Increasing Substrate Concentration Research Articles

Investigating the efficiency of a two-stage anaerobic-aerobic process for the treatment of confectionery industry wastewaters with simultaneous production of biohydrogen and polyhydroxyalkanoates.

The scope of the current study was to investigate the efficiency of a two-stage anaerobic-aerobic process for the simultaneous treatment and valorization of selective wastewater streams from a confectionary industry. The specific wastewater (confectionary industry wastewater, CIW) was a mixture of the rinsing eluting during washing of the cauldrons in which jellies and syrups were produced, and contained mainly readily fermentable sugars, being thus of high organic load. The first stage of the process was the dark fermentation (DF) of the CIW in continuous, attached-biomass systems, in which the effect on hydrogen yields and distribution of metabolites were studied for different packing materials (ceramic or plastic), hydraulic retention times, HRTs (12 h–30 h) and feed substrate concentration (20 g COD/L- 50 g COD/L). In the second stage, the effectiveness of the aerobic treatment of the DF effluents was evaluated in terms of the reduction of the organic load and the production of polyhydroxyalkanoates (PHAs) through an enriched mixed microbial culture (MMC). The MMC was developed in a continuous draw and fill system, in which the accumulation potential of PHAs was studied. It was shown that the hydrogen production rates decreased for increasing substrate concentration and HRTs, with a maximum of 12.70 ± 0.35 m3 H2/m3 initial CIW achieved for the lowest HRT and feed concentration and using ceramic beads as packing material. Butyrate, acetate and lactate were the main metabolites generated in all cases, in different ratios. The distribution of metabolites during DF was shown to highly affect the efficiency of the second process in terms of both the reduction of organic load and the PHAs yields. The highest removal of organic load achieved after 48 h of aerobic treatment was 84.0 ± 0.9 %, whereas the maximum PHAs yield was 21.46 ± 0.13 kg PHAs/m3 initial CIW.

Xylitol fermentation characteristics with a newly isolated yeast Wickerhamomyces anomalus WA

Xylitol is an increasingly popular functional food additive, and the newly isolated yeast Wickerhamomyces anomalus WA has shown extensive substrate utilization capability, with the ability to grow on hexose (d-galactose, d-glucose, d-mannose, l-fructose, and d-sorbose) and pentose (d-xylose and l-arabinose) substrates, as well as high tolerance to xylose at concentrations of up to 300 g/L. Optimal xylitol fermentation conditions were achieved at 32 °C, 140 rpm, pH 5.0, and initial cell concentration OD600 of 2.0, with YP (yeast extract 10 g/L, peptone 20 g/L) as the optimal nitrogen source. Xylitol yield increased from 0.61 g/g to 0.91 g/g with an increase in initial substrate concentration from 20 g/L to 180 g/L. Additionally, 20 g/L glycerol was found to be the optimal co-substrate for xylitol fermentation, resulting in an increase in xylitol yield from 0.82 g/g to 0.94 g/g at 140 rpm, enabling complete conversion of xylose to xylitol.

Interactive effects of microbial functional diversity and carbon availability on decomposition – A theoretical exploration

Microbial functional diversity in litter and soil has been hypothesized to affect the rate of decomposition of organic matter and other soil ecosystem functions. However, there are no clear theoretical expectations on how these effects might change with substrate availability, heterogeneity in the substrate chemistry, and different aspects of functional diversity itself (number of microbial groups vs. distribution of functional traits). To explore how these factors shape the decomposition-diversity relation, we carry out numerical experiments using a flexible reaction network comprising microbial processes and interactions with bioavailable carbon (extracellular degradation, uptake, respiration, growth, and mortality), and ecological processes (competition among the different groups). We also considered diverse carbon substrates, in terms of varying nominal oxidation state of carbon (NOSC). The reaction network was used to test the effects of (i) number of microbial groups, (ii) number of carbon pools, (iii) microbial functional diversity, and (iv) amount of bioavailable carbon. We found that the decomposition rate constant increases with increasing substrate concentration and heterogeneity, as well as with increasing microbial functional diversity or variance of microbial traits, albeit these biological factors are less important. The multivariate dependence of the decomposition rate constant (and other decomposition and microbial growth metrics) on substrate and microbial factors can be described using power laws with exponents lower than one, indicating that diversity effects on decomposition and microbial growth are reduced at high substrate concentration and heterogeneity, or at high microbial diversity.

