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1.
Int J Mol Sci ; 24(2)2023 Jan 12.
Artigo em Inglês | MEDLINE | ID: mdl-36674988

RESUMO

Plastic waste is a serious problem in modern society. Every day, mankind produces tons of waste that must be disposed of or recycled. The most common types of plastic waste are disposable tableware, bags, packaging, bottles, and containers, and not all are recycled. Therefore, there is a great interest in producing environmentally friendly disposable materials. In this study, modified gelatin blends using polysaccharides (e.g., agarose, starch) were produced to obtain a stable coating. Various techniques were used to characterize the obtained bioplastics, including FTIR spectroscopy (Fourier-transform infrared spectroscopy), TGA (thermogravimetric analysis)/DSC (differential scanning calorimetry), contact angle measurements, and surface energy characterization. We also investigated the influence of thermal and microbiological degradation on the properties of the biocomposite. The addition of agarose increased the hardness of the blend by 27% compared to the control sample without added polysaccharides. Increases were also observed in the surface energy (24%), softening point (15%), and glass transition temperature (14%) compared to the control sample. The addition of starch to the biopolymer increased the softening point by 15% and the glass transition temperature by 6%. After aging, both blends showed an increase in hardness of 26% and a decrease in tensile strength of 60%.


Assuntos
Gelatina , Amido , Amido/metabolismo , Gelatina/química , Sefarose , Solventes , Polissacarídeos , Plásticos , Espectroscopia de Infravermelho com Transformada de Fourier
2.
Molecules ; 28(2)2023 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-36677869

RESUMO

The large amount of waste generated by the orange juice industry has sparked the interest of many researchers in incorporating recycling systems and following a much more sustainable circular economy model. This work proposes the valorization of the co-product generated in the orange juice extraction industry after freeze-drying for its subsequent reuse as a natural ingredient in the food industry. In addition, the possible protective effect of gum Arabic and corn starch esterified with octenyl succinic groups, in proportions optimised in previous studies 0.25 and 0.45 g/g orange co-product dry solutes, on the main bioactive compounds of orange peel during the freeze-drying process has been studied. The samples were characterised for their content of vitamin C (ascorbic and dehydroascorbic acids), flavonoids (hesperidin and narirutin), total phenols and total carotenoids, as well as their antioxidant capacity (DPPH and FRAP assays). In addition, samples were digested, mimicking the human enzymatic oral gastro-intestinal digestion process, and the bioaccessibility of the bioactive compounds was evaluated. It was observed that the addition of both biopolymers improved the stability of the hydrophilic compounds during freeze-drying. This conservative effect was more remarkable for higher biopolymer concentrations. However, no protective effect on carotenoid compounds was observed. This trend was reflected in the antioxidant activity of the different samples. In addition, the incorporation of biopolymers improved the bioaccessibility of the bioactive compounds studied. In conclusion, the results supported the feasibility of the freeze-dried orange juice co-product as a natural, sustainable source of health-promoting compounds.


Assuntos
Citrus sinensis , Humanos , Citrus sinensis/química , Goma Arábica/metabolismo , Amido/metabolismo , Antioxidantes/química , Ácido Ascórbico/metabolismo , Carotenoides/metabolismo
3.
BMC Microbiol ; 23(1): 32, 2023 01 28.
Artigo em Inglês | MEDLINE | ID: mdl-36707764

RESUMO

BACKGROUND: Interactions between diet, stress and the gut microbiome are of interest as a means to modulate health and performance. Here, in vitro fermentation was used to explore the effects of a sudden change in diet, 21 days sole sustenance on the Meal, Ready-to-Eat (MRE) U.S. military combat ration, on inter-species competition and functional potential of the human gut microbiota. Human fecal samples collected before and after MRE intervention or consuming a habitual diet (HAB) were introduced to nutrient-rich media supplemented with starch for in vitro fermentation under ascending colon conditions. 16S rRNA amplicon and Whole-metagenome sequencing (WMS) were used to measure community composition and functional potential. Specific statistical analyses were implemented to detect changes in relative abundance from taxa, genes and pathways. RESULTS: Differential changes in relative abundance of 11 taxa, Dorea, Lachnospira, Bacteroides fragilis, Akkermansia muciniphila, Bifidobacterium adolescentis, Betaproteobacteria, Enterobacteriaceae, Bacteroides egerthii, Ruminococcus bromii, Prevotella, and Slackia, and nine Carbohydrate-Active Enzymes, specifically GH13_14, over the 24 h fermentation were observed as a function of the diet intervention and correlated to specific taxa of interest. CONCLUSIONS: These findings suggest that consuming MRE for 21 days acutely effects changes in gut microbiota structure in response to carbohydrate but may induce alterations in metabolic capacity. Additionally, these findings demonstrate the potential of starch as a candidate supplemental strategy to functionally modulate specific gut commensals during stress-induced states.


