabolism

Two mycoremediation approaches for the depletion of the total petroleum hydrocarbons in dredged sediments were compared: co-composting with spent mushroom substrate (SMS) from Pleurotus ostreatus and bioaugmentation with Lambertella sp. MUT 5852, an ascomycetes autochthonous to the sediment, capable of utilizing diesel oil its sole carbon source. After 28 days of incubation, 99% depletion was observed in presence of Lambertella sp. MUT 5852. No total petroleum hydrocarbon depletion was observed in sediment co-composting with the SMS after 60 days of incubation. 16S rDNA metabarcoding of the bacterial community was performed to evaluate the potential synergism between fungi and bacteria in the bioremediation process. The functional metagenomic prediction approach indicated that the biodiversity of the bacterial genera potentially involved in the degradation of TPH was higher in sediment bioaugmented with Lambertella sp. MUT 5852, which resulted in being mandatory for TPH depletion. Mechanisms of co-substrate inhibition of the hydrocarburoclastic bacterial species, due to the bioavailable organic matter of the SMS, are suggested to be involved in the observed kinetics of TPH depletion, failing in the case of SMS and successful in the case of Lambertella sp. MUT 5852.

Effect of Nickel as Stress Factor on Phenol Biodegradation by Stenotrophomonas maltophilia KB2

This study focuses on the phenol biodegradation kinetics by Stenotrophomonas maltophilia KB2 in a nickel-contaminated medium. Initial tests proved that a nickel concentration of 33.3 mg·L−1 caused a cessation of bacterial growth. The experiments were conducted in a batch bioreactor in several series: without nickel, at constant nickel concentration and at varying metal concentrations (1.67–13.33 g·m−3). For a constant Ni2+ concentration (1.67 or 3.33 g·m−3), a comparable bacterial growth rate was obtained regardless of the initial phenol concentration (50–300 g·m−3). The dependence µ = f (S0) at constant Ni2+ concentration was very well described by the Monod equations. The created varying nickel concentrations experimental database was used to estimate the parameters of selected mathematical models, and the analysis included different methods of determining metal inhibition constant KIM. Each model showed a very good fit with the experimental data (R2 values were higher than 0.9). The best agreement (R2 = 0.995) was achieved using a modified Andrews equation, which considers the metal influence and substrate inhibition. Therefore, kinetic equation parameters were estimated: µmax = 1.584 h−1, KS = 185.367 g·m−3, KIS = 106.137 g·m−3, KIM = 1.249 g·m−3 and n = 1.0706.

Contributions of UDP-Glucuronosyltransferases to Human Hepatic and Intestinal Metabolism of Ticagrelor and Inhibition of UGTs and Cytochrome P450 Enzymes by Ticagrelor and its Glucuronidated Metabolite

Ticagrelor is the first reversibly binding, direct-acting, oral P2Y12 receptor inhibitor. The contribution of UDP-glucuronosyltransferases (UGTs) enzymes to the metabolism of ticagrelor to its glucuronide conjugation, ticagrelor-O-glucuronide, in human liver microsomes (HLM) and human intestinal microsomes (HIM), was well characterized in the current study. The inhibition potential of human major UGTs by ticagrelor and ticagrelor-O-glucuronide was explored. The inhibitory effects of ticagrelor-O-glucuronide on cytochrome P450s (CYPs) enzymes were investigated as well. Ticagrelor glucuronidation exhibits substrate inhibition kinetics in both HLM and HIM with apparent Km values of 5.65 and 2.52 μM, Vmax values of 8.03 and 0.90 pmol min−1·mg protein−1, Ksi values of 1,343.0 and 292.9 respectively. The in vitro intrinsic clearances (Vmax/Km) for ticagrelor glucuronidation by HLM and HIM were 1.42 and 0.36 μl min−1·mg protein−1, respectively. Study with recombinant human UGTs suggested that multiple UGT isoforms including UGT1A9, UGT1A7, UGT1A3, UGT1A4, UGT1A1, UGT2B7 and UGT1A8 are involved in the conversion of ticagrelor to ticagrelor-O-glucuronide with UGT1A9 showing highest catalytic activity. The results were further supported by the inhibition studies on ticagrelor glucuronidation with typical UGT inhibitors in pooled HLM and HIM. Little or no inhibition of UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A9 and UGT2B7 by ticagrelor and ticagrelor-O-glucuronide was noted. Ticagrelor-O-glucuronide also exhibited limited inhibitory effects toward CYP2C8, CYP2D6 and CYP3A4. In contrast, ticagrelor-O-glucuronide weakly inhibited CYP2B6, CYP2C9 and CYP2C19 activity with apparent IC50 values of 45.0, 20.0 and 18.8 μM, respectively. The potential of ticagrelor-O-glucuronide to cause drug-drug interactions warrant further study.

Effect of feed slurry dilution and total solids on specific biogas production by anaerobic digestion in batch and semi-batch reactors

Biomass from various sources such as cow dung is a significant source of renewable energy (as biogas) in many regions globally, especially in India, Africa, Brazil, and China. However, biogas production from biomass such as cattle dung is a slow, inefficient biochemical process, and the specific biogas produced per kg of biomass is relatively small. The improvement of specific biogas production efficiency using various dilution ratios (and, hence, total solids [TS]) is investigated in this work. A wide range of feed dilution (FD) ratios of cow dung: water (CD: W) was tested in batch biogas digesters with total solids ranging from 1% to 12.5% and FD ratio ranging from 2:1 to 1:20. To further verify the results from the above batch experiments, semi-batch experiments representative of field-scale biodigesters were conducted. Semi-batch reactors have a steady-state process, unlike batch reactors, which have an unsteady state process. Our results suggested that specific biogas production (mL/g VS) increased continuously when the total solids decreased from 12.5% to 1% (or when dilution increased). Our experiments also indicate that the commonly used 1:1 feed dilution ratio (TS ~ 10% for cow dung) does not produce the maximum specific biogas production. The possible reason for this could be that anaerobic digestion at higher total solids is rate limited due to substrate inhibition, mass transfer limitations, and viscous mixing problems that arise at higher total solids concentration. Hence, a higher feed dilution ratio between 1:2 and 1:4 (TS between 4 and 6.7%) is recommended for a more efficient biomass utilization of cowdung. Empirical relationships were also developed for variation of specific biogas yield with the total solids content of the cow dung slurry. Graphic abstract