Human soluble CD39 displays substrate inhibition in a substrate-specific manner

CD39 (ectonucleoside triphosphate diphosphohydrolase-1; ENTPD1) metabolizes extracellular ATP and ADP to AMP. AMP is subsequently metabolized by CD79 to adenosine. CD39 activity is therefore a key regulator of purinergic signalling in cancer, thrombosis, and autoimmune diseases. In this study we demonstrate that soluble, recombinant CD39 shows substrate inhibition with ADP or ATP as the substrate. Although CD39 activity initially increased with increasing substrate concentration, at high concentrations of ATP or ADP, CD39 activity was markedly reduced. Although the reaction product, AMP, inhibits CD39 activity, insufficient AMP was generated under our conditions to account for the substrate inhibition seen. In contrast, inhibition was not seen with UDP or UTP as substrates. 2-methylthio-ADP also showed no substrate inhibition, indicating the nucleotide base is an important determinant of substrate inhibition. Molecular dynamics simulations revealed that ADP can undergo conformational rearrangements within the CD39 active site that were not seen with UDP or 2-methylthio-ADP. Appreciating the existence of substrate inhibition of CD39 will help the interpretation of studies of CD39 activity, including investigations into drugs that modulate CD39 activity.

Assessment of Codispensing Patterns of Mirabegron and Prespecified CYP2D6 Substrates in Patients with Overactive Bladder.

Patients with overactive bladder (OAB) experience sudden, intense urges to urinate, which may include urge urinary incontinence and nocturia. Pharmacotherapy includes β3-adrenergic receptor agonists such as mirabegron; however, mirabegron contains a label warning for cytochrome P450 (CYP) 2D6 inhibition, making coadministration with CYP2D6 substrates require monitoring and dose adjustment to avoid unintended increases in substrate concentration. To understand the codispensing patterns of mirabegron among patients using ten predefined CYP2D6 substrates with and before mirabegron dispensing. This retrospective claims database analysis used the IQVIA PharMetrics® Plus Database to assess codispensing of mirabegron with ten predefined CYP2D6 substrate groups identified on the basis of medications most frequently prescribed in the UnitedStates, those with high susceptibility to CYP2D6 inhibition, and those with evidence for exposure-related toxicity. Patients had to be ≥18 years old before initiation of the CYP2D6 substrate episode that overlapped with mirabegron. The cohort entry period was November 2012 to September 2019, and the overall study period was 1 January 2011 to 30 September 2019. Comparisons of patient profiles at dispensing were made between time periods with and before mirabegron use in the same patient. Descriptive statistics were used to assess the number of exposure episodes, total duration of exposure, and median duration of exposure of CYP2D6 substrate dispensing with and before mirabegron. CYP2D6 substrate exposure periods totaling ≥9000 person-months were available before overlapping exposure to mirabegron for all ten CYP2D6 substrate cohorts. Median codispensing duration for chronically administered CYP2D6 substrates was 62 (interquartile range [IQR] 91) days for citalopram/escitalopram, 71 (105) days for duloxetine/venlafaxine, and 75 (115) days for metoprolol/carvedilol; median codispensing duration for acutely administered CYP2D6 substrates was 15 (33) days for tramadol and 9 (18) days for hydrocodone. In this claims database analysis, the dispensing patterns of CYP2D6 substrates with mirabegron displayed frequent overlapping of exposure. Thus, a need exists to better understand the outcomes experienced by patients with OAB who are at increased risk for drug‒drug interactions when taking multiple CYP2D6 substrates concurrently with a CYP2D6 inhibitor.

Leveraging the concretes calcification with carbonic anhydrase produced by Alcaligenes faecalis GA(B) (Mn847724.1)

Concrete is an integrated substance that is composed of cement, water, and aggregates. The aim of this study was to biocalcify concretes with bacterial carbonic anhydrase (CA). To achieve this, a soil sample taken from a construction site in Minna Nigeria, was serially diluted and plated on nutrient agar with CaCO3 added. After 7, 14 and 28 days of curing by water immersion, the compressive strength of the concrete was determined and the effects of CA on the concretes were investigated. Alcaligenes faecalis was identified as the isolate with the better ability for CA production. The CA has a mean activity of 0.0351 U/ml, an increase in substrate concentration on CA activity from 1 to 7 mM, an optimum temperature of 50 °C and pH of 8.5. Concretes calcified with CA from A. faecalis gained a strength of 48.29 % after 28 days of curing by water immersion. The results demonstrated that CA has an enormous potential in concrete biocalcification.