Assuntos
Microbioma Gastrointestinal , Humanos , Microbioma Gastrointestinal/genética , RNA Ribossômico 16S/genética , Dieta , Fezes/microbiologia , Carboidratos , Amido/metabolismo , Suplementos Nutricionais
4.
Int J Mol Sci ; 24(1)2023 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-36614232

RESUMO

Drought is one of the main environmental factors limiting plant growth and development. The AP2/ERF transcription factor (TF) ERF194 play key roles in poplar growth and drought-stress tolerance. However, the physiological mechanism remains to be explored. In this study, the ERF194-overexpression (OX), suppressed-expression (RNA interference, RNAi), and non-transgenic (WT) poplar clone 717 were used to study the physiology role of ERF194 transcription factor in poplar growth and drought tolerance. Morphological and physiological methods were used to systematically analyze the growth status, antioxidant enzyme activity, malondialdehyde (MDA), soluble sugars, starch, and non-structural carbohydrate (NSC) contents of poplar. Results showed that, compared with WT, OX plants had decrease in plant height, internode length, and leaf area and increased number of fine roots under drought stress. In addition, OX had higher water potential, activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD), contents of chlorophyll, soluble sugar, starch, and NSC, implying that ERF194 positively regulates drought tolerance in poplar. The growth status of RNAi was similar to those of WT, but the relative water content and CAT activity of RNAi were lower than those of WT under drought treatment. Based on the transcriptome data, functional annotation and expression pattern analysis of differentially expressed genes were performed and further confirmed by RT-qPCR analysis. Gene ontology (GO) enrichment and gene expression pattern analysis indicated that overexpression of ERF194 upregulated the expression of oxidoreductases and metabolism-related genes such as POD and SOD. Detection of cis-acting elements in the promoters suggested that ERF194 may bind to these genes through MeJA-responsive elements, ABA-responsive elements, or elements involved in defense and stress responses. The above results show that ERF194 improved tolerance to drought stress in poplar by regulating its growth and physiological factors. This study provides a new idea for the role of ERF194 transcription factor in plant growth and drought-stress response.


Assuntos
Fatores de Transcrição , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Antioxidantes/metabolismo , Secas , Peroxidases/metabolismo , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo , Peroxidase/metabolismo , Água/metabolismo , Amido/metabolismo , Estresse Fisiológico/genética , Regulação da Expressão Gênica de Plantas
5.
BMC Med ; 21(1): 27, 2023 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-36691017

RESUMO

BACKGROUND: Salivary amylase, encoded by the AMY1 gene, initiate the digestion of starch. Whether starch intake or AMY1 copy number is related to disease risk is currently rather unknown. The aim was to investigate the association between starch intake and AMY1 copy number and risk of cardiovascular disease (CVD) and mortality and whether there is an interaction. In addition, we aim to identify CVD-related plasma proteins associated with starch intake and AMY1 copy number. METHODS: This prospective cohort study used data from 21,268 participants from the Malmö Diet and Cancer Study. Dietary data were collected through a modified diet history method and incident CVD and mortality were ascertained through registers. AMY1 gene copy number was determined by droplet digital polymerase chain reaction, a risk score of 10 genetic variants in AMY1 was measured, and a total of 88 selected CVD-related proteins were measured. Cox proportional hazards regression was used to analyze the associations of starch intake and AMY1 copy number with disease risk. Linear regression was used to identify plasma proteins associated with starch intake and AMY1 copy number. RESULTS: Over a median of 23 years' follow-up, 4443 individuals developed CVD event and 8125 died. After adjusting for potential confounders, a U-shape association between starch intake and risk of CVD (P-nonlinearity = 0.001) and all-cause mortality (P-nonlinearity = 0.03) was observed. No significant association was found between AMY1 copy number and risk of CVD and mortality, and there were no interactions between starch intake and AMY1 copy number (P interaction > 0.23). Among the 88 plasma proteins, adrenomedullin, interleukin-1 receptor antagonist protein, fatty acid-binding protein, leptin, and C-C motif chemokine 20 were associated with starch intake after adjusting for multiple testing. CONCLUSIONS: In this large prospective study among Swedish adults, a U-shaped association between starch intake and risk of CVD and all-cause mortality was found. Several plasma proteins were identified which might provide information on potential pathways for such association. AMY1 copy number was not associated with CVD risk or any of the plasma proteins, and there was no interaction between starch intake and AMY1 copy number on disease risk.


