Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 100
Filtrar
1.
Nat Chem Biol ; 18(8): 886-893, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35817967

RESUMO

Although natural products and synthetic small molecules both serve important medicinal functions, their structures and chemical properties are relatively distinct. To expand the molecular diversity available for drug discovery, one strategy is to blend the effective attributes of synthetic and natural molecules. A key feature found in synthetic compounds that is rare in nature is the use of fluorine to tune drug behavior. We now report a method to site-selectively incorporate fluorine into complex structures to produce regioselectively fluorinated full-length polyketides. We engineered a fluorine-selective trans-acyltransferase to produce site-selectively fluorinated erythromycin precursors in vitro. We further demonstrated that these analogs could be produced in vivo in Escherichia coli on engineering of the fluorinated extender unit pool. By using engineered microbes, elaborate fluorinated compounds can be produced by fermentation, offering the potential for expanding the identification and development of bioactive fluorinated small molecules.


Assuntos
Produtos Biológicos , Policetídeos , Aciltransferases/metabolismo , Produtos Biológicos/química , Escherichia coli/genética , Escherichia coli/metabolismo , Flúor , Policetídeos/química
2.
Inorg Chem ; 63(6): 3199-3206, 2024 Feb 12.
Artigo em Inglês | MEDLINE | ID: mdl-38286822

RESUMO

A deep insight into surface structural evolution of the catalyst is a challenging issue to reveal the structure-activity relationship. In this contribution, based on a surface alloying strategy, the dual-functional Pd@NiPd catalyst with a unique core-shell hierarchical structure is developed through selective crystal growth, surface cocrystallization, directional self-assembly, and reduction process. The surface defects are created in situ on the outer NiPd alloy layer in the electrochemical redox processes, which endow the Pd@NiPd catalyst with excellent electrocatalytic activity of hydrogen generation reaction (HER) and oxygen generation reaction (OER) in alkaline media. The optimal Pd@NiPd-2 catalyst requires an overpotential of only 18 mV that is far lower than Pt/C benchmark (43 mV) at the current density of 10 mA cm-2 for the HER, and 210 mV that is far lower than RuO2 benchmark (430 mV) at 50 mA cm-2 for the OER. Density functional theory (DFT) calculations reveal that the outstanding electrocatalytic activity is originated from the creation of surface defect structure that induces a significant reduction in the adsorption and dissociation energy barriers of H2O molecules in the HER and a decrease in the conversion energy from O* to OOH* that resulted from the synergy of two adjacent Pd sites by forming O-bridge. This work affords a typical paradigm for exploiting efficient catalysts and investigating the dependence of electrocatalytic activity on the surface structural evolution.

3.
Lipids Health Dis ; 23(1): 205, 2024 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-38951804

RESUMO

BACKGROUND: Glycerophospholipids (GPLs) are essential for cell membrane structure and function. Sphingomyelin and its metabolites regulate cell growth, apoptosis, and stress responses. This study aimed to investigate lipid metabolism in patients experiencing sudden sensorineural hearing loss across all frequencies (AF-SSNHL). METHODS: The study included 60 patients diagnosed with unilateral AF-SSNHL, among whom 30 patients had a level of hearing improvement ≥ 15 dB after 6 months of follow-up. A propensity score-matched (2:1) control group was used. Liquid chromatography‒mass spectrometry based untargeted lipidomics analysis combined with multivariate statistics was performed to investigate the lipids change. The "lipidome" R package and weighted gene co-expression network analysis (WGCNA) were utilised to assess the lipids' structural features and the association between lipids and hearing. RESULTS: Lipidomics successfully differentiated the AF-SSNHL group from the control group, identifying 17 risk factors, mainly including phosphatidylcholine (PC), phosphatidylethanolamine (PE), and related metabolites. The ratios of lysophosphatidylcholine/PC, lysophosphatidylethanolamine/PE, and lysodimethylphosphatidylethanolamine/PE were upregulated, while some glycerophospholipid (GPL)-plasmalogens were downregulated in the AF-SSNHL group, indicating abnormal metabolism of GPLs. Trihexosylceramide (d34:1), PE (18:1e_22:5), and sphingomyelin (d40:3) were significantly different between responders and nonresponders, and positively correlated with hearing improvement. Additionally, the results of the WGCNA also suggested that partial GPL-plasmalogens were positively associated with hearing improvement. CONCLUSION: AF-SSNHL patients exhibited abnormally high blood lipids and pronounced GPLs metabolic abnormalities. Sphingolipids and GPL-plasmalogens had an association with the level of hearing improvement. By understanding the lipid changes, clinicians may be able to predict the prognosis of hearing recovery and personalize treatment approaches.


