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1.
J Appl Microbiol ; 135(6)2024 Jun 03.
Artículo en Inglés | MEDLINE | ID: mdl-38830792

RESUMEN

AIMS: Cytidine, as an important commercial precursor in the chemical synthesis of antiviral and antitumor drugs, is in great demand in the market. Therefore, the purpose of this study is to build a microbial cell factory with high cytidine production. METHODS AND RESULTS: A mutant E. coli NXBG-11-F34 with high tolerance to uridine monophosphate structural analogs and good genetic stability was obtained by atmospheric room temperature plasma (ARTP) mutagenesis combined with high-throughput screening. Then, the udk and rihA genes involved in cytidine catabolism were knocked out by CRISPR/Cas9 gene editing technology, and the recombinant strain E. coli NXBG-13 was constructed. The titer, yield, and productivity of cytidine fermented in a 5 l bioreactor were 15.7 g l-1, 0.164 g g-1, and 0.327 g l-1 h-1, respectively. Transcriptome analysis of the original strain and the recombinant strain E. coli NXBG-13 showed that the gene expression profiles of the two strains changed significantly, and the cytidine de novo pathway gene of the recombinant strain was up-regulated significantly. CONCLUSIONS: ARTP mutagenesis combined with metabolic engineering is an effective method to construct cytidine-producing strains.


Asunto(s)
Citidina , Escherichia coli , Ingeniería Metabólica , Mutagénesis , Escherichia coli/genética , Escherichia coli/metabolismo , Citidina/genética , Citidina/metabolismo , Gases em Plasma , Reactores Biológicos , Edición Génica/métodos , Sistemas CRISPR-Cas , Fermentación , Temperatura
2.
Physiol Mol Biol Plants ; 30(6): 909-919, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38974351

RESUMEN

Bud mutation is a common technique for plant breeding and can provide a large number of breeding materials. Through traditional breeding methods, we obtained a plum plant with bud mutations (named "By") from an original plum variety (named "B"). The ripening period of "By" fruit was longer than that of "B" fruit, and its taste was better. In order to understand the characteristics of these plum varieties, we used transcriptome analysis and compared the gene expression patterns in fruits from the two cultivars. Subsequently, we identified the biological processes regulated by the differentially expressed genes (DEGs). Gene ontology (GO) analysis revealed that these DEGs were highly enriched for "single-organism cellular process" and "transferase activity". KEGG analysis demonstrated that the main pathways affected by the bud mutations were plant hormone signal transduction, starch and sucrose metabolism. The IAA, CKX, ARF, and SnRK2 genes were identified as the key regulators of plant hormone signal transduction. Meanwhile, TPP, the beta-glucosidase (EC3.2.1.21) gene, and UGT72E were identified as candidate DEGs affecting secondary metabolite synthesis. The transcriptome sequencing (RNA-seq) data were also validated using RT-qPCR experiments. The transcriptome analysis demonstrated that plant hormones play a significant role in extending the maturity period of plum fruit, with IAA, CKX, ARF, and SnRK2 serving as the key regulators of this process. Further, TPP, beta-glucosidase (EC3.2.1.21), and UGT72E appeared to mediate the synthesis of various soluble secondary metabolites, contributing to the aroma of plum fruits. The expression of BAG6 was upregulated in "B" as the fruit matured, but it was downregulated in "By". This indicated that "B" may have stronger resistance, especially fungal resistance. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-024-01472-3.