Biohydrogen production: A new controllability criterion for analyzing the impacts of dark fermentation conditions

Biohydrogen production from renewable resources using dark fermentation has become an increasingly attractive solution in sustainable global energy supply. So far, there has been no report on the controllability analysis of biohydrogen production using dark fermentation. Process controllability is a crucial factor determining process feasibility. This paper presents a new criterion for assessing biohydrogen process controllability based on PI control. It proposes the critical loop gain derived via Routh stability analysis as a measure of process controllability. Results show that the dark fermentation using the bacteria from anaerobic dairy sludge and substrate source from sugarcane vinasse can lead to a highly controllable process with a critical loop gain value of 4.3. For the two other cases, an increase of substrate concentration from 10 g/L to 40 g/L substantially reduces the controllability. The proposed controllability criterion is easily adopted to assess the process feasibilty based on experimental data.

Influence of Degree of Polymerization of Low-Molecular-Weight Chitosan Oligosaccharides on the α-Glucosidase Inhibition.

Chitosan oligosaccharide (COS) is a bioactive compound derived from marine by-products. COS consumption has been demonstrated to lower the risk of diabetes. However, there are limited data on the inhibitory effect of low-molecular-weight COSs with different degrees of polymerization (DP) on α-glucosidase. This study investigates the α-glucosidase inhibitory activity of two low-molecular-weight COSs, i.e., S-TU-COS with DP2−4 and L-TU-COS with DP2−5, both of which have different molecular weight distributions. The inhibition constants of the inhibitors binding to free enzymes (Ki) and an enzyme−substrate complex (Kii) were investigated to elucidate the inhibitory mechanism of COSs with different chain lengths. The kinetic inhibition model of S-TU-COS showed non-completive inhibition results which are close to the uncompetitive inhibition results with Ki and Kii values of 3.34 mM and 2.94 mM, respectively. In contrast, L-TU-COS showed uncompetitive inhibition with a Kii value of 5.84 mM. With this behavior, the IC50 values of S-TU-COS and L-TU-COS decreased from 12.54 to 11.84 mM and 20.42 to 17.75 mM, respectively, with an increasing substrate concentration from 0.075 to 0.3 mM. This suggests that S-TU-COS is a more potent inhibitor, and the different DP of COS may cause significantly different inhibition (p < 0.05) on the α-glucosidase activity. This research may provide new insights into the production of a COS with a suitable profile for antidiabetic activity.

Degradation kinetics of aromatic VOCs polluted wastewater by functional bacteria at laboratory scale

Reaction kinetics in biodegradation process is the basis and key of bioremediation technology, which can be used to predict the interaction between microorganisms and environmental states in the system. In this study, the kinetic model (Monod, Moser, Tessier and Cotonis) and kinetic parameters of aerobic biodegradation of functional bacteria in simulated wastewater polluted by aromatic volatile organic compounds (VOCs) were determined by shaking flask experiment. Monod, Moser, Tessier and Contois models were used to fit the experimental data and determine the kinetic parameters based on nonlinear regression analysis. Experimental results demonstrated that the removal rate of aromatic VOCs at 72 h was between 34.78 and 99.75% depending on the initial concentration of aromatic VOCs. The specific growth rate μ and degradation rate q increased with the increase of substrate concentration. The model of Monod, Moser and Tessier could be used to simulate microbial growth and substrate degradation in simulated aromatic VOCs polluted wastewater. Then the model and corresponding kinetic parameters were used to predict the limit concentration of biodegradation and provide theoretical support for the subsequent dynamic simulation and field engineering.