Assuntos
Doenças Cardiovasculares , alfa-Amilases Salivares , Humanos , Variações do Número de Cópias de DNA , Amido/metabolismo , Estudos Prospectivos , Amilases/genética , alfa-Amilases Salivares/genética , alfa-Amilases Salivares/metabolismo , Dosagem de Genes , Proteínas Sanguíneas/genética
6.
Appl Microbiol Biotechnol ; 107(2-3): 971-983, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36622426

RESUMO

Microalgae are promising feedstock for renewable fuels. The accumulation of oils in microalgae can be enhanced by nanoparticle exposure. However, the nanoparticles employed in previous studies are mostly non-biodegradable, which hinders nanoparticles developing as promising approach for biofuel production. We recently reported the engineered resin nanoparticles (iBCA-NPs), which were found to be biodegradable in this study. When the cells of green microalga Chlamydomonas reinhardtii were exposed to the iBCA-NPs for 1 h, the cellular triacyclglycerols (TAG) and starch contents increased by 520% and 60% than that in the control. The TAG production improved by 1.8-fold compared to the control without compromised starch production. Additionally, the content of total fatty acids increased by 1.3-fold than that in control. Furthermore, we found that the iBCA-NPs addition resulted in increased cellular reactive oxygen species (ROS) content and upregulated the activities of antioxidant enzymes. The relative expressions of the key genes involved in TAG and starch biosynthesis were also upregulated. Overall, our results showed that short exposure of the iBCA-NPs dramatically enhances TAG and starch accumulation in Chlamydomonas, which probably resulted from prompt upregulated expression of the key genes in lipid and starch metabolic pathways that were triggered by over-accumulated ROS. This study reported a useful approach to enhance energy-rich reserve accumulation in microalgae. KEY POINTS: 1. The first attempt to increase oil and starch in microalgae by biodegradable NPs. 2. The biodegradability of iBCA-NPs by the BOD test was more than 50% after 28 days. 3. The iBCA-NPs induce more energy reserves than that of previously reported NPs.


Assuntos
Chlamydomonas reinhardtii , Chlamydomonas , Microalgas , Nanopartículas , Chlamydomonas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Triglicerídeos/metabolismo , Chlamydomonas reinhardtii/genética , Chlamydomonas reinhardtii/metabolismo , Amido/metabolismo , Microalgas/metabolismo
7.
J Agric Food Chem ; 71(2): 1056-1066, 2023 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-36595531

RESUMO

Plant fructokinase (FRK) guarantees the growth and development of higher plants by participating in carbohydrate metabolism. In this study, a new fructokinase, OsFRK3, was identified using bioinformatics analysis, enzyme assay, bacterial growth assay, and yeast complementation test. Then, we created OsFRK3 knockout transgenic lines (osfrk3-1 and osfrk3-2) by the CRISPR/Cas9 technology. We found that the 1000-grain weight decreased notably (approximately -3.6% and -6.1%, respectively) in osfrk3-1 and osfrk3-2. Evidently decreased grain width, grain thickness, and endosperm filling rate were detected in the osfrk3 mutants (osfrk3-1 and osfrk3-2) compared with those of the WT. In addition, the content of seed total starch was significantly decreased by 3.42 and 4.80% in osfrk3 lines, compared with that in the WT. The level of maltose was significantly reduced in the mutants, while that of sucrose and fructose was obviously increased in the mutants. The transcript levels of OsGBSS1, OsBEIIb, OsPFP1ß, and OsAGPL1 were significantly decreased in the osfrk3 mutants. These results suggest that OsFRK3 may positively regulate the accumulation of starch through influencing the sugar metabolism.


Assuntos
Oryza , Amido , Amido/metabolismo , Endosperma/genética , Endosperma/metabolismo , Grão Comestível/metabolismo , Sementes/genética , Sementes/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regulação da Expressão Gênica de Plantas
8.
Appl Microbiol Biotechnol ; 107(2-3): 491-515, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36512032

RESUMO

Since the role of intestinal microbiota in metabolism was understood, the importance of dietary components such as fibres and prebiotics, which affect the modulation of microbiota, has been increasing day by day. While all prebiotic components are considered dietary fibre, not every dietary fibre is considered a prebiotic. While fructooligosaccharides, galactooligosaccharides, inulin, and galactans are considered prebiotics, other fermentable carbohydrates are considered candidate prebiotic components based on in vitro and preclinical studies. Resistant starch, one of such carbohydrates, is considered a potential prebiotic component when it is made resistant to digestion naturally or chemically. In this review, both in vitro and in vivo studies in which the prebiotic capacity of type II, type III, and type IV resistant starch isolated from food and produced commercially was assessed were analyzed. According to the results of current studies, certain types of resistant starch are thought to have a high prebiotic capacity, and they may be candidate prebiotic components although positive results have not been achieved in all studies. KEY POINTS: • Resistant starch is undigested in the small intestine and is fermented in the large intestine. • Resistant starch fermentation positively affects the growth of Bifidobacterium and Lactobacillus. • Resistant starch can be considered a prebiotic ingredient.