3.
Mol Biol Rep ; 50(10): 7995-8003, 2023 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-37540452

RESUMEN

BACKGROUND: Apricot fruit has great economic value. In the process of apricot breeding using traditional breeding methods, we obtained a larger seedling (named Us) from the original variety (named U). And Us fruit is larger than U, taste better. Therefore, revealing its mechanism is very important for Apricot breeding. METHODS: In this study, de novo assembly and transcriptome sequencing (RNA-Seq) was used to screen the differently expressed genes (DEGs) between U and Us at three development stages, including young fruits stage, mid-ripening stage and mature fruit stage. RESULTS: The results showed that there were 6,753 DEGs at different sampling time. "Cellulose synthase (UDP-forming) activity" and "cellulose synthase activity" were the key GO terms enriched in GO, of which CESA and CSL family played a key role. "Photosynthesis-antenna proteins" and "Plant hormone signal transduction" were the candidate pathways and lhca, lhcb, Aux/IAA and SAUR were the main regulators. CONCLUSION: The auxin signaling pathway was active in Us, of which Aux/IAAs and SAUR were the key fruit size regulators. The low level of lhca and lhcb in Us could reveal the low demand for exogenous carbon, but they increased at mature stage, which might be due to the role of aux, who was keeping the fruit growing. Aux and photosynthesis maight be the main causes of appearance formation of Us fruits. Interestingly, the higher expression of CESA and CSL proved that Us entered the hardening process earlier than U. The advanced developmental progress might also be due to the role of Aux.


Asunto(s)
Frutas , Prunus armeniaca , Frutas/metabolismo , Prunus armeniaca/genética , Plantones/genética , Plantones/metabolismo , Fitomejoramiento , Perfilación de la Expresión Génica , Reguladores del Crecimiento de las Plantas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regulación de la Expresión Génica de las Plantas/genética , Transcriptoma/genética , Ácidos Indolacéticos/metabolismo
4.
Molecules ; 27(16)2022 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-36014425

RESUMEN

Fruits of six varieties of young citrus cultivated in China were collected for phytochemical composition analysis and antioxidant activity determination. The phenolic acids, synephrine, flavone, and flavanone were analyzed using HPLC, and the total phenolic content and antioxidant capacity were determined by Folin-Ciocalteu, Ferric ion reducing antioxidant power (FRAP), 2,2- 1,1-diphenyl-2-picrylhydrazyl (DPPH), and 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) analysis. The results indicated that Ougan variety had the highest total phenolic content (125.18 GAE mg/g DW), followed by the Huyou variety (107.33 mg/g DW), while Wanshuwenzhoumigan variety had the lowest (35.91 mg/g DW). Ferulic acid was the most dominant soluble phenolic acid in the selected young citrus, followed by p-coumaric acid and p-hydroxybenzoic acid, whereas nobiletin and tangeretin were the most abundant flavones in the Ponkan, Ougan, and Wanshuwenzhoumigan varieties. Antioxidant capacity that measured by ABTS, FRAP, and DPPH showed similar trends and was positively correlated with the total phenolic and total flavonoid contents (p < 0.05). Considering the high content of phenolics in the young fruits of Ougan and Huyou variety, those two varieties might be potential resources for extracting phytochemicals for health promotion.


Asunto(s)
Antioxidantes , Citrus , Antioxidantes/química , Citrus/química , Flavonoides/química , Fenoles/química , Fitoquímicos/análisis
5.
Molecules ; 27(20)2022 Oct 17.
Artículo en Inglés | MEDLINE | ID: mdl-36296554

RESUMEN

The emulsification of ultrasonic-assisted prepared octenyl succinic anhydride (OSA) rice starch on curcumin was investigated in the present study. The results indicated that the encapsulation efficiency of curcumin in emulsions stabilized by OSA-ultrasonic treatment rice starch was improved, from 81.65 ± 0.14% to 89.03 ± 0.09%. During the in vitro oral digestion, the particle size and Zeta potential of the curcumin emulsion did not change significantly (p > 0.05). During the in vitro digestive stage of the stomach and small intestine, the particle size of the curcumin emulsion continued to increase, and the absolute potential continued to decrease. Our work showed that OSA-pre-treatment ultrasonic rice starch could improve curcumin bioavailability by increasing the encapsulation efficiency with stronger stability to avoid the attack of enzymes and high intensity ion, providing a way to develop new emulsion-based delivery systems for bioactive lipophilic compounds using OSA starch.


