Recent advances in biosynthesis mechanisms and yield enhancement strategies of erythritol.

Erythritol is a four-carbon sugar alcohol naturally produced by microorganisms as an osmoprotectant. As a new sugar substitute, erythritol has recently been popular on the ingredient market because of its unique nutritional characteristics. Even though the history of erythritol biosynthesis dates from the turn of the twentieth century, scientific advancement has lagged behind other polyols due to the relative complexity of making it. In recent years, biosynthetic methods for erythritol have been rapidly developed due to an increase in market demand, a better understanding of metabolic pathways, and the rapid development of genetic engineering tools. This paper reviews the history of industrial strain development and focuses on the underlying mechanism of high erythritol production by strains gained through screening or mutagenesis. Meanwhile, we highlight the metabolic pathway knowledge of erythritol biosynthesis in microorganisms and summarize the metabolic engineering and research progress on critical genes involved in different stages of the synthetic pathway. Lastly, we talk about the still-contentious issues and promising future research directions that will help break the erythritol production bottleneck and make erythritol production greener and more sustainable.

Role of STAT3 and NRF2 in Tumors: Potential Targets for Antitumor Therapy.

Signal transducer and activator of transcription 3 (STAT3) and nuclear factor erythroid-derived 2-like 2 (NRF2, also known as NFE2L2), are two of the most complicated transcription regulators, which participate in a variety of physiological processes. Numerous studies have shown that they are overactivated in multiple types of tumors. Interestingly, STAT3 and NRF2 can also interact with each other to regulate tumor progression. Hence, these two important transcription factors are considered key targets for developing a new class of antitumor drugs. This review summarizes the pivotal roles of the two transcription regulators and their interactions in the tumor microenvironment to identify potential antitumor drug targets and, ultimately, improve patients' health and survival.

Batch fermentation kinetics of acetoin produced by Bacillus subtilis HB-32.

OBJECTIVES: The aim of this work was to study the changes of bacterial cell growth, acetion formation and glucose consumption with fermentation time during batch cultivation. RESULTS: A mathematical model of cell growth, product synthesis, and substrate consumption changes with time during the batch cultivation of acetion was established. By analyzing the fitting curve of the kinetic model, it is found that the calculated value of the model fits well with the experimental value, and the fitting model R2 is greater than 0.98. CONCLUSIONS: The kinetic model established in this experiment can better reflect the batch cultivation process of acetion.

2-Keto-D-gluconic acid and prodigiosin producing by a Serratia marcescens.

Microbial fermentation has become the main method to produce target compound. In this study, a 2-Keto-D-gluconic acid (2-KGA) producing mutant strain was obtained by mutation with rational screening methods. Meanwhile, prodigiosin was produced when the nitrogen source in the medium was changed to peptone and its fermentation conditions were evaluated to achieve high-efficient accumulation. The mutant strain SDSPY-136 was firstly identified as Serratia marcescens by morphological observation and 16S rDNA sequencing. The 2-KGA synthetic capacity of S. marcescens SDSPY-136 was evaluated by shake fermentation with 110 g/L glucose as substrates. For fermentation, 2-KGA yield, conversation rate and purity of SDSPY-136 reached 104.60 g/L, 95.10%, 99.11% in 72 h. The red pigment was extracted from the fermentation broth using acidic methanol and identified as prodigiosin by FT-IR. The optimal conditions were as follows: glycerol 20 g/L, peptone 20 g/L, MgSO415 g/L, pH 6.0, a 2% (v/v) inoculum, 30 °C and 200 rpm of shaking culture. Eventually, prodigiosin reached a yield of 9.89 g/Lafter shake culturing for 50 h under this condition. The mutant S. marcescens SDSPY-136 was shown to be promising for 2-KGA and prodigiosin production and a suitable object for prodigiosin metabolism research of S. marcescens.

Epidemic features of seasonal influenza transmission among eight different climate zones in Gansu, China.