Assuntos
Prebióticos , Amido Resistente , Amido Resistente/metabolismo , Amido/metabolismo , Fibras na Dieta/metabolismo , Inulina/metabolismo , Fermentação
9.
Environ Res ; 216(Pt 3): 114722, 2023 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-36343710

RESUMO

This study elucidated the changes in the short-term response to organic load shocks of the anaerobic digestion (AD) microbiome acclimatized to a simple substrate and a complex substrate. Batch vial reactors were inoculated with AD sludge acclimatized to either a simple (starch and hipolypeptone) or a complex (dog food and starch) substrate, both with carbon-to-nitrogen ratio of 25. Organic loads in the form of an easily degradable substrate mix (starch and hipolypeptone) with concentrations varying from 0 to 5 g VS/L were applied to the reactors. Runs utilizing the inoculum acclimatized to a complex substrate sustained its methane productivity despite the high organic load shocks which the inoculum acclimatized to a simple substrate was unable to handle efficiently. The alpha-diversity of the microbiome decreased with increase in organic load for inoculum acclimatized with a simple substrate but was unaffected for the case of the inoculum acclimatized with a complex substrate. LactobacillalesandCloacimonadales were inferred to be major players in starch degradation pathways for the inoculum acclimatized using a simple substrate as predicted by the bioinformatics package PICRUSt2. However, acclimatizing using a complex substrate did not support their growth and were replaced by Coriobacteriales which provided higher flexibility in terms of the predicted regulated metabolic functions. The predicted functional regulation of Synergistales and Syntrophales increased with acclimatization using a complex substrate which also showed increase in the flexibility of the microbiome towards handling organic load shocks. Acetoclastic pathway was upregulated with increase in organic load regardless of the acclimatization substrate while the hydrogenotrophic pathway was downregulated. Overall, acclimatization using a complex substrate increased the robustness and flexibility of the microbiome towards organic load shocks.


Assuntos
Reatores Biológicos , Microbiota , Animais , Cães , Anaerobiose , Metano , Esgotos , Aclimatação , Amido/metabolismo
10.
J Dairy Sci ; 106(2): 1002-1012, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36543642

RESUMO

The objective of this study was to determine the effects of including exogenous amylolytic or fibrolytic enzymes in a diet for high-producing dairy cows on in vitro ruminal fermentation. Eight dual-flow continuous-culture fermentors were used in a replicated 4 × 4 Latin square. The treatments were control (CON), a xylanase and glucanase mixture (T1), an α-amylase mixture (T2), or a xylanase, glucanase, and α-amylase mixture (T3). Treatments were included at a rate of 0.008% of diet dry matter (DM) for T1 and T2 and at 0.02% for T3. All treatments replaced the equivalent amount of soybean meal in the diet compared with CON. All diets were balanced to have the same nutrient composition [30.2% neutral detergent fiber (NDF), 16.1% crude protein (CP), and 30% starch; DM basis], and fermentors were fed 106 g/d divided into 2 feedings. At each feeding, T2 was pipetted into the respective fermentor and an equivalent amount of deionized water was added to each fermentor to eliminate potential variation. Experimental periods were 10 d (7 d for adaptation and 3 d for sample collection). Composite samples of daily effluent were collected and analyzed for volatile fatty acids (VFA), NH3-N, and lactate concentrations, degradability of DM, organic matter, NDF, CP, and starch, and flow and metabolism of N. Samples of fermentor contents were collected from each fermentor at 0, 1, 2, 4, 6, and 8 h after feeding to determine kinetics of pH, NH3-N, lactate, and VFA concentrations over time. All data were analyzed using PROC GLIMMIX of SAS (SAS Institute Inc.), and the repeated variable of time was included for kinetics measurements. Treatment did not affect mean pH, degradability, N flow and metabolism, or the concentrations of VFA, NH3-N, or lactate in the effluent samples. Treatment did not affect pH, acetate:propionate ratio, or the concentrations of lactate, NH3-N, total VFA, acetate, propionate, butyrate, isobutyrate, valerate, or caproate. However, the concentration of total VFA tended to change at each time point depending upon the treatment, and T2 tended to have a greater proportion of 2-methylbutyrate and isovalerate than CON, T1, or T3. As 2-methylbutyrate and isovalerate are branched-chain VFA that are synthesized from branched-chain amino acids, T2 may have an increased fermentation of branched-chain amino acids or decreased uptake by fibrolytic microorganisms. Although we did not observe changes in N metabolism due to the enzymes, there could be changes in microbial populations that utilize branched-chain VFA. Overall, the tested enzymes did not improve in vitro ruminal fermentation in the diet of high-producing dairy cows.