Asunto(s)
Curcumina , Oryza , Emulsiones , Almidón , Anhídridos Succínicos , Ultrasonido
6.
Biotechnol Lett ; 43(6): 1211-1219, 2021 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-33646457

RESUMEN

Cytidine is an important raw material for nucleic acid health food and genetic engineering research. In recent years, it has shown irreplaceable effects in anti-virus, anti-tumor, and AIDS drugs. Its biosynthetic pathway is complex and highly regulated. In this study, overexpression of uracil permease and a nucleoside transporter from Bacillus amyloliquefaciens related to cell membrane transport in Escherichia coli strain BG-08 was found to increase cytidine production in shake flask cultivation by 1.3-fold (0.91 ± 0.03 g/L) and 1.8-fold (1.26 ± 0.03 g/L) relative to that of the original strain (0.70 ± 0.03 g/L), respectively. Co-overexpression of uracil permease and a nucleoside transporter further increased cytidine yield by 2.7-fold (1.59 ± 0.05 g/L) compared with that of the original strain. These results indicate that the overexpressed uracil permease and nucleoside transporter can promote the accumulation of cytidine, and the two proteins play a synergistic role in the secretion of cytidine in Escherichia coli.


Asunto(s)
Bacillus amyloliquefaciens/metabolismo , Citidina/metabolismo , Escherichia coli/crecimiento & desarrollo , Proteínas de Transporte de Nucleósidos/metabolismo , Bacillus amyloliquefaciens/genética , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Técnicas Bacteriológicas/instrumentación , Técnicas de Cultivo Celular por Lotes , Vías Biosintéticas , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Transporte de Membrana/genética , Proteínas de Transporte de Membrana/metabolismo , Proteínas de Transporte de Nucleósidos/genética , Ingeniería de Proteínas
7.
Molecules ; 25(23)2020 Nov 30.
Artículo en Inglés | MEDLINE | ID: mdl-33266001

RESUMEN

In order to broaden the application of potato pulp pectic polysaccharide (PPP) in stabilizing acidified milk drinks (AMDs) and investigate the stabilizing effect and physical properties of AMDs prepared with PPP, a comparative study was made among PPP, commercial high methoxyl pectin (HMP) and low methoxyl pectin (LMP). The zeta potential, rheology, particle size and serum separation of AMDs were evaluated after preparing with PPP, HMP and LMP, respectively. Results indicated that PPP led to lower serum separation than LMP (14.65% for AMDs prepared with 0.5% PPP compared to 25.05% for AMDs prepared with 0.5% LMP), but still higher than HMP (9.09% for AMDs prepared with 0.5% HMP). However, narrower particle size distribution and lower viscosity of AMDs was achieved by PPP than by LMP and HMP. PPP can electrostatically adsorb on the surface of casein and its abundant neutral sugar side chains would provide steric hindrance to prevent casein flocculation in AMDs. Our results might provide some new ideas for the application of PPP in improving the stability of AMDs.


Asunto(s)
Manipulación de Alimentos/métodos , Proteínas de la Leche/química , Leche/química , Pectinas/química , Polisacáridos/química , Estabilidad Proteica , Solanum tuberosum/química , Animales , Concentración de Iones de Hidrógeno , Reología , Viscosidad
8.
Molecules ; 25(5)2020 Mar 03.
Artículo en Inglés | MEDLINE | ID: mdl-32138212

RESUMEN

Dietary intake of potato starch could induce a dramatic increase in blood glucose and is positively associated with chronic metabolic diseases (type II diabetes, cardiovascular disease, etc.). Grape seed proanthocyanidins (GSP) are known to decrease starch digestion by inhibiting digestive enzymes or changing the physicochemical properties of starch. In the present study, GSP were complexed with potato starch to prepare polyphenol-starch complexes. The physiochemical properties and digestibility of complexes were investigated by in vitro digestion model, X-ray diffraction, differential scanning calorimetry, rapid visco analyzer, Fourier transform infrared spectroscopy as well as texture profile analysis. Results indicated that the peak viscosity, breakdown, trough, and setback of the complexes disappeared, replaced by a special continuous increase in paste viscosity. The complexes showed a lower final viscosity and higher thermal stability with the increasing binding amount of GSP. GSP decreased the hardness of the complexes' gel significantly. FT-IR indicated that GSP might interact with potato starch through noncovalent forces. Additionally, the complexes also showed a higher content of slowly digestible starch and resistant starch than that of the native starch. Thus, we inferred that the addition of GSP could modify the digestibility of potato starch and be an optional way to modify the starch with lower digestion.