BACKGROUNDS: Seasonal influenza remains epidemic globally with a substantial health burden. Understanding the transmission patterns and epidemic features of influenza may facilitate the improvement of preventive and control measures. This study aims to assess the epidemic features of influenza among different climate zones and identify high-risk zones across Gansu province, China. METHODS: We collected weekly influenza cases at county-level between 1st January 2012 and 31st December 2016, as well as climate zones classification shapefile data from Köppen-Geiger climate map. We compared the epidemic features (Frequency index (α), Duration index (ß) and Intensity index (γ)) of influenza among different climate zones. Spatial cluster analysis was used to examine the high-risk areas of transmission of influenza. RESULTS: The distribution of cases existed significant differences among eight climate zones (F-test: 267.02, p < 0.05). The highest mean weekly incidence rate (per 100,000 population) was 0.59 in snow climate with dry winter and warm summer (Dwb). The primary (relative risk (RR): 3.61, p < 0.001) and secondary (RR: 2.45, p < 0.001) clusters were located in Dwb. The highest values of α, ß and γ were 1.00, 261 and 154.38 in Dwb. The hot spots (high-high clusters) of the epidemic indices were detected in Dwb. CONCLUSIONS: This study found the variability of epidemic features of influenza among eight climate zones. We highlight that Dwb was the high-risk zone where influenza clustered with the highest incidence rate and epidemic temporal indices. This provide further insight into potential improvement of preventive measures by climate zones to minimize the impact of epidemics.

Enhanced itaconic acid production in Yarrowia lipolytica via heterologous expression of a mitochondrial transporter MTT.

Itaconic acid, a promising platform chemical, has been applied in many fields of industrial production. As a potential candidate for itaconic acid production, Yarrowia lipolytica possesses several innate abilities such as the tolerance of low-pH and high-shear stress, fast growth rate, cultivation flexibility, and easy for genetic manipulation. Here, Y. lipolytica Po1f which was tested to show high tolerance to itaconic acid could accumulate itaconic acid (0.363 g/L) by expressing the Aspergillus terreus cis-aconitic acid decarboxylase (CAD). Then, we tried to improve the supply and transport of the immediate precursor cis-aconitic acid by overexpressing a series of genes; these results indicate that overexpression of mitochondrial cis-aconitate transporter MTT is beneficial to the itaconic acid biosynthesis in Y. lipolytica. Further culture optimization enabled 22.03 g/L of itaconic acid to be produced in bioreactors, about 60-fold improvement over the initial titer, which is the highest itaconic acid production achieved at low pH by yeast reported worldwide, to data. This study demonstrates the great potential of Y. lipolytica as an industrial platform for itaconic acid production.

Composition and Rheological Properties of Polysaccharide Extracted from Tamarind (Tamarindus indica L.) Seed.

A polysaccharide was extracted in high yield from tamarind (Tamarindus indica L.) seed (TSP) by acidic hot water extraction and ethanol precipitation. It was composed of 86.2% neutral polysaccharide, 5.4% uronic acid and 1.3% protein. The molecular weight of TSP was estimated to be about 1735 kDa, with glucose, xylose, and galactose in a molar ratio of 2.9:1.8:1.0 as the major monosaccharides. The steady shear and viscoelastic properties of TSP aqueous solutions were investigated by dynamic rheometry. Results revealed that TSP aqueous solution at a concentration above 0.5% (w/v) exhibited non-Newtonian shear-thinning behavior. Dynamic oscillatory analysis revealed that 10% (w/v) TSP showed as a "weak gel" structure. Apparent viscosities and viscoelastic parameters of TSP solutions decreased drastically in an alkaline solution of pH > 10, but slightly influenced by acidic solution, high temperature and the presence of salt ions and sucrose. These results indicated that TSP possessed excellent pH-resistance and thermo-stability, which might be suitable for applications in acidic beverages and high-temperature processed foodstuffs.

Heterodimerization of the kappa opioid receptor and neurotensin receptor 1 contributes to a novel ß-arrestin-2-biased pathway.