Assuntos
Lactação , Propionatos , Animais , Bovinos , Feminino , alfa-Amilases/metabolismo , Ração Animal/análise , Dieta/veterinária , Digestão , Ácidos Graxos Voláteis/metabolismo , Fermentação , Lactatos/metabolismo , Propionatos/metabolismo , Rúmen/metabolismo , Amido/metabolismo
11.
J Agric Food Chem ; 71(1): 615-625, 2023 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-36537359

RESUMO

Optimized source-sink interactions are determinants of both rice yield and quality. However, most source genes have not been well studied in rice, a major grain crop. In this study, OsBMY4 and OsISA3, the key ß-amylase and debranching enzymes that control transient starch degradation in rice leaves, were co-overexpressed in rice in order to accelerate starch degradation efficiency and increase the sugar supply for sink organs. Systematic analyses of the transgenic rice indicated that co-overexpression of OsBMY4 and OsISA3 not only promoted rice yield and quality, but also improved seed germination and stress tolerance. Moreover, since the OsBMY4 gene has not been characterized, we generated osbmy4 mutants using CRIPSR/Cas9 gene editing, which helped to reveal the roles of ß-amylase in rice yield and quality. This study demonstrated that specific modulation of the expression of some key source genes improves the source-sink balance and leads to improvements in multiple key traits of rice seeds.


Assuntos
Oryza , beta-Amilase , Oryza/genética , Oryza/metabolismo , beta-Amilase/genética , Sementes/genética , Sementes/metabolismo , Grão Comestível/metabolismo , Amido/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
12.
J Plant Physiol ; 280: 153902, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36565529

RESUMO

Plants are regularly challenged by unfavorable environmental conditions. As climate change continues, adverse situations such as drought, heat, and cold are expected to increase and become more severe. Most starchy crops are affected by such stresses. In recent years, researchers have made many new discoveries about starch metabolism in general and also on granule structure, including effects on starch following longer-term temperature stresses. However, in this study, we focus on short-term temperature stress on storage starch granule properties. Here our knowledge is less and it is likely that also short-term temperature stresses can affect various starch parameters. Therefore, we see a need for this type of analysis and discuss the matter in more detail and we conclude that a deeper knowledge particularly of starch granule parameters could allow targeted breeding of cultivars that exhibit different starch characteristics as a result of short-term stress. For these reasons, we are convinced that more comprehensive research on the effects of short-term temperature stress on starch granule characteristics is important, necessary, and timely.


Assuntos
Temperatura Alta , Amido , Amido/metabolismo , Temperatura , Temperatura Baixa , Produtos Agrícolas/metabolismo
13.
Ecotoxicol Environ Saf ; 249: 114379, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36508814

RESUMO

High-molecular-weight PAHs (HMW-PAHs) in soil cannot be easily degraded. However, nutrient supplementation could stimulate the growth of exogenously added strains to enhance the degradation of HMW-PAHs in polluted soil. This study evaluated the applicability of Fusarium sp. ZH-H2, a polycyclic aromatic hydrocarbon (PAH)-degrading strain isolated by our research group, for the bioremediation of contaminated soil from the Hebei coal mining area in China. A soil incubation experiment was conducted to investigate the effect of two carbon sources and different carbon, nitrogen, and phosphorus (C:N:P) ratios on the remediation of high-molecular-weight PAHs (HMW-PAHs) in soil by Fusarium sp. ZH-H2, as well as the induction of lignin peroxidase activity. Our findings indicated that the HDF2 treatment (equal parts of humic acid and starch as carbon sources at a 50:1:0.5 C:N:P ratio) enhanced the removal rate of total HMW-PAHs from soil, reaching a maximum removal rate of 37.15 %. The removal rates of Pyr (a 4-ring PAH), BaP (a 5-ring PAH), and BghiP (a 6-ring PAH) were the highest in HDF2 treatment, and the removal rates were 39.51 %, 54.63 %, and 38.60 %, respectively. Compared with the ZH-H2 treatment, different carbon sources and C:N:P ratios significantly induced soil lignin peroxidase activity and the HDF2 treatment also resulted in the highest enzyme activity (up to 34.68 U/L). Furthermore, there was a significant or highly significant linear positive correlation between the removal rate of HMW-PAHs and enzyme activity in all cases. Our findings suggest that the optimal HMW-PAH degradation performance and enhancement of lignin peroxidase activity by ZH-H2 were achieved when both starch and humic acid were used as carbon sources at a C:N:P ratio of 50:1:0.5.