Asunto(s)
Extracto de Semillas de Uva/química , Proantocianidinas/química , Solanum tuberosum/química , Almidón/química , Rastreo Diferencial de Calorimetría , Espectroscopía Infrarroja por Transformada de Fourier , Difracción de Rayos X
9.
Appl Microbiol Biotechnol ; 103(17): 7055-7070, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31273395

RESUMEN

Thermostability plays an important role in the application of L-asparaginase in the pharmaceutical and food industries. Understanding the key residues and structures that influence thermostability in L-asparaginase is necessary to obtain suitable L-asparaginase candidates. In this study, special residues and structures that altered thermostability in thermophilic L-asparaginase and non-thermophilic L-asparaginase II were identified. Interchanging these special residues and structures of L-asparaginases from the four strains, that is, Pyrococcus yayanosii CH1 (PYA), Thermococcus gammatolerans (TGA), Bacillus subtilis (BSA II), and Escherichia coli (ECA II), revealed the 51st and 298th residues of PYA (corresponding to 57th, 305th residues of ECA II) as the key residues responsible for thermal stability of thermophilic L-asparaginase and non-thermophilic L-asparaginase II. Moreover, the C terminal tightness, loop rigidity, and low surface charge around activity sites were of great significance to the thermostability of L-asparaginase. This study therefore revealed the crucial amino acid residues and structures responsible for the difference in thermostability of the thermophilic and non-thermophilic L-asparaginase and provides a reference for engineering thermostability in L-asparaginase II.


Asunto(s)
Asparaginasa/química , Asparaginasa/fisiología , Secuencia de Aminoácidos , Proteínas Arqueales/química , Proteínas Arqueales/fisiología , Proteínas Bacterianas/química , Proteínas Bacterianas/fisiología , Dominio Catalítico , Biología Computacional , Estabilidad de Enzimas , Calor , Modelos Moleculares , Mutación , Conformación Proteica , Relación Estructura-Actividad
10.
Molecules ; 24(19)2019 Sep 28.
Artículo en Inglés | MEDLINE | ID: mdl-31569407

RESUMEN

Microorganisms can be used for enhancing flavors or metabolizing functional compounds. The fermented-food-derived bacterial strains comprising Bacillus velezensis, Bacillus licheniformis, and Lactobacillus reuteri mixed with Lactobacillus rhamnosus and Lactobacillus plantarum were used to ferment goji berry (Lycium barbarum L.) juice in this study. The fermentation abilities and antioxidant capacities of different mixtures of multiple strains in goji juice were compared. The results showed that the lactic acid contents increased 9.24-16.69 times from 25.30 ± 0.71 mg/100 mL in goji juice fermented using the SLV (Lactobacillus rhamnosus, Lactobacillus reuteri, and Bacillus velezensis), SZP (Lactobacillus rhamnosus, Lactobacillus plantarum, and Bacillus licheniformis), and SZVP (Lactobacillus rhamnosus, Lactobacillus plantarum, Bacillus velezensis, and Bacillus licheniformis) mixtures, and the protein contents increased 1.31-2.11 times from 39.23 ± 0.67 mg/100 mL. In addition, their contents of volatile compounds increased with positive effects on aroma in the fermented juices. Conversion of the free and bound forms of phenolic acids and flavonoids in juice was influenced by fermentation, and the antioxidant capacity improved significantly. Fermentation enhanced the contents of lactic acid, proteins, volatile compounds, and phenols. The antioxidant capacity was strongly correlated with the phenolic composition.