Assuntos
Biomarcadores , Perda Auditiva Neurossensorial , Metabolismo dos Lipídeos , Lipidômica , Humanos , Feminino , Masculino , Pessoa de Meia-Idade , Biomarcadores/sangue , Perda Auditiva Neurossensorial/sangue , Adulto , Perda Auditiva Súbita/sangue , Glicerofosfolipídeos/sangue , Idoso , Fosfatidiletanolaminas/sangue , Fosfatidiletanolaminas/metabolismo , Fosfatidilcolinas/sangue , Fosfatidilcolinas/metabolismo , Lisofosfatidilcolinas/sangue , Esfingomielinas/sangue , Esfingomielinas/metabolismo , Lisofosfolipídeos
4.
Inorg Chem ; 62(41): 16954-16964, 2023 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-37787454

RESUMO

It has been a challenging issue to profoundly actuate the transfer and separation of photoinduced charge carriers by controlling the interface structure inside the heterojunction, owing to the molecular/subnanometric level interface region. Herein, a unique one-dimensional/two-dimensional (1D/2D) CoTe/PCN Z-scheme heterojunction is fabricated through the self-assembly of CoTe nanorods on the surface of polymeric carbon nitride (PCN) nanosheets. Significantly, in situ N-doping in the molecular/subnanometric surface oxidized layer of CoTe nanorods is achieved, effectively adjusting its chemical structure and element chemical states. Moreover, this N-doped surface oxidized layer can serve as a recombination region of photogenerated electrons from PCN and photogenerated holes from CoTe to increase the overall carrier separation efficiency in the Z-scheme heterojunction actuated by the built-in electric field. As a result, the photocatalytic CO2 reduction (CO2R) performance is enhanced dramatically, in which the yield of CO generated over the optimal 1D/2D CoTe/PCN heterojunction reaches up to triple than that over PCN. This unique contribution provides an emblematic paradigm for adjusting the interfacial structure of heterojunction and has a profound insight into the interfacial adjusting mechanism to improve the charge separation efficiency in the photocatalytic reaction.

5.
BMC Public Health ; 23(1): 1633, 2023 08 25.
Artigo em Inglês | MEDLINE | ID: mdl-37626323

RESUMO

BACKGROUND: The knowledge, attitude, and practice (KAP) of Chinese patients with allergic rhinitis (AR) on AR is poorly known. This study investigated the KAP towards AR in patients with this disease and explored the factors associated with KAP. METHODS: This cross-sectional study enrolled patients with AR in Zhangjiagang Hospital of Traditional Chinese Medicine between October 2022 and March 2023. RESULTS: This study included 656 valid questionnaires. Most participants were 26-35 years old (36.13%) and were female (55.18%). The knowledge, attitude, and practice scores were 5.70 ± 2.88 (possible range: 0-12), 29.51 ± 3.52 (possible range: 9-45), and 34.13 ± 7.55 (possible range: 9-45), indicating poor knowledge, unfavorable attitudes, and proactive practice. AR history of 3-5 years (adjusted odds ratio (adjOR) = 1.62, 95% confidence interval (CI): 1.03-2.54, P = 0.037), AR history of > 6 years (adjOR = 1.64, 95%CI: 1.06-2.54, P = 0.027), and know their own allergens (adjOR = 2.34, 95%CI: 1.28-4.25, P = 0.005) were independently associated with the sufficient knowledge. AR history of ≥ 6 years (adjOR = 0.60, 95%CI: 0.37-0.96, P = 0.035), and liking sports (adjOR = 1.58, 95%CI = 1.07-2.33, P = 0.020) were independently associated with the positive attitude. The knowledge scores (adjOR = 1.14, 95%CI: 1.05-1.22, P = 0.001), attitude scores (adjOR = 1.24, 95%CI: 1.17-1.32, P < 0.001), age 36-45 (adjOR = 2.13, 95%CI: 1.19-3.82, P = 0.011), employed (adjOR = 0.59, 95%CI: 0.37-0.94, P = 0.026), and liking sports (adjOR = 2.11, 95%CI: 1.43-3.14, P < 0.001) were independently associated with the proactive practice. CONCLUSIONS: Patients with AR have poor knowledge and unfavorable attitudes but good practice toward AR. Continuous quality teaching interventions and education on patients for AR were recommended.


