Slow Hydrolysis Rate Research Articles

Systematic optimization of cellulases production and extraction by Sporotrichum thermophile mutant strain TH3-9 and application in enzymatic hydrolysis of corn stover

Enzymatic hydrolysis of cellulose generally occurs at 40-50 ℃, which can result in slow hydrolysis rate, low sugar yield with incomplete hydrolysis and easy microbial contamination. The limitations can be resolved by adopting cellulases from thermophilic microorganisms. In this work, response surface methodology was used for optimizing cellulases production and extraction by Sporotrichum thermophile mutant strain TH3-9 in solid state fermentation. Results indicated that ammonium sulphate, initial pH and inoculum amount had significant effects on cellulases production. Maximal cellulases activities (endoglucanase activity, 718.03 U/g; filter paper activity, 126.51 U/g; β-glucosidase activity, 444.33 U/g) could be obtained by adopting corn stover (10.0g) and wheat bran (5.0g) as substrates, ammonium sulphate 1.63g, initial pH 5.1 and inoculum amount 13.8% (v/w). Compared with cellulases activities in the initial conditions, optimization resulted in 5.86-fold, 4.29-fold and 6.06-fold increase for endoglucanase activity, filter paper activity and β-glucosidase activity, respectively. During optimization of cellulases extraction, filter paper activity (FPA) was used as the response and results indicated that variables including volume of solvent, rotation speed and time had significant effects on FPA of the crude cellulases solution. Maximal FPA (24.53U/mL) could be obtained while extraction was performed at 30 ℃, 135rpm for 38min with the use of distill water as solvent. In addition, the crude cellulases solution was primarily applied in hydrolysis of alkaline hydrogen peroxide pretreated corn stover and the reducing sugar yield (418.47mg/g) could be obtained during hydrolysis at 50 ℃ for 48h.

Mechanistic insight into the competition between interfacial and bulk reactions in microdroplets through N2O5 ammonolysis and hydrolysis

Reactive uptake of dinitrogen pentaoxide (N2O5) into aqueous aerosols is a major loss channel for NOx in the troposphere; however, a quantitative understanding of the uptake mechanism is lacking. Herein, a computational chemistry strategy is developed employing high-level quantum chemical methods; the method offers detailed molecular insight into the hydrolysis and ammonolysis mechanisms of N2O5 in microdroplets. Specifically, our calculations estimate the bulk and interfacial hydrolysis rates to be (2.3 ± 1.6) × 10−3 and (6.3 ± 4.2) × 10−7 ns−1, respectively, and ammonolysis competes with hydrolysis at NH3 concentrations above 1.9 × 10−4 mol L−1. The slow interfacial hydrolysis rate suggests that interfacial processes have negligible effect on the hydrolysis of N2O5 in liquid water. In contrast, N2O5 ammonolysis in liquid water is dominated by interfacial processes due to the high interfacial ammonolysis rate. Our findings and strategy are applicable to high-chemical complexity microdroplets.

Ascorbic acid reduction pretreatment enhancing metal regulation to improve methane production from anaerobic digestion of waste activated sludge

Conversion of waste activated sludge (WAS) to methane by anaerobic digestion (AD) is often limited by the slow rate of hydrolysis, and the presence of metal ions in sludge is regarded as a critical factor hindering sludge hydrolysis. This study developed a novel strategy to remove Fe from WAS by using ascorbic acid (VC) as a reducing agent under acidic conditions. The feasibility of reduction pretreatment in improving methane production of AD and its intrinsic mechanism were investigated. Results indicate that, under VC doses of 100 mmol/L and pH of 3.50, pretreatment removed 47.60 % of Fe, 59.88 % of Ca, and 51.86 % of Mg contained in the sludge. The removal of metal ions facilitated the disruption of sludge flocculation structure and extracellular polymeric substance (EPS) layers, leading to a 14.78 % increase in cell lysis and a decrease in fractal dimension values to 2.08. Batch AD experiments showed that VC pretreatment improved methane production, with an optimized net methane yield of 190.22 mL/g·VS, an increase of 134.75 % compared to raw WAS. The pretreatment affected the interfacial interaction energy of the sludge, leading to a transformation in the sludge surfaces from hydrophilic to hydrophobic, reducing the interaction between sludge molecules and increasing the number of binding sites available for enzymatic reactions. According to a study of microbial communities, it was found that VC pretreatment caused an increase in the presence of essential functional microbes responsible for hydrolysis, acidification, and methanation. This increase in acetoclastic and hydrogenotrophic methanogens resulted in a substantial enhancement in methane production. These results can be used to develop better pretreatment methods to enhance AD performance.