Asunto(s)
Antioxidantes/química , Antioxidantes/metabolismo , Bacterias/metabolismo , Fermentación , Jugos de Frutas y Vegetales , Lycium/química , Fenoles/química , Relación Estructura-Actividad , Compuestos Orgánicos Volátiles/metabolismo
11.
Mol Nutr Food Res ; 68(8): e2300643, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38600887

RESUMEN

SCOPE: Polyphenols are the major active substances in red jujube fruit, and their anti-inflammatory and antioxidant activities suggest their potential utility in the prevention of ulcerative colitis (UC). METHODS AND RESULTS: In this study, the effect of polyphenol extracts from red jujube (Ziziphus jujuba Mill. "Junzao") (PERJ) on the dextron sulfate sodium (DSS)-induced UC mice is investigated. The result shows that PERJ effectively improves clinical symptoms, including food and water intake, the disease activity insex (DAI) and spleen index, and routine blood levels, and alleviates the shortening of the colon, in mice with DSS-induced UC. Meanwhile, PERJ remarkably decreases the expression of proinflammatory factors. Moreover, PERJ repairs intestinal barrier damage by increasing the expression level of mucin 2 and mucin 3, and the result is also confirmed in the histological assessment. Besides, the expression levels of Nod-like receptor family pyrin domain-containing 3 (NLRP3) and mitogen-activated protein kinase cascade (MAPKs) signaling pathway-related proteins are inhibited by the PERJ administration. Finally, 16S rRNA sequencing analyses reveal that PERJ reverses intestinal microbiota dysbiosis by enhancing the abundance of Firmicutes and decreasing that of Proteobacteria and Bacteroidetes. CONCLUSION: PERJ probably inhibits the development of UC by suppressing the NLRP3 and MAPKs signaling pathways and regulating gut microbiota homeostasis, and can be considered as a potential resource for preventing UC.


Asunto(s)
Colitis Ulcerosa , Microbioma Gastrointestinal , Sistema de Señalización de MAP Quinasas , Extractos Vegetales , Ziziphus , Animales , Masculino , Ratones , Colitis Ulcerosa/tratamiento farmacológico , Colitis Ulcerosa/inducido químicamente , Colon/efectos de los fármacos , Colon/metabolismo , Colon/patología , Sulfato de Dextran , Microbioma Gastrointestinal/efectos de los fármacos , Homeostasis/efectos de los fármacos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Ratones Endogámicos C57BL , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Extractos Vegetales/farmacología , Polifenoles/farmacología , Ziziphus/química
12.
Mol Nutr Food Res ; 68(4): e2300334, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38150643

RESUMEN

SCOPE: Chronic liver diseases are clinically silent and responsible for significant morbidity and mortality worldwide. Jujube has displayed various biological activities. Here, the therapeutic effect of Lactobacillus acidophilus (L. acidophilus)-fermented jujube juice (FJJ) and the possible mechanism against chronic liver injury (CLI) in mice are further studied. METHODS AND RESULTS: After the CCl4 -induced CLI mice are separately treated with L. acidophilus (LA), unfermented jujube juice (UFJJ), and FJJ, FJJ but not LA or UFJJ suppresses the liver index. By using H&E staining, immunofluorescence staining, RT-PCR, and western blotting, it is shown that LA, UFJJ, and FJJ intervention ameliorate hepatocyte necrosis, inhibit the mRNA levels of pro-inflammatory (NLRP3, Caspase-1, IL-1ß, and TNF-α) and fibrosis-associated factors (TGF-ß1, LXRα, and MMP2). Also, FJJ displays significant protection against mucosal barrier damage in CLI mice. Among the three interventions, FJJ exhibits the best therapeutic effect, followed by UFJJ and LA. Furthermore, FJJ improves dysbiosis in CLI mice. CONCLUSIONS: This study suggests that FJJ exhibits a protective effect against CCl4 -induced CLI mice by inhibiting apoptosis and oxidative stress, regulating liver lipid metabolism, and improving gut microecology. Jujube juice fermentation with L. acidophilus can be a food-grade supplement in treating CLI and related liver diseases.