Assuntos
Conhecimentos, Atitudes e Prática em Saúde , Rinite Alérgica , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Povo Asiático , Estudos Transversais , Rinite Alérgica/epidemiologia , Rinite Alérgica/terapia
6.
Inorg Chem ; 61(34): 13453-13461, 2022 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-35969492

RESUMO

It is a challenge to fabricate atomically dispersed metal clusters in polymeric carbon nitride (PCN) for durable photocatalytic reactions owing to the thermodynamic stability limitation. Herein, atomically dispersed Ru clusters are implanted into the PCN skeleton matrix based on an ionic diffusion and coordination (IDC) strategy, the stability of which is improved owing to the robust Ru-N bonds in the formed RuN4 and RuN3 configurations. Additionally, RuN4 and RuN3 as charge transport bridges between two adjacent melon strands efficaciously conquer hydrogen bond restriction in the skeleton to facilitate the in-plane mobility and separation of charge carriers. Moreover, the synergistic effect of adjacent Ru atoms is triggered on the assembled RuN3-RuN4 and RuN3-RuN3 in the atomically dispersed Ru clusters to significantly decrease hydrogen adsorption energy. As a result, the optimal PCN-Ru photocatalyst achieves nearly 6 times higher than the photocatalytic hydrogen evolution (PHE) rate of the Pt/PCN benchmark and maintains the long-term stable running for 104 h of 26 cycles; its overall PHE performance is far superior to the most of single atoms supported on g-C3N4 photocatalysts reported. The findings here gain new insight into the preparation strategy, structure configuration, and reaction mechanism for atomically dispersed metal clusters supported on PCN, which further stimulates the intensive investigations toward developing more efficient and stable PCN-like photocatalytic materials.

7.
Inorg Chem ; 61(39): 15600-15606, 2022 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-36134910

RESUMO

Micro/nanostructure control of heterostructures is still a challenge for achieving high efficiency and selectivity of photocatalytic CO2 conversion. In this work, a new three-dimensiona/two-dimensional (3D/2D) heterostructure is fabricated by encapsulating RuS2 nanospheres in the interlayer of mesoporous polymeric carbon nitride (PCN) nanosheets based on an in situ growth and polymerization strategy. The unique microstructure of the obtained 3D/2D RuS2/PCN heterojunction can effectively improve the transfer and separation efficiency of photogenerated charge carriers, reduce the mass transfer resistance of CO2 toward active sites, and provide a confined reaction space, thus propelling the photocatalytic CO2 reduction to CO with high selectivity. The CO yield over the optimal 5%-RuS2/PCN sample reaches 4.2 and 2.8 times as high as that of single PCN and RuS2 within 4 h, respectively. Furthermore, the plausible charge transfer mechanism and CO2 reduction path are revealed by time-dependent in situ Fourier transform infrared (FT-IR) spectra combined with photophysical, electrochemical, and photoelectrochemical techniques and density functional theory (DFT) calculations. This work develops the microstructural engineering design strategy of PCN-based heterojunctions for selective photocatalytic CO2 fuel conversion.