SmiA is a hybrid priming/scaffolding adaptor for the LonA protease in Bacillus subtilis

Regulatory proteolysis targets properly folded clients via a combination of cis-encoded degron sequences and trans-expressed specificity factors called adaptors. SmiA of Bacillus subtilis was identified as the first adaptor protein for the Lon family of proteases, but the mechanism of SmiA-dependent proteolysis is unknown. Here, we develop a fluorescence-based assay to measure the kinetics of SmiA-dependent degradation of its client SwrA and show that SmiA–SwrA interaction and the SwrA degron were both necessary, but not sufficient, for proteolysis. Consistent with a scaffolding adaptor mechanism, we found that stoichiometric excess of SmiA caused substrate-independent inhibition of LonA-dependent turnover. Furthermore, SmiA was strictly required even when SwrA levels were high suggesting that a local increase in substrate concentration mediated by the scaffold was not sufficient for proteolysis. Moreover, SmiA function could not be substituted by thermal denaturation of the substrate, consistent with a priming adaptor mechanism. Taken together, we conclude that SmiA functions via a mechanism that is a hybrid between scaffolding and priming models.

Cu/nucleotide coordination self-assembling to in situ regenerate NAD(P)+ and co-immobilize dehydrogenase with robust activity and stability

Enzyme is an efficient green catalyst, but its vulnerability and difficulties of separation from products limited the application in industry. Furthermore, a large number of oxidase enzymes rely on dihydronicotinamide adenine dinucleotide (NADH) or dihydronicotinamide adenine dinucleotide phosphate (NADPH) cofactors. These cofactors are too expensive to be used in industrial processes. Thus, cofactor regeneration is an important method to avoid the consumption of large quantities of oxidized cofactor NAD(P)+ in enzyme-catalyzed reactions. In this paper, Cu based metal nucleotides coordination polymers (MNCPs) with NAD(P)H oxidase activity were constructed by self-assembly of copper ions and nucleotides. Cu2+/AMP and Cu2+/GMP showed high catalytic activity and high reusability. Then, Cu2+/AMP with NADH oxidase activity was combined with NAD+ dependent alcohol dehydrogenase (ADH) to construct a bienzyme cascade system with circulating cofactor, which applied to the conversion of benzyl alcohol to benzaldehyde. In this system, ADH exhibited high activity due to the increase of substrate concentration and the inhibition of the diffusion of intermediate products. Besides, the bienzyme cascade system also showed high stability and reusability. After 8 cycles, the immobilized ADH remained more than 57% of the initial activity. The prepared bienzyme cascade system can extensively extended its application in industrial production.

The prevalence of Fabry disease among 1009 unrelated patients with hypertrophic cardiomyopathy: a Russian nationwide screening program using NGS technology

BackgroundThere is a vast number of screening studies described in the literature from the beginning of the twenty-first century to the present day. Many of these studies are related to the estimation of Fabry disease (FD) morbidity among patients from high-risk groups, including adult patients with hypertrophic cardiomyopathy (HCM) and left ventricular hypertrophy (LVH). These studies show diverse detection frequencies (0–12%) depending on the methodology. Our study is the only example of large-scale selective FD screening based on the implementation of next-generation sequencing technology (NGS) as a first-level test to estimate FD morbidity in the Russian population over 18 years of age burdened with HCM.MethodsThe study included 1009 patients (578 males and 431 females), with a median age of 50 years, who were diagnosed with HCM according to current clinical guidelines. In the first stage of screening, all patients underwent molecular genetic testing (NGS method) of target regions. These regions included the coding sequences of 17 genes and mutations that can lead to the development of HCM. Lysosomal globotriaosylsphingosine (lyso-Gb3) concentrations and α-galactosidase A (α-gal A) enzyme activity were measured in the second stage of screening to reveal pathogenic or likely pathogenic variants in the GLA gene.ResultsWe revealed 8 (0.8%) patients (3 (37.5%) males and 5 (62.5%) females) with an average age of 59 ± 13.3 years who had pathogenic, likely pathogenic variants and variants of uncertain significance (VUS) in the GLA gene (NM_000169.2) as a result of selective screening of 1009 Russian patients with HCM. FD was confirmed via biochemical tests in a male with the pathogenic variant c.902G > A, p.R301Q as well as in two females with likely pathogenic variants c.897C > A, p.D299E and c.1287_1288dup, p.*430Fext*?. These tests showed reduced enzymatic activity and increased substrate concentration. However, a female with the pathogenic variant c.416A > G, p.N139S and with normal enzymatic activity only had increased substrate concentrations. The revealed nucleotide variants and high values of biochemical indicators (lyso-Gb3) in these 4 patients allowed us to estimate the FD diagnosis among 1009 Russian patients with HCM. Mild extracardiac manifestations were observed in these four patients; however, both biochemical values within the reference range in females with the c.971T > G, p.L324W (VUS) variant. α-gal A activity and lyso-Gb3 concentrations were also within the normal range in two males with hemizygous variants, c.546T > C, p.D182D and c.640-794_640-791del (we regarded them as VUS), and in one female with the c.427G > A, p.A143T variant (with conflicting interpretations of pathogenicity).ConclusionThe prevalence rate of FD among 1,009 adult Russian patients with HCM was 0.4%. We recommend FD screening among adult patients of both sexes with HCM and an undefined genetic cause via NGS method with subsequent analysis of α-gal A activity and lyso-Gb3 concentration in patients with pathogenic, likely pathogenic variants, and VUS. This strategy identifies patients with an atypical form of FD that is characterized by high residual activity of α-gal A, low concentrations of lyso-Gb3, and minor extracardiac manifestations.