Asunto(s)
Hepatopatías , Ziziphus , Ratones , Animales , Lactobacillus acidophilus/metabolismo , Apoptosis
13.
ACS Omega ; 9(9): 10276-10285, 2024 Mar 05.
Artículo en Inglés | MEDLINE | ID: mdl-38463329

RESUMEN

l-threonine as an important precursor substance of l-isoleucine and improving its accumulation in Escherichia coli became an important idea to construct a chassis strain with high l-isoleucine production. Meanwhile, the effect of l-threonine metabolic pathway disruption in E. coli for the improved production of l-isoleucine remains unrevealed. In the present study, a mutant strain of E. coli was engineered by inactivating specific metabolic pathways (e.g., Δtdh, ΔltaE, and ΔyiaY) that were associated with l-threonine metabolism but unrelated to l-isoleucine synthesis. This was done with the aim to reduce the breakdown of l-threonine and, thereby, increase the production of l-isoleucine. The results obtained demonstrated a 72.3% increment in l-isoleucine production from 4.34 to 7.48 g·L-1 in the mutant strain compared with the original strain, with an unexpected 10.3% increment in bacterial growth as measured at OD600. Transcriptome analysis was also conducted on both the mutant strain NXU102 and the original strain NXU101 in the present study to gain a comprehensive understanding of their physiological attributes. The findings revealed a notable disparity in 1294 genes between the two strains, with 658 genes exhibiting up-regulation and 636 genes displaying down-regulation. The activity of tricarboxylic acid (TCA) cycle-related genes was found to decrease, but oxidative phosphorylation-related genes were highly up-regulated, which explained the increased activity of the mutant strain. For instance, l-lysine catabolism-related genes were found to be up-regulated, which reconfigured the carbon flow into the TCA cycle. The augmentation of acetic acid degradation pathway-related genes assisted in the reduction in acetic acid accumulation that could retard cell growth. Notably, substantial up-regulation of the majority of genes within the aspartate pathway could potentially account for the increased production of l-isoleucine in the present study. In this paper, a chassis strain with an l-isoleucine yield of 7.48 g·L-1 was successfully constructed by cutting off the threonine metabolic pathway. Meanwhile, transcriptomic analysis revealed that the cutting off of the threonine metabolic pathway induced perturbation of genes related to the pathways associated with the synthesis of l-isoleucine, such as the tricarboxylic acid cycle, glycolysis, and aspartic acid pathway.

14.
Can J Microbiol ; 59(6): 374-9, 2013 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-23750951

RESUMEN

A major problem when pyrimidine de novo biosynthesis is used for cytidine production is the existence of many negative regulatory factors. Cytidine biosynthesis in Bacillus amyloliquefaciens proceeds via a pathway that is controlled by uridine monophosphate (UMP) through feedback inhibition of carbamoyl phosphate synthetase (CPS), the enzyme that converts CO2, NH3, and glutamine to carbamoyl phosphate. In this study, the gene carB encoding the large subunit of CPS from B. amyloliquefaciens CYT1 was site directed, and the UMP binding sites of feedback inhibition in Bam-CPS are described. The residues Thr-941, Thr-970, and Lys-986 in CPS from B. amyloliquefaciens were subjected to site-directed mutagenesis to alter UMP's feedback inhibition of CPS. To find feedback-resistant B. amyloliquefaciens, the influence of the T941F, T970A, K986I, T941F/K986I, and T941F/T970A/K986I mutations on CPS enzymatic properties was studied. The recombinant B. amyloliquefaciens with mutated T941F/K986I and T941F/T970A/K986I CPS showed a 3.7- and 5.7-fold increase, respectively, in cytidine production in comparison with the control expressing wild-type CPS, which was more suitable for further application of the cytidine synthesis. To a certain extent, the 5 mutations were found to release the enzyme from UMP inhibition and to improve B. amyloliquefaciens cytidine-producing strains.


Asunto(s)
Bacillus/enzimología , Bacillus/genética , Carbamoil-Fosfato Sintasa (Glutamina-Hidrolizante)/genética , Carbamoil-Fosfato Sintasa (Glutamina-Hidrolizante)/metabolismo , Citidina/biosíntesis , Uridina Monofosfato/metabolismo , Secuencia de Aminoácidos , Bacillus/metabolismo , Sitios de Unión/genética , Carbamoil-Fosfato Sintasa (Glutamina-Hidrolizante)/química , Carbamoil-Fosfato Sintasa (Glutamina-Hidrolizante)/aislamiento & purificación , Carbamoil Fosfato/metabolismo , Retroalimentación Fisiológica , Glutamina/metabolismo , Cinética , Modelos Moleculares , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Conformación Proteica , Subunidades de Proteína/química , Subunidades de Proteína/genética , Subunidades de Proteína/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo
15.
Biotechnol Lett ; 35(2): 245-51, 2013 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-23070626