Assuntos
Fusarium , Hidrocarbonetos Policíclicos Aromáticos , Poluentes do Solo , Solo , Fusarium/metabolismo , Substâncias Húmicas , Hidrocarbonetos Policíclicos Aromáticos/análise , Carbono/metabolismo , Poluentes do Solo/análise , Biodegradação Ambiental , Amido/metabolismo , Microbiologia do Solo
14.
J Dairy Sci ; 106(2): 1453-1463, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36526457

RESUMO

Intestinal hyperpermeability and subsequent immune activation alters nutrient partitioning and thus, decreases productivity. Developing experimental models of intestinal barrier dysfunction in heathy cows is a prerequisite in identifying nutritional strategies to mitigate it. Six cannulated Holstein cows (mean ± standard deviation, 37 ± 10 kg/d milk yield; 219 ± 97 d in milk; 691 ± 70 kg body weight) were used in a replicated 3 × 3 Latin square design experiment with 21-d periods (16-d wash-out and 5-d challenge) to evaluate either feed restriction or hindgut acidosis as potential models for inducing intestinal hyperpermeability. Cows were randomly assigned to treatment sequence within square and treatment sequences were balanced for carryover effects. Treatments during the challenge were (1) control (CTR; ad libitum feeding); (2) feed restriction (FR; total mixed ration fed at 50% of ad libitum feed intake); and (3) resistant starch (RS; 500 g of resistant starch infused in abomasum once a day as a pulse-dose 30 min before morning feeding). The RS (ActiStar RT 75330, Cargill Inc.) was tapioca starch that was expected to be resistant to enzymatic digestion in the small intestine and highly fermentable in the hindgut. Blood samples were collected 4 h after feeding on d 13 and 14 of the wash-out periods (baseline data used as covariate), and on d 1, 3, and 5 of the challenge periods. Fecal samples were collected 4 and 8 h after the morning feeding on d 14 of the wash-out periods and d 5 of the challenge periods. By design, FR decreased dry matter intake (48%) relative to CTR and RS, and this resulted in marked reductions in milk and 3.5% FCM yields over time, with the most pronounced decrease occurring on d 5 of the challenge (34 and 27%, respectively). Further, FR increased somatic cell count by 115% on d 5 of the challenge relative to CTR and RS. Overall, FR increased nonesterified fatty acids (159 vs. 79 mEq/L) and decreased BHB (8.5 vs. 11.2 mg/dL), but did not change circulating glucose relative to CTR. However, RS had no effect on production or metabolism metrics. Resistant starch decreased fecal pH 8 h after the morning feeding (6.26 vs. 6.81) relative to CTR and FR. Further, RS increased circulating lipopolysaccharide binding protein (4.26 vs. 2.74 µg/mL) compared with FR only on d 1 of the challenge. Resistant starch also increased Hp (1.52 vs. 0.48 µg/mL) compared with CTR, but only on d 5 of the challenge. However, neither RS or FR affected concentrations of serum amyloid A, IL1ß, or circulating endotoxin compared with CTR. The lack of consistent responses in inflammatory biomarkers suggests that FR and RS did not meaningfully affect intestinal barrier function. Thus, future research evaluating the effects of hindgut acidosis and FR using more intense insults and direct metrics of intestinal barrier function is warranted.


Assuntos
Lactação , Amido Resistente , Feminino , Bovinos , Animais , Amido Resistente/metabolismo , Amido Resistente/farmacologia , Dieta/veterinária , Abomaso/metabolismo , Leite/metabolismo , Ração Animal/análise , Rúmen/metabolismo , Amido/metabolismo
15.
Enzyme Microb Technol ; 164: 110176, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36529061