8.
Microb Cell Fact ; 20(1): 149, 2021 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-34325704

RESUMO

BACKGROUND: The intracellular ATP level is an indicator of cellular energy state and plays a critical role in regulating cellular metabolism. Depletion of intracellular ATP in (facultative) aerobes can enhance glycolysis, thereby promoting end product formation. In the present study, we examined this s trategy in anaerobic ABE (acetone-butanol-ethanol) fermentation using Clostridium acetobutylicum DSM 1731. RESULTS: Following overexpression of atpAGD encoding the subunits of water-soluble, ATP-hydrolyzing F1-ATPase, the intracellular ATP level of 1731(pITF1) was significantly reduced compared to control 1731(pIMP1) over the entire batch fermentation. The glucose uptake was markedly enhanced, achieving a 78.8% increase of volumetric glucose utilization rate during the first 18 h. In addition, an early onset of acid re-assimilation and solventogenesis in concomitant with the decreased intracellular ATP level was evident. Consequently, the total solvent production was significantly improved with remarkable increases in yield (14.5%), titer (9.9%) and productivity (5.3%). Further genome-scale metabolic modeling revealed that many metabolic fluxes in 1731(pITF1) were significantly elevated compared to 1731(pIMP1) in acidogenic phase, including those from glycolysis, tricarboxylic cycle, and pyruvate metabolism; this indicates significant metabolic changes in response to intracellular ATP depletion. CONCLUSIONS: In C. acetobutylicum DSM 1731, depletion of intracellular ATP significantly increased glycolytic rate, enhanced solvent production, and resulted in a wide range of metabolic changes. Our findings provide a novel strategy for engineering solvent-producing C. acetobutylicum, and many other anaerobic microbial cell factories.


Assuntos
Trifosfato de Adenosina/metabolismo , Clostridium acetobutylicum/metabolismo , Fermentação , Glicólise , Solventes/metabolismo , Acetona/metabolismo , Anaerobiose , Biocombustíveis , Butanóis/metabolismo , Clostridium acetobutylicum/genética , Etanol/metabolismo , Hidrólise
9.
AIDS Res Ther ; 18(1): 14, 2021 04 20.
Artigo em Inglês | MEDLINE | ID: mdl-33879191

RESUMO

BACKGROUND: HIV testing and early linkage to care are critical for reducing the risk of HIV transmission. HIV self-testing (HIVST) is a useful tool for increasing HIV testing frequency.This study aimed to investigate HIVST rates among men who have sex with men (MSM), the characteristics of MSM who had HIVST, and factors associated with HIVST uptake among MSM in Ningbo, China. METHODS: A cross-sectional study was conducted from April to October 2019 in Ningbo,China. Participants were aged at least 18 years and having had sexual contact with men in the past year. Proportions were used for categorical variables. Adjusted Odds Ratio (AOR) and 95% Confidence Interval (CI) for characteristics associated with HIVST uptake was processed by multivariable logistic regression models. RESULTS: Among a sample of 699 MSM recruited, 38.2% had reported previous use of an HIV self-test kit. A greater proportion of HIVST users had a higher frequency of HIV testing (≥ 2 times: 70.0% versus 41.2%, p < 0.001) in the past 1 year. The odds of older age (30-39 years: AOR = 0.49, CI 0.32-0.76; more than 40 years: AOR = 0.07, CI 0.04-0.14, compared to 18-29 years), bisexual (AOR = 0.49, CI 0.29-0.84) were lower among HIVST users,and were higher among MSM who were higher education level (high school: AOR = 2.82, CI 1.70-4.69, compared to middle school or less), gay apps use (AOR = 1.86, CI 1.13-3.05), multiple male sex partners (AOR = 1.90, CI 1.29-2.80), frequency of male-male sexual contact ≥ 1 times per week (AOR = 1.86, CI 1.30-2.66), syphilis infection (AOR = 5.48, CI 2.53-11.88). CONCLUSIONS: Further HIVST education should be strengthened for school-aged children and teenagers, and free HIVST kits may be provided to high-risk MSM through gay apps and CBO to achieve the increased HIV testing frequency.