Rapid cultivation of anammox sludge based on Ca-alginate cell beads.

Current gel entrapment technology has certain advantages for the enrichment of anammox sludge. In this study, the optimal preparation conditions and cultivation equipment of Ca-alginate cell beads for the culturing anammox sludge were proposed. The preparation parameters of the Ca-alginate cell beads were as follows: 3% sodium alginate, 4% CaCl2, VSA:Vcell = 1:1, a drop height of 9 cm, stirring speed of 300 rpm, and cross-linking time of 24 h. The prepared cell beads were regular spheres with a uniform size and hard texture. Throughout the 9 days of cultivation, the number of anammox bacteria in the Ca-alginate cell beads was 4.3 times that of the initial sludge, and the color of the cell beads changed from yellowish-brown to reddish-brown. Scanning electron microscopy (SEM) analysis showed that the SA gel beads had a good microporous structure. The fluorescence in situ hybridization (FISH) results illustrated that the bacteria were mostly dispersed inside the Ca-alginate cell beads. Additionally, the qPCR results implied that only a relatively small amount of anammox biomass (2.74×106 copies/gel-bead) was required to quickly start the anammox process. The anammox bacteria in the Ca-alginate cell beads grew with a fast growth rate in a short period and exhibited high activity due to diffusion limitations. In addition, the anammox bacteria cultivated in the Ca-alginate cell beads could adapt to the increase in substrate concentration in a short period. The optimal incubation time of this gel entrapment method for anammox sludge was no more than 17 days under the experimental conditions of this work. Therefore, this simple and practicable gel entrapment method may serve as a suitable pre-culture means for the rapid enrichment of anammox bacteria.

Study Enhanced Enzyme Diffusion with High-Speed Single Nanoparticle Rotational and Translational Tracking

Catalytic enzymes exhibiting enhanced motion have drawn extensive attention over the past decade; nevertheless, little is known about the effect on the environment induced by enzymes. Herein, we studied the active urease system by simultaneously monitoring the diffusion of single anisotropic gold nanorods (AuNRs) with high speed dark-field imaging. We found both the translational and the rotational diffusion coefficients of AuNRs were enhanced but with inconsistent degrees, indicating the catalytic reaction had a minor effect on the physiochemical properties of the environment according to the Stokes-Einstein equation. With the increase of substrate concentration, the diffusion of AuNRs showed increased spatial but decreased temporal heterogeneity. Additionally, high speed imaging revealed AuNRs could experience intermittent ballistic motion for tens of milliseconds. These results imply inhomogeneous distribution of enzymes in free solution induced by active enzymatic reactions.