RESUMO

Bifunctional debranching-enzyme amylopullulanases belong to the glycoside hydrolases (GHs) family and catalyze both the hydrolysis of α-1,4 and α-1,6 glycosidic bonds in starch, pullulan, amylopectin and glycogen polysaccharides. Among these, especially thermostable ones are essential in starch processing applications. In this study, we focused to elucidate the complete sequence of the apu gene and the role of C-term domains on biochemical properties and enzyme activity of Thermoanaerobacter brockii brockii amylopullulanase (TbbApu). After the gene sequence was defined, C- term truncated variants were constructed. The most suitable host organism and expression vector were determined as E. coli BL21(DE3) and pET-28a(+) depending on the highest yield/biomass ratio for recombinant production of all constructs. It was seen that the expression yield increased approximately threefold in the case of the SH3 region truncation. In the biochemical characterization, TbbApu and its truncated variants exhibited maximum activity at 70 °C and 75 °C for pullulan and starch hydrolysis respectively, and the optimum pH of TbbApu were 6.5 and 6 for truncated variants. Moreover, hydrolysis activities of all recombinant enzymes were enhanced by Mn2+, Co2+ and Cu2+, detergents, and almost all organic solvents; except butanol, DMF and DMSO. All recombinant amylopullulanases remained 80% stable up to 80 °C in the wide range of pH and also retained > 85% stability in the presence of defined volatile organic solvents. No significant difference was observed between the raw starch adsorption capacity and the specific activity of the three variants. These results indicated that the C-terminal regions of TbbApu are non-essential for the enzyme activity, stability and substrate binding capacity; furthermore, hexane and acetone organic solvents enhanced both pullulanase and α-amylase activity of these enzymes, interestingly. With these features, TbbApu and its truncated variants are distinguished from other thermophilic amylopullulanases and also make them promising candidates for industrial use.


Assuntos
Proteínas de Bactérias , Escherichia coli , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Bactérias/metabolismo , Amido/metabolismo , Glicosídeo Hidrolases/metabolismo , Solventes , Clonagem Molecular , Concentração de Íons de Hidrogênio , Estabilidade Enzimática , Especificidade por Substrato
16.
Phytochemistry ; 207: 113563, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36528118

RESUMO

Hydrogen stable isotope analyses (δ2H) of plant derived organic compounds are a useful tool for ecological, environmental, and palaeoclimatological research. However, during organic compound synthesis, variable biosynthetic 2H-fractionation has been suggested to occur as a result of changes in plant carbon fluxes. So far, inference has been based on examining the δ2H patterns of plant compounds along environmental gradients, among plant species, and between plant organs. In an alternative approach, we used four plant species with four different types of mutations that cause impaired starch synthesis to assess whether variability in carbon metabolism affects the biosynthetic 2H-fractionation during cellulose, phytol, and acetogenic lipid synthesis. We found that mutants with impaired starch synthesis always had higher cellulose and phytol δ2H values compared to the wild type. By contrast, 2H-fractionation during acetogenic lipid biosynthesis generally did not show strong metabolic sensitivity. We rationalise these differences by considering the biosynthetic pathway of each compound and the likely source of the variable isotope fractionation. In different organic compounds, the sensitivity of variable biosynthetic 2H-fractionation to changes in C-metabolism depends on incorporation of specific H atoms from precursor molecules. As such, we determined that the similar increase in cellulose and phytol δ2H values as an effect of impaired starch synthesis most likely originates in triose-phosphates.


Assuntos
Carbono , Hidrogênio , Hidrogênio/metabolismo , Isótopos , Plantas/metabolismo , Compostos Orgânicos/metabolismo , Celulose/metabolismo , Lipídeos , Amido/metabolismo , Fitol/metabolismo , Isótopos de Carbono/metabolismo , Folhas de Planta/metabolismo
17.
Poult Sci ; 102(2): 102362, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36566658

RESUMO

This experiment investigated the effects of different dietary starch sources on the growth and glucose metabolism of geese. A total of 240 healthy 35-day-old male geese were selected and randomly divided into 4 groups, with 6 replicates per group and 10 geese per replicate. Four types of diets were prepared, with glutinous rice (rapidly-digestible starch), corn, indica rice and high amylose as their starch sources, and fed for 28 d. Results showed that after consuming different feeds, the blood glucose of geese first increased and then decreased, reaching its maximum value 0.5 h after feeding, and there were significant differences between the groups (P < 0.05). The body weight of the corn and indica rice group geese at 63 d was higher than that of the high amylose group (P < 0.05). The serum total cholesterol (TCHO) content in the glutinous rice and corn groups was higher than in the high amylose group (P < 0.05). The serum insulin content in the glutinous rice group was lower than in the corn and high amylose groups (P < 0.05), while the glucagon content was higher (P < 0.05). The α-amylase activities of the pancreas, jejunal chyme, and jejunal mucosa in the glutinous rice group were higher than in the indica rice and high amylose groups (P < 0.05). The liver glycogen content in the glutinous rice group was higher than the other groups (P < 0.05). The liver glucose-6-phosphate dehydrogenase (G-6-PD) content in the glutinous rice group was higher than the high amylose group's (P < 0.05), but the glycogen synthase kinase-3 ß (GSK-3ß) content was lower (P < 0.05). In conclusion, the corn and indica rice diets had a positive effect on the growth performance of the geese, while the high amylose diet had a negative effect. The glutinous rice diet leads to rapid release of glucose, strengthening glucose metabolism pathways such as glycogen synthesis and the pentose phosphate pathway, and further influencing lipid metabolism.