Assuntos
Infecções por HIV , Minorias Sexuais e de Gênero , Adolescente , Idoso , Criança , China/epidemiologia , Estudos Transversais , Infecções por HIV/diagnóstico , Infecções por HIV/epidemiologia , Homossexualidade Masculina , Humanos , Masculino , Prevalência , Autoteste
10.
BMC Public Health ; 21(1): 1118, 2021 06 10.
Artigo em Inglês | MEDLINE | ID: mdl-34112128

RESUMO

BACKGROUND: In China, enterovirus 71 (EV71) is the major etiological agents of hand foot mouth disease that poses severe risks to children's health. Since 2015, three inactivated EV71 vaccines have been approved for use. Previous studies indicated the high willingness of EV71 vaccination in eastern China. However, few studies have assessed coverage and utilization patterns of EV71 vaccine in China. METHODS: Children born during 2012-2018 were sampled and their records were abstracted from Ningbo childhood immunization information management system. Descriptive statistics characterized the study population and assessed coverage and timeliness for EV71 vaccination. Simultaneous administration patterns as well as type of EV71 vaccine used were also evaluated. Bivariate and multivariable analysis was used to examine the relationship of socio-demographic characteristics with vaccination coverage and timeliness. RESULTS: Of 716,178 children living in Ningbo. One hundred seventy-two thousand two hundred thirty-six received EV71 vaccine with a coverage rate of 24.05% and only 8.61% received vaccination timely. 21.97% of children received the complete two dose EV71 series but only 6.49% completed timely. Vaccination coverage and timeliness increased significantly from 2012 birth cohort to 2018 birth cohort. Relatively higher coverage and timeliness were observed in resident children, Inner districts, high socioeconomic areas and large-scaled immunization clinics. Of 329,569 doses of EV71 vaccine, only 5853(1.78%) doses were administered at the same day as other vaccines. CONCLUSIONS: There is a need for increasing EV71 vaccination coverage and timeliness as well as eliminating disparities among different populations. Our study highlights the importance of simultaneous administration to increasing coverage and timeliness of EV71 vaccination.


Assuntos
Enterovirus Humano A , Enterovirus , Vacinas Virais , Criança , China , Humanos , Lactente , Vacinação , Cobertura Vacinal
11.
Am J Otolaryngol ; 42(3): 102922, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33454456

RESUMO

PURPOSE: To evaluate the effectiveness of systemic Ginkgo biloba diterpene lactone therapy for sudden sensorineural hearing loss. METHODS: This retrospective review investigated 56 patients with unilateral sudden sensorineural hearing loss. Among them, 26 patients received conventional therapy (group C, intravenous methylprednisolone), and 30 received conventional therapy supplemented with Ginkgo biloba diterpene lactone (group G). Pure tone audiometry was measured before treatment and 1 month after treatment. The average pure tone audiometry gain, pure tone audiometry gain at each frequency, pure tone audiometry gain according to initial hearing loss, and rate of effectiveness were defined as functionally relevant recovery of hearing and compared between the two groups. RESULTS: The average pure tone audiometry gain was significantly greater in group G (20.6 ± 15.1 dB) than in group C (11.9 ± 13.3 dB) (p = 0.025), with similar trends at 250, 1 k, and 8 k Hz. In the subgroup of patients with profound hearing loss (initial pure tone audiometry >70 dB), hearing gain was significantly higher in group G (26.7 ± 14.4 dB) than in C (5.5 ± 9.0 dB) (p = 0.034). In the mild-moderate hearing loss subgroup (initial pure tone audiometry ≤70 dB), the pure tone audiometry gain did not differ significantly (group G: 18.4 ± 14.3 dB; group C: 13.0 ± 13.4 dB) (p = 0.209). The overall rate of effectiveness was 73.3% and 57.7% in groups G and C, respectively; however, the difference was statistically insignificant (p = 0.218). CONCLUSIONS: Compared with conventional therapy alone, supplementary systemic administration of Ginkgo biloba diterpene lactone to treat sudden sensorineural hearing loss could improve hearing recovery, especially, in patients with profound hearing loss.