RESUMEN

Cytidine is a precursor of several antiviral drugs. The pentose phosphate pathway (PPP) is primarily responsible for NADPH and 5-phospho-α-D-ribose 1-diphosphate as an important precursor of cytidine biosynthesis in Escherichia coli. To enhance cytidine production, we obtained the recombinant E. coli CYT15-gnd-prs-zwf that co-expressed the prs, zwf, and gnd genes encoding phosphoribosylpyrophosphate synthetase, glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase (three key enzymes in PPP) respectively. In fermentation experiments, strain CYT15-gnd-prs-zwf produced 735 mg cytidine/l using glucose as substrate, which was approx. 128 % higher than the cytidine production by the parental strain (CYT15). Co-expression of zwf, gnd, and prs decreased growth (3.2 %) slightly and increased glucose uptake (72 %). This is the first study to report increased cytidine production by increasing metabolic flux through the PPP in E. coli.


Asunto(s)
Citidina/biosíntesis , Escherichia coli/genética , Escherichia coli/metabolismo , Ingeniería Metabólica , Redes y Vías Metabólicas/genética , Fermentación , Expresión Génica , Glucosafosfato Deshidrogenasa/genética , Glucosafosfato Deshidrogenasa/metabolismo , Fosfogluconato Deshidrogenasa/genética , Fosfogluconato Deshidrogenasa/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Ribosa-Fosfato Pirofosfoquinasa/genética , Ribosa-Fosfato Pirofosfoquinasa/metabolismo
16.
Biosensors (Basel) ; 13(9)2023 Sep 17.
Artículo en Inglés | MEDLINE | ID: mdl-37754120

RESUMEN

Electrochemical biosensors based on immobilized enzymes are among the most popular and commercially successful biosensors. The literature in this field suggests that modification of electrodes with nanomaterials is an excellent method for enzyme immobilization, which can greatly improve the stability and sensitivity of the sensor. However, the poor stability, weak reproducibility, and limited lifetime of the enzyme itself still limit the requirements for the development of enzyme electrochemical biosensors for food production process monitoring. Therefore, constructing sensing technologies based on enzyme electrochemical biosensors remains a great challenge. This article outlines the construction principles of four generations of enzyme electrochemical biosensors and discusses the applications of single-enzyme systems, multi-enzyme systems, and nano-enzyme systems developed based on these principles. The article further describes methods to improve enzyme immobilization by combining different types of nanomaterials such as metals and their oxides, graphene-related materials, metal-organic frameworks, carbon nanotubes, and conducting polymers. In addition, the article highlights the challenges and future trends of enzyme electrochemical biosensors, providing theoretical support and future perspectives for further research and development of high-performance enzyme chemical biosensors.

17.
Int J Biol Macromol ; 235: 123829, 2023 Apr 30.
Artículo en Inglés | MEDLINE | ID: mdl-36858086

RESUMEN

Heat-moisture treatment (HMT) and annealing (ANN) were applied to modify the proso millet starch, and then the physicochemical properties as well as the in vitro digestion of the modified starch were investigated systematically. Results indicated that HMT and ANN did not change the typical A-type crystallinity. However, both processes cause cracks and dents on the surface of the granule. The gelatinization temperature increased while peak viscosity value, relative crystallinity and gelatinization enthalpy of proso millet starch decreased significantly after HTM and ANN. Meanwhile, a remarkable increase of the slowly digestible starch(SDS) and resistant starch(RS) content was noticed after HTM and ANN modification (the highest content of SDS and RS after HTM and ANN were 9.52 ± 0.82 %, 12.03 ± 1.36 % and 12.15 ± 0.89 %, 8.75 ± 1.63 %, respectively). Those results indicated that the ANN and HMT processes could modify the physicochemical properties and in vitro digestion of proso millet starch efficiently and showed potential application to produce healthy starch food with lower digestion.