Assuntos
Amilose , Oryza , Masculino , Animais , Amilose/metabolismo , Amilose/farmacologia , Gansos/metabolismo , Glicogênio Sintase Quinase 3 beta , Galinhas/metabolismo , Amido/metabolismo , Carboidratos da Dieta , Glucose
18.
Plant Physiol Biochem ; 194: 394-405, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36481708

RESUMO

The sugar transporter SWEET plays a role in plant growth, carbon allocation, and abiotic stress resistance. We examined the function of SWEET in cassava (Manihot esculenta Crantz) under water and salt stress. Bioinformatics, subcellular localization, yeast deficient complementation, and virus-induced gene silencing (VIGS) were used to examine the function of SWEET in cassava. Twenty-eight MeSWEETs genes were found based on the conserved domain MtN3/saliva of SWEET transporters, two MeSWEET15a/b of them were identified by phylogenetic analysis, which were located on the cell membrane. They transfer sucrose, fructose, glucose, and mannitol from culture media to yeast cells, predominately transferring sucrose via bleeding fluid saps in plant. Leaf sucrose content was increased in MeSWEET15a/b-silenced cassava plants, resulting in changes in carbon distribution, with an increase in starch accumulation in the leaves and a decrease in starch accumulation in the roots. The silencing of MeSWEET15a/b genes led to tolerance to water and salt stress, consistent with a high accumulation of osmolytes, and low lipid membrane peroxidation. Changes in sugar distribution increased the expression of MeTOR and MeE2Fa in pTRV2-MeSWEET15a and pTRV2-MeSWEET15b cassava leaves. MeSWEET15a/b acts as pivotal modulators of sugar distribution and tolerance to water and high salt stress in cassava.


Assuntos
Manihot , Água , Água/metabolismo , Açúcares/metabolismo , Manihot/genética , Manihot/metabolismo , Filogenia , Saccharomyces cerevisiae/metabolismo , Amido/metabolismo , Estresse Salino , Sacarose/metabolismo
19.
Waste Manag ; 157: 256-268, 2023 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-36577277

RESUMO

Due to the rising demand for green energy, bioethanol has attracted increasing attention from academia and industry. Limited by the bottleneck of bioethanol yield in traditional corn starch dry milling processes, an increasing number of studies focus on fully utilizing all corn ingredients, especially kernel fiber, to further improve the bioethanol yield. This mini-review addresses the technological challenges and opportunities on the way to achieving the efficient conversion of corn fiber. Significant advances during the review period include the detailed characterization of different forms of corn kernel fiber and the development of off-line and in-situ conversion strategies. Lessons from cellulosic ethanol technologies offer new ways to utilize corn fiber in traditional processes. However, the commercialization of corn kernel fiber conversion may be hampered by enzyme cost, conversion efficiency, and overall process economics. Thus, future studies should address these technical limitations.


Assuntos
Amido , Zea mays , Zea mays/metabolismo , Amido/metabolismo , Etanol , Tecnologia , Fermentação
20.
Plant Mol Biol ; 108(4-5): 363-378, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-34546521

RESUMO

Ostreococcus tauri is a picoalga that contains a small and compact genome, which resembles that of higher plants in the multiplicity of enzymes involved in starch synthesis (ADP-glucose pyrophosphorylase, ADPGlc PPase; granule bound starch synthase, GBSS; starch synthases, SSI, SSII, SSIII; and starch branching enzyme, SBE, between others), except starch synthase IV (SSIV). Although its genome is fully sequenced, there are still many genes and proteins to which no function was assigned. Here, we identify the OT_ostta06g01880 gene that encodes CBM20CP, a plastidial protein which contains a central carbohydrate binding domain of the CBM20 family, and a coiled coil domain at the C-terminus that lacks catalytic activity. We demonstrate that CBM20CP has the ability to bind starch, amylose and amylopectin with different affinities. Furthermore, this protein interacts with OsttaSSIII-B, increasing its binding to starch granules, its catalytic efficiency and promoting granule growth. The results allow us to postulate a functional role for CBM20CP in starch metabolism in green algae. KEY MESSAGE: CBM20CP, a plastidial protein that has a modular structure but lacks catalytic activity, regulates the synthesis of starch in Ostreococcus tauri.


Assuntos
Proteínas de Algas/metabolismo , Clorófitas/metabolismo , Amido/metabolismo , Proteínas de Algas/genética , Sequência de Aminoácidos , Amilopectina/metabolismo , Amilose/metabolismo , Clorófitas/enzimologia , Clorófitas/genética , Clonagem Molecular , Plastídeos , Ligação Proteica , Alinhamento de Sequência
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