Together with its endogenous ligands (dynorphin), the kappa opioid receptor (KOR) plays an important role in modulating various physiological and pharmacological responses, with a classical G protein-coupled pathway mediating analgesia and non-G protein-dependent pathway, especially the ß-arrestin-dependent pathway, eliciting side effects of dysphoria, aversion, drug-seeking in addicts, or even relapse to addiction. Although mounting evidence has verified a functional overlap between dynorphin/KOR and neurotensin/neurotensin receptor 1 (NTSR1) systems, little is known about direct interaction between the two receptors. Here, we showed that KOR and NTSR1 form a heterodimer that functions as a novel pharmacological entity, and this heterodimer, in turn, brings about a switch in KOR-mediated signal transduction, from G protein-dependent to ß-arrestin-2-dependent. This was simultaneously verified by analyzing a KOR mutant (196th residue) that lost the ability to dimerize with NTSR1. We also found that dual occupancy of the heterodimer forced the ß-arrestin-2-dependent pathway back into Gi protein-dependent signaling, according to KOR activation. These data provide new insights into the interaction between KOR and NTSR1, and the newly discovered role of NTSR1 acting as a switch between G protein- and ß-arrestin-dependent pathways of KOR also suggests a new approach for utilizing pathologically elevated dynorphin/KOR system into full play for its analgesic effect with limited side effects.

Effect of deletion of 2,3-butanediol dehydrogenase gene (bdhA) on acetoin production of Bacillus subtilis.

The present work aims to block 2,3-butanediol synthesis in acetoin fermentation of Bacillus subtilis. First, we constructed a recombinant strain BS168D by deleting the 2,3-butanediol dehydrogenase gene bdhA of the B. subtilis168, and there was almost no 2,3-butanediol production in 20 g/L of glucose media. The acetoin yield of BS168D reached 6.61 g/L, which was about 1.5 times higher than that of the control B. subtilis168 (4.47 g/L). Then, when the glucose concentration was increased to 100 g/L, the acetoin yield reached 24.6 g/L, but 2.4 g/L of 2,3-butanediol was detected at the end of fermentation. The analysis of 2,3-butanediol chiral structure indicated that the main 2,3-butanediol production of BS168D was meso-2,3-butanediol, and the bdhA gene was only responsible for (2R,3R)-2,3-butanediol synthesis. Therefore, we speculated that there may exit another pathway relating to the meso-2,3-butanediol synthesis in the B. subtilis. In addition, the results of low oxygen condition fermentation showed that deletion of bdhA gene successfully blocked the reversible transformation between acetoin and 2,3-butanediol and eliminated the effect of dissolved oxygen on the transformation.

Identification of serine 348 on the apelin receptor as a novel regulatory phosphorylation site in apelin-13-induced G protein-independent biased signaling.

Phosphorylation plays vital roles in the regulation of G protein-coupled receptor (GPCR) functions. The apelin and apelin receptor (APJ) system is involved in the regulation of cardiovascular function and central control of body homeostasis. Here, using tandem mass spectrometry, we first identified phosphorylated serine residues in the C terminus of APJ. To determine the role of phosphorylation sites in APJ-mediated G protein-dependent and -independent signaling and function, we induced a mutation in the C-terminal serine residues and examined their effects on the interaction between APJ with G protein or GRK/ß-arrestin and their downstream signaling. Mutation of serine 348 led to an elimination of both GRK and ß-arrestin recruitment to APJ induced by apelin-13. Moreover, APJ internalization and G protein-independent ERK signaling were also abolished by point mutation at serine 348. In contrast, this mutant at serine residues had no demonstrable impact on apelin-13-induced G protein activation and its intracellular signaling. These findings suggest that mutation of serine 348 resulted in inactive GRK/ß-arrestin. However, there was no change in the active G protein thus, APJ conformation was biased. These results provide important information on the molecular interplay and impact of the APJ function, which may be extrapolated to design novel drugs for cardiac hypertrophy based on this biased signal pathway.

Optimization of fermentation medium for acetoin production by Bacillus subtilis SF4-3 using statistical methods.