Local high concentration alkali field accelerated silicon hydrolysis for hydrogen production

Silicon is considered as a promising portable hydrolytic hydrogen production material due to its abundant raw materials, high theoretical capacity (1600 mL/g) and facile storage. However, the low hydrolysis activity and slow hydrolysis rate of silicon hinder its practical application. To address this issue, we introduced CaH2 to form Si-based composites via a flexible ball milling method. The above composites showed ideal yield (1216 mL/g in 10 min), reaction conversion (95.5%) and fast hydrolysis kinetics (225 mL/(s·g)) in deionized water, which is one of the best hydrolysis performances of Si-based composites so far. The hydrolytic process can be divided into two steps: the splitting hydrolysis of CaH2 and the continuous hydrolysis of Si. Combining the dynamic in-situ structural analysis with the analogue simulation, it can be confirmed that the formation of local high concentration alkali field is the key to promote the complete hydrolysis and achieve high yield and fast kinetics of the composites. The existence of lattice defects was proved and the adsorption of Si to hydroxide ion was proved by theoretical calculation. Our work provided a novel idea for the theoretical design of hydrolytic hydrogen production compounds and promoted the practical application of low cost and portable hydrolytic hydrogen production materials.

Anaerobic mono and co-digestion of agro-industrial waste and municipal sewage sludge: Biogas production potential, kinetic modelling, and digestate characteristics

Lignocellulosic substrates, including wastes generated from agro-industrial activities, are bio-residues evaluated to put an end to the energy needs and sustainable waste disposal problems that the world is trying to deal with today. However, the slow rate of hydrolysis of such lignocellulosic wastes limits the biogas production efficiency with the mono-anaerobic digestion (AD) process. Therefore, anaerobic co-digestion (AnCoD) of such wastes with secondary raw materials rich in microorganisms and various nutrients, such as sewage sludge, is an economical and useful method for both enhancing biogas production and disposal of more waste. In this study, AD and AnCoD processes for biogas and methane production were investigated on okra waste (OW) and municipal sewage sludge (MSS) by using cattle manure as inoculation, which constitutes only 8% of the total working volume. Five different substrate mixing ratios (100%OW, 75%OW-25%MSS, 50%OW-50%MSS, 25%OW-75%MSS, and 100MSS%) were prepared to observe the effect of C/N ratio, maximum biogas production potential, methane content, and organic matter removal efficiency on AD and AnCoD processes. All bioreactors were operated in a batch process with 8% TS in mesophilic temperature for 25 days. The amount of biogas produced in the 100%OW experimental set for the AD process was 44% more than 100%MSS. However, the AnCoD process was more efficient than AD, and biogas and methane production increased by up to 66% and 112%, respectively, compared to the mono-substrate experimental set. The highest biogas production value was 1060 mL (375 mL/gVSremoved) with 60% methane content in the bioreactor consisting of 25%OW-75%MSS. As a result of AD and AnCoD process, soluble carbohydrate, protein and COD removal for all bioreactors were between 62% and 79%, 28%-68% and 54%-70%, respectively. The FT-IR peaks seen in digestates were consistent with the presence of functional groups formed as a result of AnCoD. Monod, Cone, Transference function, and Logistic function kinetic models were used for the estimation of cumulative biogas production efficiency. In addition, the energy potential of the methane produced as a result of the AD and AnCoD processes and the financial gain that can be obtained from its direct sale were evaluated.