Effects of substrate concentration and glycine betaine on nitrogen removal and microbial communities in the sulfide autotrophic denitrification and Anammox coupling system

The effects of substance concentration and glycine betaine (GB) on nitrogen removal and microbial communities in the sulfide autotrophic denitrification (SAD) and Anammox coupling process were investigated. When the ratio of NH4+-N/NO3−-N/S2− was 1:1.26:0.79, the ammonia removal efficiency (ARE) was 96.8%, and total nitrogen removal efficiency (TNRE) was about 94.8%. When the substance concentration increased gradually, ARE and TNRE decreased to 53.7% and 71.2%, respectively. However, when 0.5 mM GB was added, ARE and TNRE improved to 62.1% and 77.6%, respectively. Batch tests also showed that when 0.5 mM GB was added, the removal efficiency increased mostly. High-throughput sequencing illustrated that the Anammox bacteria abundance decreased, while the abundance of Thiobacillus increased with increasing substrate concentration. The abundance of Candidatus Kuenenia and Candidatus Brocadia in the upper of UASB reactor was significantly higher than that in the bottom. The main functional bacteria in the bottom of UASB reactor was Thiobacillus.

Modeling of biohydrogen production using generalized multi-scale kinetic model: Impacts of fermentation conditions

This paper presents a new multi-scale kinetic model built upon the multi-stage growth Hypothesis for predicting biohydrogen production. The proposed model represents the significant factors affecting biohydrogen production using a sum of first-order kinetic terms with varying dynamics from slow to fast one. The current work investigates 52 case studies of biohydrogen production that show the double first-order kinetic model provides the best modeling fitness (R2 > 0.99). This result suggests two prevalent pathways or microbial groups with distinct dynamics (i.e., fast and slow modes) in biohydrogen production. An increase in temperature (30 °C–43 °C) or substrate concentration (10 g/L to 40 g/L) and the use of simple substrates or mixed cultures can increase the fast-mode dominance up to 100% contribution. Model analysis suggests that the fast mode corresponds to the butyrate production pathway, the growth-associated hydrogen-producing activity, the easily-biodegradable substrates, or the quick hydrogen-producing groups.

Combined Probe Strategy to Increase the Enzymatic Digestion Rate and Accelerate the Renal Radioactivity Clearance of Peptide Radiotracers.

High and sustained renal radioactivity accumulation is a major challenge in peptide-based radionuclide imaging and therapy. However, neutral endopeptidase (NEP)-based enzymatic hydrolysis to release and excrete the radioactive fragments has been proven to be an effective and promising way to reduce renal accumulation. Despite the improvement, the effect is still far from being satisfactory. To further reduce kidney uptake, we studied the relationship between the enzymatic reaction rate and the substrate concentration and came up with a combined probe strategy. Model compounds Boc-MVK-Dde and Boc-MFK-Dde were used for an in vitro enzymatic digestion study. NOTA-Exendin 4 and NOTA-MVK-Exendin 4 were labeled with 64Cu for in vivo dose-dependent micro-positron emission tomography (PET) studies. Groups 1 and 2 were injected with 0.2 and 0.8 nmol of 64Cu-NOTA-Exendin 4, respectively. Groups 3-6 were injected with 0.2, 0.8, 1.0, and 1.4 nmol of 64Cu-NOTA-MVK-Exendin 4, respectively. Groups 7 and 8 were co-injected with 0.2 nmol of 64Cu-NOTA-MVK-Exendin 4 and NOTA-MVK-PEG5K (1.3 and 2.6 nmol). The radioactivity uptakes were determined and compared within and among the groups. The in vitro cleavage study for both Boc-MVK-Dde and Boc-MFK-Dde indicated that within a certain concentration range, the enzyme digestion rate increased with increasing substrate concentration. The microPET images showed that the renal clearance could be accelerated significantly by increasing the injection dose of 64Cu-NOTA-MVK-Exendin 4, with the kidney uptakes being 60.98, 43.01, and 16.10 % ID/g at 1 h for groups 3, 4 and 5, respectively. Unfortunately, the tumor uptakes were also significantly inhibited as the injected dose of the tracer increased. However, with the co-injection of NOTA-MVK-PEG5K, the renal accumulation was significantly decreased without hampering the tumor uptake. As a result, the tumor-to-kidney ratios were significantly improved, which were 1.93, 3.47, 1.74, and 3.38 times that of group 3 at 1, 4, 24, and 48 h, respectively. The enzymatic reaction rate of NEP is dependent on the concentration of the substrates both in vitro and in vivo. The combined probe strategy developed in this study can dramatically reduce the renal accumulation of a peptide radioligand without affecting the tumor uptake, which shows great potential in peptide-based radiotheranostics.