To improve the acetoin-producing ability of Bacillus subtilis SF4-3, isolated from "natto," a Japanese traditional food, the fermentation medium was optimized in shake-flask fermentation by statistically designed methods. Based on results of the single-factor experiment, orthogonal experiment, and Plackett-Burman design, yeast extract, corn steep liquor, and urea were identified as showing significant influence on the acetoin production. Subsequently, the optimum combination of the three factors was investigated by the Box-Behnken design (BBD) of response surface methodology (RSM) in order to further enhance the acetoin production. The maximum acetoin yield of 45.4 g/L was predicted when the concentrations of yeast extract, corn steep liquor, and urea were 8.5 g/L, 14.6 g/L, and 3.8 g/L, respectively. The results were further confirmed in triplicate experiments using the optimized medium (glucose 160 g/L, yeast extract 8.5 g/L, corn steep liquor 14.6 g/L, urea 3.8 g/L, manganese sulfate 0.05 g/L, ferrous sulfate 0.05 g/L), and an acetoin yield of 46.2 g/L was obtained in the validation experiment, which was in agreement with the prediction. After the optimization of medium components, an increase of 36.28% in acetoin production was achieved in comparison to that at the initial medium levels.

Isolation of acetoin-producing Bacillus strains from Japanese traditional food-natto.

In this study, Bacillus strains with an ability to produce acetoin were isolated from a Japanese traditional food, natto, on the basis of the Voges-Proskauer (VP) reaction, and strain SF4-3 was shown to be a predominant strain in acetoin production. Based on a variety of morphological, physiological, and biochemical characteristics as well as the nucleotide sequence analysis of 16S rDNA, the strain SF4-3 was identified as Bacillus subtilis. When it was incubated at 37°C with a speed of 180 rpm for 96 hr in the flasks, the maximum acetoin concentration was up to 33.90 g/L. The fermentation broths were determined by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) analyses; the results showed that the major metabolite was acetoin, and the purity could reach more than 95% without butanedione and 2,3-butanediol, which were usually produced together with acetoin in other strains. A novel aqueous two-phase system (ATPS) composed of hydrophilic solvents and inorganic salts was developed for the extraction of acetoin from fermentation broths. The ethanol and dipotassium hydrogen phosphate system could be used to extract acetoin from fermentation broths. The influences of phase composition on partition of acetoin were investigated. The maximum partition coefficient (9.68) and recovery (94.6%) of acetoin were obtained, when 25% (w/w) dipotassium hydrogen phosphate and 24% (w/w) ethanol were used.

Rheological Properties and Kinetics of Gelation of Binary Polymers between Xanthan Gum and Locust Bean Gum.

The synergistic interaction and gelling kinetics between xanthan gum (XG) and locust bean gum (LBG) at different mass ratios (XG/LBG 9:1, 7:3, 5:5, 3:7, 1:9) were investigated using a rheometer. The results showed that the mixtures of XG and LBG induced gel formation, and the strongest gel structure was found for the mixture of XG/LBG 3:7 according to the yield stress, storage modulus (G'), and power law parameters. Temperature ramp studies indicated that heating destroyed the gels at 55~60 °C, while cooling induced the sol-gel transition at around 52 °C for all mixtures. Structure developing rate (SDR) curves showed that XG/LBG 3:7 exhibited the highest SDR during the cooling ramp among all the samples. Non-isothermal kinetic analysis demonstrated that the gelation process of XG/LBG mixtures during cooling included two steps: a high-temperature region (55~39 °C) needing higher activation energy (Ea, 111.97 to 199.20 kJ/mol for different mixtures) and a low-temperature region (39~20 °C) needing lower Ea (74.33 to 85.31 kJ/mol), which indicated higher energy barriers to overcome at the initial stage of gel formation. The lowest Ea of 74.33 kJ/mol was found for XG/LBG 3:7 in the low-temperature region. Scanning electron microscopy (SEM) showed that the gel of XG/LBG 3:7 presented the densest entanglements. These results indicated the strongest synergism interaction occurred in XG/LBG 3:7 to form gel network structures. This study will help promote the application of XG-LBG blends to design novel food structures.