Abstract EC.03: Distinct Molecular Signatures Of HCM- And DCM-causing Mutations At The Same Residue In The Transducer Of Cardiac Myosin

Mutations in cardiac myosin cause both hypertrophic (HCM) and dilated cardiomyopathy (DCM), and the clinical phenotype ultimately depends on the precise effect of the mutation on the function of myosin. Using recombinant human cardiac myosin, we studied the molecular effects of a novel Q222H mutation in MYH7 found in a patient with severe, early-onset DCM, and compared the effects to the Q222K mutation that was previously reported to cause HCM. The Q222 residue is located within the transducer of the head domain near the ATP binding pocket. The Q222H (DCM) mutant myosin exhibited significantly reduced steady-state actin-activated ATPase activity and a decreased rate of ATP binding in transient kinetic studies, while Q222K (HCM) mutant protein showed an increase in ATPase activity and ATP binding kinetics. In vitro motility assays showed 43% reduction and 32% increase in actin gliding velocity in Q222H and Q222K mutations, respectively. Finally, single nucleotide turnover assays revealed that the fraction of myosin with a slow ATP hydrolysis rate (super relaxed state, or SRX) thought to represent myosin in an autoinhibited state was significantly reduced in Q222K and increased in Q222H. Taken together, the Q222H/K mutations exerted contrasting effects on the motor properties of the cardiac myosin catalytic domain. Cellular characterization and molecular dynamic simulations of cardiac myosin to assess the structural impact of mutant residues are underway. These results highlight how subtle structural differences between cardiomyopathy-causing mutations can lead to significant differences in cardiac myosin motor activity and thus phenotypic expression of cardiomyopathy.

Insights into the fate and properties of organic halamines during ultraviolet irradiation: Implications for drinking water safety

Organic halamines compounds present a significant threat to the safety of drinking water due to their potential toxicity and stability. While Ultraviolet (UV) disinfection is commonly used for water treatment, its specific effects on organic halamines and the underlying mechanisms remain poorly understood. In this study, we investigated eight amino acid-derived organic chlor- and bromamines as representative compounds. Our findings revealed that organic halamines have a slow hydrolysis rate (<10−3 M−1 s−1) and can persist in water for extended periods (30–2000 min). However, their disinfection efficacy against Staphylococcus aureus and their ability to degrade micropollutants like carbamazepine were found to be limited. Interestingly, under UV irradiation, the N-X bonds in organic halamines were observed to break, leading to accelerated decomposition and the generation of abundant free radicals. These free radicals synergistically facilitated the removal of micropollutants and the inactivation of pathogenic microorganisms. It is worth noting that this transformation of organic halamines during UV disinfection resulted in a slight increase in the concentrations of nitrogenous disinfection byproducts. These findings shed light on the behavior and characteristics of organic halamines during UV disinfection processes, providing crucial insights for effectively managing drinking water quality impacted by these compounds. By understanding the implications of organic halamines, we can refine water treatment strategies and ensure the safety of drinking water supplies.

Brush border enzyme hydrolysis and glycaemic effects of isomaltulose compared to other saccharides in dogs.

Digestible carbohydrates differ in glycaemic response, therewith having the potential to influence metabolic conditions such as insulin resistance and diabetes mellitus. Isomaltulose has been proven to lower the glycaemic response in humans, which to date has not been studied in dogs. Therefore, the aim of the present study was to characterise the digestibility, as well as the physiological effects of isomaltulose in dogs, in comparison to other saccharides. To this end, three studies were performed. Study 1 was an in vitro study, evaluating the small intestinal hydrolysis of isomaltulose compared to other relevant carbohydrate sources. Three of these saccharides, having close and low-moderate degrees of hydrolysis by brush border enzymes, were also evaluated in vivo for their glycaemic effects by measuring plasma levels of glucose, insulin and glucagon-like peptide 1 (GLP-1) 0-180 min after administration of a single dosage after an overnight fast (i.e., isomaltulose, sucrose and maltodextrin in a 3 × 3 Latin-square design, in 9 dogs, Study 2). To understand if digestive enzymes, underlying glycaemic responses for isomaltulose and sucrose can be upregulated, we exposed dogs to these saccharides for 2 weeks and repeated the measurements after an overnight fast in 18 dogs (Study 3). Isomaltulose was hydrolysed by intestinal enzyme preparation from all three dogs, but the degrading activity was low (e.g., 3.95 ± 1.03 times lower vs. sucrose), indicating a slower rate of hydrolysis. Isomaltulose had a low glycaemic response, in line with in vitro data. In vitro hydrolysis of sucrose was comparable or even higher than maltodextrin in contrast to the more pronounced glycaemic response to maltodextrin observed in vivo. The numerically higher blood glucose response to sucrose after continuous consumption, might indicate an adaptive response. In conclusion, the current work provides valuable insights into the digestion physiology of various saccharides in dogs. Further investigations on related benefits are thus warranted.