Asunto(s)
Panicum , Almidón , Almidón/química , Calor , Temperatura , Harina/análisis
18.
ACS Appl Mater Interfaces ; 15(15): 18569-18589, 2023 Apr 19.
Artículo en Inglés | MEDLINE | ID: mdl-37037009

RESUMEN

The decreased number of viable bacteria and the ability of Bifidobacterium to adhere to and colonize the gut in the gastrointestinal environment greatly limit their efficacy. To solve this problem, thiolated carboxymethyl cellulose sodium (CMC) probiotic double-layered multinucleated microcapsules with Bifidobacterium adolescentis FS2-3 in the inner layer and Bacillus subtilis SN15-2 embedded in the outer layers were designed. First, the viable counts and release rates of microcapsules were examined by in vitro simulated digestion assays, and it was found that microcapsules were better protected from gastrointestinal digestion than the controls. Compared with free Bifidobacterium strains, double-layered multinucleated microcapsules have higher viable bacterial survival rates and storage stability. Second, through in vitro rheology, tensile tests, isotherm titration calorimetry, and adhesion tests, it was observed that thiolated CMC could enhance the strong interaction of Bifidobacterium with intestinal mucus and significantly promote the proliferation and growth of probiotics. Finally, double-layered multinucleated microcapsules containing B. adolescentis FS2-3 and B. subtilis SN15-2 modified with sulfhydryl-modified CMC were studied in the intestine. Alleviation of Escherichia coli O157:H7 induced intestinal inflammation. The results showed that microencapsulation could significantly increase the colon content of Bifidobacterium, relieve intestinal inflammation symptoms in mice with bacterial enteritis, and repair the intestinal microbiota disorder caused by inflammation. The probiotic double-layered multinucleated microcapsules prepared in this study can improve the survival rate of probiotics and promote proliferation, adhesion, and colonization of probiotics.


Asunto(s)
Escherichia coli O157 , Probióticos , Animales , Ratones , Carboximetilcelulosa de Sodio , Cápsulas/química , Bifidobacterium , Probióticos/uso terapéutico , Sodio
19.
Food Chem X ; 14: 100339, 2022 Jun 30.
Artículo en Inglés | MEDLINE | ID: mdl-35634223

RESUMEN

In order to investigate the effect of different cooking methods on the phytochemicals and antioxidant activities of potato from different varieties, three varieties of potatoes were cooked with seven domestic methods. The contents of total phenolic, total carotenoid, vitamin C and phenolic acids of cooked potato were analyzed as well as the antioxidant activities. Results indicated that all the cooking methods showed negative effects on the contents of vitamin C, total phenolic, phenolic acids and DPPH radical-scavenging activity, but the effects were depended on the cooking methods, as frying, air-drying and roasting showed a more intensive decrease of total phenolic, phenolic acids and antioxidant activities than that of steaming or microwaving, regardless of the potato verities. From the perspective of remaining phytochemicals and antioxidant activities, Zhongshubahao might be an ideal potato sample and steaming or microwaving were optional methods for cooking potatoes.

20.
Nat Commun ; 13(1): 6690, 2022 11 05.
Artículo en Inglés | MEDLINE | ID: mdl-36335132

RESUMEN

The Allium genus is cultivated globally as vegetables, condiments, or medicinal plants and is characterized by large genomes and strong pungency. However, the genome evolution and genomic basis underlying their unique flavor formation remain poorly understood. Herein, we report an 11.27-Gb chromosome-scale genome assembly for bunching onion (A. fistulosum). The uneven bursts of long-terminal repeats contribute to diversity in genome constituents, and dispersed duplication events largely account for gene expansion in Allium genomes. The extensive duplication and differentiation of alliinase and lachrymatory factor synthase manifest as important evolutionary events during flavor formation in Allium crops. Furthermore, differential selective preference for flavor-related genes likely lead to the variations in isoalliin content in bunching onions. Moreover, we reveal that China is the origin and domestication center for bunching onions. Our findings provide insights into Allium genome evolution, flavor formation and domestication history and enable future genome-assisted breeding of important traits in these crops.


Asunto(s)
Allium , Allium/genética , Cebollas/genética , Cromosomas de las Plantas/genética , Fitomejoramiento , Evolución Molecular
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