The regulatory effects of the apelin/APJ system on depression: A prospective therapeutic target.

Depression is a debilitating neuropsychological disorder characterized by high incidence, high recurrence, high suicide, and high disability rates, which poses serious threats to human health and imposes heavy psychological and economic burdens on family and society. The pathogenesis of depression is extremely complex, and its etiology is multifactorial. Mounting evidence suggests that apelin and apelin receptor APJ, which compose the apelin/APJ system, are related to the development of depression. However, the specific mechanism is still unclear, and research in this area in human is still insufficient. Acceleration of research into the regulatory effects and underlying mechanisms of the apelin/APJ system in depression may identify attractive therapeutic targets and contribute to the development of novel intervention strategies against this devastating psychological disorder. In this review, we mainly discuss the regulatory effects of apelin/APJ system on depression and its potential therapeutic applications.

Interactions between structure and function of resistant glucans for alleviating type 2 diabetes mellitus (T2DM) and its complications in mice.

Resistant glucan, a functional dietary fiber, has been shown to alleviate type 2 diabetes mellitus (T2DM) and its complications in clinical studies. However, the interactions between the special structure of resistant glucan and the metabolism-related pathways in T2DM have not yet been systematically studied. This study identified the structural differences between resistant glucans prepared by new and old methods. Oral gavage with two resistant glucans in T2DM mice, led to significant improvements in glucose and lipid metabolism as measured by related indicators (including gut microbiota, fecal metabolites, and physiological and biochemical indexes). According to these results, in addition to van der Waals forces, micelle formation, and hydrogen bonding, the branching structures of resistant glucans produced more hydroxyl, carbonyl, and keto groups that linked cholesterols, cholesterol esters, and low-density lipoprotein intermediates. Moreover, after lipid clearing, the metabolic environment was more conducive to the proliferation of specific gut microbiota (including Phascolarctobacterium, Prevotella, Butyricicoccus, Weissella, and Anaerostipes) with decreasing abundance ratios of Firmicutes and Bacteroides. This facilitated the synthesis of high-density lipoprotein, conversion of cholesterol into coprostanol, and production of short-chain fatty acids and bile acids. Our findings provide a foundation for comprehensive investigation of the structure of resistant glucan in the promotion and prevention of T2DM.

Preparation of resistant glucan by high pressure processing (HPP) and enzymatic methods increases indigestibility.

This study describes a new method for producing high-quality resistant glucan by characterizing the structural mechanism of indigestibility. The structures and properties of resistant glucans were characterized before and after in vitro simulated digestion. The results demonstrated that high-pressure processing (HPP) led to the complete disappearance of crystal peaks and increased the efficiency of the two enzymes (α-amylase and transglucosidase). Moreover, α-1,6 and ß-linkages were abundant in the connecting parts of the long and branching chains in the resulting resistant glucans, thus hindering the ability of digestive enzymes to hydrolyze short chains with a degree of polymerization (DP) ≤ 6. In addition, transglucosidase activity led to a higher proportion of short chains (DP 3-6), further promoting indigestibility. We demonstrated that, without rectification and decolorization, the dietary fiber content was >75 %, and the degree of branching increased to 50.9 %, indicating higher indigestibility than that resistant glucans produced by traditional methods.

Effects of soluble dietary fibers on the viscosity property and digestion kinetics of corn starch digesta.

Four soluble dietary fibers (SDFs) were fortified with corn starch (CS) at different concentrations to match the same viscosity equivalents. The mixtures were subjected to a simulated digestion procedure to study the effects of SDFs on viscosity properties and digestion kinetics of CS. Results showed that SDFs increased the hydration property and decreased the water mobility of digesta. During digestion process, SDFs increased the apparent viscosity of digesta to some extent, and showed significant difference to delay the decay of digesta viscosity (kv). The amylolysis inhibitory ability was similar when each SDF was present at the same viscosity equivalent, however, significant differences were found on the digestion rate constant of k2. Linear correlations between kv and k2 were established for 1 and 2 equivalent groups. These results demonstrated that SDFs could delay the digestion process as chemistry differences, which related to their ability on delaying the change of digesta viscosity.