Effect and mechanism analysis of dual enzyme pretreatment on enhancement of volatile fatty acids production in kitchen waste anaerobic acidification

The high content of cellulose and hemicellulose in kitchen waste (KW) results in a slow hydrolysis rate and subsequently hinders the yields of volatile fatty acids (VFAs) in anaerobic acidification. In this study, cellulase and hemicellulase were used for the pretreatments of KW to tackle the problem of difficult hydrolysis in anaerobic acidification. The results indicated that the joint use of double enzymes significantly improved the hydrolysis of cellulose and hemicellulose during pretreatment, thus enhancing the VFAs production in anaerobic acidification. The highest production of TVFAs was 50.73 ± 2.10 g COD/L and it was improved by 62.77% compared with the unpretreated group. In addition, the 16S rRNA amplicon sequencing analysis showed that the relative abundance of Firmicutes related with the VFAs production increased from 17.77% to 50.55%. The potential metabolism function analysis (PICRUSt2) indicated that the enrichment of glyceraldehyde-3-phosphate dehydrogenase (EC:1.2.1.9) and acetyl-CoA C-acetyltransferase (EC:2.3.1.9) contributed to the production of VFAs in AD. The results suggested that collaborative enzyme pretreatment could be one of the effective methods for the hydrolysis of cellulose and hemicellulose in KW and provided constructive mechanistic insights into recovering valuable products from KW.

Molecular characterization of a novel MYH7 mutation Q222H in a patient with severe dilated cardiomyopathy.

Mutations in the cardiac myosin heavy chain gene (MYH7) are known to cause both hypertrophic and dilated cardiomyopathy (HCM and DCM), and the use of recombinant human cardiac myosin has enabled the study of the precise biochemical and biophysical effects of these mutations on myosin function at the molecular level. Here we report a case of a patient with severe, early-onset DCM who carries a previously unreported Q222H mutation in MYH7. This mutation affects a residue at the distal end of the ATP binding pocket of cardiac myosin. The Q222H mutant myosin exhibited significantly reduced steady-state actin-activated ATPase activity and a decreased rate of ATP binding in transient kinetic studies. It also showed a ∼50% reduction in actin gliding velocity in an in vitro motility assay. Single-nucleotide turnover assays were performed comparing Q222H mutant myosin 2-headed constructs with either a long S2 tail capable of forming a folded-back auto-inhibited state or a short S2 tail that cannot stably fold back and sequester heads. These studies revealed that the fraction of myosin with a slow ATP hydrolysis rate (super relaxed state, or SRX) thought to represent myosin in an autoinhibited state did not differ significantly between WT and the Q222H mutant myosins. Taken together, the Q222H mutation exerts substantial effects on the motor properties of the cardiac myosin catalytic domain, rather than affecting the availability of myosin heads in the sarcomere. Further transient kinetic assays are underway to delineate precise steps in the actin-myosin ATPase cycle affected by the mutation. Future work also includes comparing these effects with the Q222K mutation that has been reported to cause HCM, to gain further insights into the molecular basis for phenotypic determination of cardiomyopathy.

Potential of anaerobic co-digestion of acidic fruit processing waste and waste-activated sludge for biogas production

Abstract The potential of acidic fruit processing waste (FPW) and waste-activated sludge (WAS) co-digestion for methane production was investigated using batch and continuous experiments. First, batch experiments showed the co-digestion of FPW and WAS solved acid accumulation and increased cumulative biogas production. When the volatile solid (VS) ratio of FPW to WAS was 2:1, the cumulative biogas production was the highest (4,695.47 mL), which increased by 5.2% and 10.5% compared with the VS ratios of 3:1 and 1:1, respectively. Methane production was the rate-limiting step when the FPW percentage was high, and hydrolysis was the rate-limiting step when the WAS percentage was high. Second, the continuous experiments showed fermentation was the most stable when the VS ratio was 2:1, without acid accumulation or excessive total alkalinity consumption. Additionally, the ammonia nitrogen content in the system was above 138.34 mg·L−1, which solved the slow WAS hydrolysis rate and low nitrogen supply efficiency. Microbial community structure of the reactor was correlated with substrate composition greatly. On the 180th day, the relative abundance levels of Bacteroides, Paludibacter, Litorilinea, Levilinea, and Smithella were higher than those on the 120th day and the 240th day. The enrichment of those bacterial groups was beneficial to improve the substrate hydrolysis rate and reduce the influence of organic acids on the anaerobic system.