Controls on turnover of marine dissolved organic matter—testing the null hypothesis of purely concentration‐driven uptake: Comment on Shen and Benner, “Molecular properties are a primary control on the microbial utilization of dissolved organic matter in the ocean”

Controls on turnover of marine dissolved organic matter—testing the null hypothesis of purely concentration‐driven uptake: Comment on Shen and Benner, “Molecular properties are a primary control on the microbial utilization of dissolved organic matter in the ocean”

A practical approach to producing isomaltomegalosaccharide using dextran dextrinase from Gluconobacter oxydans ATCC 11894.

Dextran dextrinase (DDase) catalyzes formation of the polysaccharide dextran from maltodextrin. During the synthesis of dextran, DDase also generates the beneficial material isomaltomegalosaccharide (IMS). The term megalosaccharide is used for a saccharide having DP = 10-100 or 10-200 (DP, degree of polymerization). IMS is a chimeric glucosaccharide comprising α-(1 → 6)- and α-(1 → 4)-linked portions at the nonreducing and reducing ends, respectively, in which the α-(1 → 4)-glucosyl portion originates from maltodextrin of the substrate. In this study, IMS was produced by a practical approach using extracellular DDase (DDext) or cell surface DDase (DDsur) of Gluconobacter oxydans ATCC 11894. DDsur was the original form, so we prepared DDext via secretion from intact cells by incubating with 0.5% G6/G7 (maltohexaose/maltoheptaose); this was followed by generation of IMS from various concentrations of G6/G7 substrate at different temperatures for 96h. However, IMS synthesis by DDext was limited by insufficient formation of α-(1 → 6)-glucosidic linkages, suggesting that DDase also catalyzes elongation of α-(1 → 4)-glucosyl chain. For production of IMS using DDsur, intact cells bearing DDsur were directly incubated with 20% G6/G7 at 45°C by optimizing conditions such as cell concentration and agitation efficiency, which resulted in generation of IMS (average DP = 14.7) with 61% α-(1 → 6)-glucosyl content in 51% yield. Increases in substrate concentration and agitation efficiency were found to decrease dextran formation and increase IMS production, which improved the reaction conditions for DDext. Under modified conditions (20% G6/G7, agitation speed of 100rpm at 45°C), DDext produced IMS (average DP = 14.5) with 65% α-(1 → 6)-glucosyl content in a good yield of 87%. KEY POINTS: • Beneficial IMS was produced using thermostabilized DDase. • Optimum conditions for reduced dextran formation were successfully determined. • A practical approach was established to provide IMS with a great yield of 87%.

Comparative performance of high-rate anaerobic reactors for biodegradation of soybean molasses

Soybean molasses is a residue obtained at the industrial processing of soybean to produce the soy protein concentrate. This residue is the result of the extraction of sugars from de-oiled soybean meal, which viable alternative to its disposal might be the degradation by anaerobic digestion (AD). Thus, a comparative study was performed in two high-rate technologies (UASB and ABR reactors) to evaluate the start-up time and optimum HRT required for the AD of soybean molasses under similar loading operations. The lab-scale reactors were fed with soybean molasses for 134 and 103 days under ambient conditions, respectively. Initially, the UASB reactor was operated at an organic loading rate (OLR) of 0.28 kgCOD m−3 d−1 (HRT = 48 h), while ABR reactor of 0.43 kgCOD m−3 d−1 (HRT = 36 h). OLRs were improved by reducing HRT 48 to 12 h and increasing substrate concentration (COD of 500 to 6000 mg L−1) for six different OLR in both reactors. After the fast start-up period, the UASB reactor attained values of CODTotal and CODFiltered removal efficiencies of 80% and 88%, respectively at an HRT of 24 h (3.91 kgCOD m−3 d−1) whereas ABR reactor presented 95% (CODTotal) and 97% (CODFiltered) at HRT of 36 h (3.86 kgCOD m−3 d−1) for similar OLR. By reducing HRT, both reactors have shown granular sludge flotation (sludge washout) and lower COD removal efficiencies (<80%). The experimental results revealed that high-rate anaerobic configurations (UASB and ABR reactors) can be used for the anaerobic biodegradation of soybean molasses with removal efficiency and a short start-up period.