The Protective Effects and Mechanisms of Apelin/APJ System on Ischemic Stroke: A Promising Therapeutic Target.

The orphan receptor APJ and its endogenous ligand apelin, which are expressed in the brain, are the major components of the apelin/APJ system. Growing evidence shows that the apelin/APJ system plays a vital role in the pathophysiology of cerebral ischemic injury. Targeting the apelin/APJ system may have protective effects on cerebral ischemic injury. In this review, we sum up the latest research progress relating to the actions and therapeutic potential of the apelin/APJ system in ischemic stroke. An in-depth knowledge of the pathophysiological effects of the apelin/APJ system and the underlying mechanisms will help to develop novel therapeutic interventions for ischemic stroke.

Evaluation of Reactivity of Monoclonal Antibodies Against Omp25 of Brucella spp.

Brucellosis is a serious zoonosis occurring mainly in developing countries, and its diagnosis is largely dependent on serologic detection and bacterial culture. In this study, we developed the murine monoclonal antibodies (mAbs) against a conserved and major outer membrane protein 25 (Omp25) of Brucella species (B. spp.) for use in clinical diagnosis. The mAbs to Omp25 were produced by hybridoma technique, which were utilized for developing various immunoassays for detection of Brucellae, including Western blot (WB), enzyme-linked immunosorbent assay (ELISA), immunochemical staining (ICS), immunofluorescence staining (IFS), and flow cytometry assay (FCM). A number of five mAbs (2B10, 4A12, 4F10, 6C12, and 8F3) specific to Omp25 were selected, including 2 IgG1, 2 IgG2a, and 1 IgG2b. Among them, mAbs 6C12, 8F3, and 4A12 reacted highly with B. melitensis (M5-90), B. abortus (S19, 104M, and 2308), and B. suis strain (S2). No cross-reactivity with Yersinia enterocolitica O:9, Salmonella spp., and Escherichia coli was found. By mapping Omp25 epitopes, mAb 6C12 was found as reacting with a semi-conformational epitope, and mAbs 4A12 and 8F3 as recognizing a different linear epitope, respectively. The paired mAbs were tested for detecting Brucella species, suggesting that 8F3 was suitable for solid phase capture and 6C12 or 4A12 was suitable for conjugation with HRP for detection of Brucella Omp25 in ELISA. The FCM was established by mAb 6C12 for detecting intracellular Brucellae-infected peripheral blood mononuclear cells (PBMCs) from brucellosis patients. In conclusion, mAbs against Omp25 are precious reagents for detection of Brucellae in clinical samples with various immunoassays. mAb 6C12-based FCM could be potentially used for the monitoring of therapeutic efficacy for brucellosis in clinical practice.

Interaction between barley ß-glucan and corn starch and its effects on the in vitro digestion of starch.

A ß-glucan was extracted from hull-less barley (HBBG) and its effects on the solution properties and in vitro digestion of corn starch (CS) were studied. Rheological results showed that HBBG diminished the gelling ability and increased the apparent viscosity of CS solution. The critical concentration was lowered from 1.10% (CS) to 0.48% (CS/HBBG mixture), and the slow relaxation component T22 decreased from 1417.47 to 464.16 ms after the incorporation of HBBG to CS solution. In vitro digestion study indicated that HBBG significantly increased the apparent viscosity of digesta and inhibited the starch hydrolysis and glucose diffusion. The entanglement and overlap formed by HBBG and CS interaction and aggregates of HBBG itself were considered to enhance the viscosity, thus limiting the water mobility of the system, reducing the contact of digestive enzyme with starch and diffusion of glucose to the small intestinal microvilli. This study suggests that HBBG can be recognized as an important ingredient in starch food to reduce postprandial glycemic responses.