Enhanced two-phase anaerobic digestion of waste activated sludge by combined free nitrous acid and manganese dioxide

Anaerobic digestion (AD) is a mature and reliable technology for sludge treatment, but it still faces many technical problems including slow hydrolysis rate and low methane yield. Free nitrous acid (FNA) pretreatment was previously confirmed in enhancing sludge hydrolysis and extracellular polymeric substances (EPS) disruption in anaerobic digestion. It was hypothesized that, due to the manganese dioxide (MnO2) addition during methanogenic stage, direct interspecies electron transfer (DIET) facilitated the biological activities, thus boosting the methane production in FNA-pretreated two-phase AD model. The results showed that the methane production of the combination of FNA pretreatment and MnO2 addition was improved by 18.64% and 22.23%, compared with reactors that solely treated by either MnO2 addition or FNA pretreatment. The changes of microbial metabolism activity were evaluated by measuring the coenzyme F420 and electron transfer system (ETS). The activities of coenzyme F420 and ETS were increased to 156.26% and 134.71% in two-phase AD model, respectively. Meanwhile, the obvious microbial community succession was found with the enrichment of methanogens such as Methanosarcina and Methanobacterium. Overall, the combination of FNA pretreatment and MnO2 addition avoided the inhibition of FNA pretreatment on methanogenesis in the early stage, and showed positively synergistic effect on methane production. The enhancement of microbial metabolism was responsible for, promoting methane production in the two-stage AD model. This research provides an alternative strategy for efficiency improvement of anaerobic sludge digestion as well as methane production.

Structures and mechanisms of actin ATP hydrolysis

The major cytoskeleton protein actin undergoes cyclic transitions between the monomeric G-form and the filamentous F-form, which drive organelle transport and cell motility. This mechanical work is driven by the ATPase activity at the catalytic site in the F-form. For deeper understanding of the actin cellular functions, the reaction mechanism must be elucidated. Here, we show that a single actin molecule is trapped in the F-form by fragmin domain-1 binding and present their crystal structures in the ATP analog-, ADP-Pi-, and ADP-bound forms, at 1.15-Å resolutions. The G-to-F conformational transition shifts the side chains of Gln137 and His161, which relocate four water molecules including W1 (attacking water) and W2 (helping water) to facilitate the hydrolysis. By applying quantum mechanics/molecular mechanics calculations to the structures, we have revealed a consistent and comprehensive reaction path of ATP hydrolysis by the F-form actin. The reaction path consists of four steps: 1) W1 and W2 rotations; 2) PG-O3B bond cleavage; 3) four concomitant events: W1-PO3- formation, OH- and proton cleavage, nucleophilic attack by the OH- against PG, and the abstracted proton transfer; and 4) proton relocation that stabilizes the ADP-Pi-bound F-form actin. The mechanism explains the slow rate of ATP hydrolysis by actin and the irreversibility of the hydrolysis reaction. While the catalytic strategy of actin ATP hydrolysis is essentially the same as those of motor proteins like myosin, the process after the hydrolysis is distinct and discussed in terms of Pi release, F-form destabilization, and global conformational changes.

What Is the Right Rate? Determining Digestibility Kinetics of Pretreated Waste Activated Sludge During Anaerobic Digestion

Pretreatment of municipal sludge has the potential to improve anaerobic digestion by increasing the ultimate biodegradability of particulate chemical oxygen demand (PCOD) to methane (CH4) gas. However, the effect of pretreatment, that is, increased soluble COD from solubilization of PCOD, makes it difficult to correctly estimate hydrolysis kinetics in biochemical methane potential tests. In this study, batch digestion tests were used to evaluate the general impact of alkaline and thermal pretreatment of waste activated sludge (WAS) and to determine a different method to estimate hydrolysis rates from pretreated streams. Throughout the 60-day batch experiments, liquid and gas samples were collected and analyzed for total (TCOD) and semi-soluble (SSCOD) COD, total (TSS) and volatile (VSS) suspended solids, pH, volatile fatty acids (VFAs), and ammonium concentration. Pretreatments significantly decreased VSS concentrations at the start of digestion, but all samples showed similar VSS destruction to the control at the end of the experiments. Ultimate CH4 production was significantly increased by alkaline and thermal pretreatments, which produced 203 and 219 mL CH4 gVSS−1, respectively, versus 175 mL CH4 gVSS−1 in the control. Although CH4 generation was a good proxy for solids hydrolysis of untreated WAS, CH4 generation significantly overestimated the rate of hydrolysis of pretreated solids. The overestimation was due to the abundance of SSCOD available at the start of the experiment being converting to CH4. Incorporating SSCOD or VFAs into the hydrolysis kinetics resulted in much slower hydrolysis rates that were similar to those measured by VSS destruction directly (e.g., 0.08 d−1 with SSCOD + CH4 vs. 0.19 d−1 from CH4 alone for thermal pretreatment vs. 0.05 d−1 with VSS alone). The slower kinetics indicate that the necessity to understand the trade-offs between the performance advantages of pretreatment (i.e., solids destruction and energy conversion) versus the operational and financial impacts on large-scale digestion.

Impacts of Anaerobic Co-Digestion on Different Influencing Parameters: A Critical Review

Lignocellulosic feedstocks are year-round, available bio-residues that are the right candidates for counteracting the energy crises and global warming facing the world today. However, lignin leads to a slow hydrolysis rate and is a major bottleneck for biogas production via anaerobic digestion. Anaerobic co-digestion (AcoD) is an economical method available, which overcomes the limitation of a single feedstock’s properties in an anaerobic digestion process. This paper critically reviews the impacts of co-digestion on lignocellulosic biomass degradation, process stability, various working parameters, and microbial activities that improve methane yields. A combination of compatible substrates is chosen to improve the biomethane yield and conversion rate of organic matter. AcoD is a promising method in the delignification of lignocellulosic biomass as an acid pretreatment. Ultimate practices to control the impact of co-digestion on system performances include co-feed selection, in terms of both carbon-to-nitrogen (C/N) and mixing ratios, and other operating conditions. A detailed analysis is performed using data reported in the recent past to assess the sensitivity of influencing parameters on the resultant biogas yield. For the investigators motivated by the basic principles of AcoD technology, this review paper generates baseline data for further research work around co-digestion.

Principles and potential of thermal hydrolysis of sewage sludge to enhance anaerobic digestion

Sewage sludge is rich source of carbon, nutrients, and trace elements and can be subjected to proper treatment before disposal to fulfill government legislation and protect receiving environments. Anaerobic digestion (AD) is a well-adopted technology for stabilizing sewage sludge and recovering energy-rich biogas and nutrient-rich digestate. However, a slow hydrolysis rate limits the biodegradability of sludge. In the present study we have attempted to explain the potential of thermal hydrolysis to enhance anaerobic digestion of sewage sludge. Thermal pretreatment improves biodegradability and recycling of the sludge as an excellent energy and nutrients recovery source at reasonable capital (CAPEX) and operational (OPEX) costs. Other pretreatments like conventional (below/above 100 °C), temperature-phased anaerobic digestion (TPAD), microwave and chemically mediated thermal pretreatment have also been accounted. This review provides a holistic overview of sludge's characterization and value-added properties, various techniques used for sludge pretreatment for resource recovery, emphasizing conventional and advanced thermal pretreatment, challenges in scale-up of these technologies, and successful commercialization of thermal pretreatment techniques.

Impact of fat replacement by a hybrid gel (canola oil/candelilla wax oleogel and gelatinized corn starch hydrogel) on dough viscoelasticity, color, texture, structure, and starch digestibility of sugar-snap cookies

The reduction of fat content is an important issue in the formulation of bakery products. In this work, the fat content in sugar-snap cookies was reduced by replacing a commercial shortening with a hybrid gel (HG). The HG was formed by mixing a canola oil/candelilla oleogel (5 g/100 g) and a gelatinized corn starch hydrogel (5 g/100 g) in a 1:1 ratio. The commercial shortening was replaced (0, 25, 50, 75, and 100%) by HG, achieving a fat content reduction of up to 50%. HG cookies exhibited reduced hardness and improved firmness, their surface color pattern exhibited an increased yellowness and lightness as HG increased. FTIR analysis revealed that the starch contained in the HG cookies was more hydrated and unordered, and the secondary structure of the proteins was less ordered. The starch hydrolysis pattern exhibited two phases that reflected structures with fast and slow hydrolysis rates. As a consequence of the gelatinized starch fraction, the HG cookies showed increased starch digestibility, with the rapidly digestible starch varying from 19.0% for the control cookie to about 43% for the HG cookie with complete shortening replacement. Overall, hybrid gels offer an alternative to commercial shortening use for formulating sugar-snap cookies with lower saturated fat content.

Effects of pH Adjustment on the Release of Carbon Source of Particulate Organic Matter (POM) in Domestic Sewage

The use of anaerobic hydrolytic fermentation to develop more available carbon sources from domestic sewage influent particulate organic matter (POM) has received increasing attention. However, the slow hydrolysis rate of POM limits the application of this technology. This study aimed to improve the carbon source release efficiency of POM by pH adjustment and to reveal the hydrolysis mechanism. Results showed that adjusting the initial pH of POM to 3, 9, and 11 enhanced carbon source release in the anaerobic hydrolysis fermentation process of POM. The pretreatment under pH value of 11 contributed to the highest yield and productivity of carbon source, reaching the soluble chemical oxygen demand (SCOD) of 2782 mg/L at the 4th day. The pH 3 pretreatment was more beneficial for phosphorus resource recovery, which contributed to the highest release concentration of PO43−-P, reaching 48.2 mg/L at the 3rd day, accounting for 90% of TP. Microbial community structure analysis indicated that pH 11 preconditioning promoted the enrichment of proteolytic bacteria (Proteocatella and Proteiniclasticum) and polysaccharide hydrolytic bacteria (Trichococcus and Acinetobacter) and inhibited the growth of acetate-consuming methanogenic archaea, which contributed to the highest carbon release of POM in domestic sewage.

Rational Design, Synthesis, and Mechanism of (3S,4R)-3-Amino-4-(difluoromethyl)cyclopent-1-ene-1-carboxylic Acid: Employing a Second-Deprotonation Strategy for Selectivity of Human Ornithine Aminotransferase over GABA Aminotransferase.

Human ornithine aminotransferase (hOAT) is a pyridoxal 5'-phosphate (PLP)-dependent enzyme that contains a similar active site to that of γ-aminobutyric acid aminotransferase (GABA-AT). Recently, pharmacological inhibition of hOAT was recognized as a potential therapeutic approach for hepatocellular carcinoma. In this work, we first studied the inactivation mechanisms of hOAT by two well-known GABA-AT inactivators (CPP-115 and OV329). Inspired by the inactivation mechanistic difference between these two aminotransferases, a series of analogues were designed and synthesized, leading to the discovery of analogue 10b as a highly selective and potent hOAT inhibitor. Intact protein mass spectrometry, protein crystallography, and dialysis experiments indicated that 10b was converted to an irreversible tight-binding adduct (34) in the active site of hOAT, as was the unsaturated analogue (11). The comparison of kinetic studies between 10b and 11 suggested that the active intermediate (17b) was only generated in hOAT and not in GABA-AT. Molecular docking studies and pKa computational calculations highlighted the importance of chirality and the endocyclic double bond for inhibitory activity. The turnover mechanism of 10b was supported by mass spectrometric analysis of dissociable products and fluoride ion release experiments. Notably, the stopped-flow experiments were highly consistent with the proposed mechanism, suggesting a relatively slow hydrolysis rate for hOAT. The novel second-deprotonation mechanism of 10b contributes to its high potency and significantly enhanced selectivity for hOAT inhibition.

Effect of the addition of different sodium alginates on viscoelastic, structural features and hydrolysis kinetics of corn starch gels

Corn starch gels (1:4 w:w) were made with the addition (up to 2% w/w starch basis) of sodium alginates of different average molecular sizes and nature. Rheology of the starchy gels were characterized to evaluate their viscoelasticity. Gels were subjected to in vitro digestion to estimate the effect of alginates addition on glycemic response. Tested alginates significantly decreased the value of the elastic modulus of gels. The coordination number obtained from complex modulus indicated that in general, less compact structures were obtained in the presence of alginates. These results were confirmed by scanning electron micrographs, where significant differences in the structure of alginate-corn starch gels were observed, but molecular size of alginates was not critical. Hydrolysis rate decreased with the presence of alginates in corn starch gels by interactions of alginates with leached amylose. At high alginate content, slower hydrolysis rates were determined when the most hygroscopic alginates were added.