Assuntos
Diterpenos/administração & dosagem , Ginkgo biloba/química , Perda Auditiva Neurossensorial/tratamento farmacológico , Perda Auditiva Súbita/tratamento farmacológico , Lactonas/administração & dosagem , Fitoterapia , Adulto , Audiometria de Tons Puros , Diterpenos/isolamento & purificação , Quimioterapia Combinada , Feminino , Perda Auditiva Neurossensorial/diagnóstico , Perda Auditiva Súbita/diagnóstico , Humanos , Lactonas/isolamento & purificação , Masculino , Metilprednisolona/administração & dosagem , Pessoa de Meia-Idade , Resultado do Tratamento
12.
J Am Chem Soc ; 141(42): 16877-16883, 2019 10 23.
Artigo em Inglês | MEDLINE | ID: mdl-31547647

RESUMO

Polymers are an important class of materials that are used for a broad range of applications, from drug delivery to packaging. Given their widespread use, a major challenge in this area is the development of technology for their production from renewable sources and efforts to promote their efficient recycling and biodegradation. In this regard, the synthesis of polyesters based on the natural polyhydroxyalkanoate (PHA) pathway offers an attractive route for producing sustainable polymers. However, monomer diversity in naturally occurring polyesters can be limited with respect to the design of polymers with material properties suitable for various applications. In this work, we have engineered a pathway to produce α-methyl-branched PHA. In the course of this work, we have also identified a PHA polymerase (CapPhaEC) from activated sludge from wastewater treatment that demonstrates a higher capacity for incorporation of α-branched monomer units than those previously identified or engineered. Production in Escherichia coli allows the construction of microbial strains that produce the copolyesters with 21-36% branched monomers using glucose and propionate as carbon sources. These polymers have typical weight-average molar masses (Mw) in the range (1.7-2.0) × 105 g mol-1 and display no observable melting transition, only relatively low glass transition temperatures from -13 to -20 °C. The lack of a melting transition indicates that these polymers are amorphous materials with no crystallinity, which is in contrast to the natural poly(hydroxybutyrate) homopolymer. Our results expand the utility of PHA-based pathways and provide biosynthetic access to α-branched polyesters to enrich the properties of bio-based sustainable polymers.


Assuntos
Aciltransferases/genética , Aciltransferases/metabolismo , Poliésteres/química , Engenharia de Proteínas , Aciltransferases/química , Modelos Moleculares , Conformação Proteica
13.
J Am Chem Soc ; 139(41): 14526-14532, 2017 10 18.
Artigo em Inglês | MEDLINE | ID: mdl-28990776

RESUMO

Cell-based synthesis offers many opportunities for preparing small molecules from simple renewable carbon sources by telescoping multiple reactions into a single fermentation step. One challenge in this area is the development of enzymatic carbon-carbon bond forming cycles that enable a modular disconnection of a target structure into cellular building blocks. In this regard, synthetic pathways based on thiolase enzymes to catalyze the initial carbon-carbon bond forming step between acyl coenzyme A (CoA) substrates offer a versatile route for biological synthesis, but the substrate diversity of such pathways is currently limited. In this report, we describe the identification and biochemical characterization of a thiolase-ketoreductase pair involved in production of branched acids in the roundworm, Ascaris suum, that demonstrates selectivity for forming products with an α-methyl branch using a propionyl-CoA extender unit. Engineering synthetic pathways for production of α-methyl acids in Escherichia coli using these enzymes allows the construction of microbial strains that produce either chiral 2-methyl-3-hydroxy acids (1.1 ± 0.2 g L-1) or branched enoic acids (1.12 ± 0.06 g L-1) in the presence of a dehydratase at 44% and 87% yield of fed propionate, respectively. In vitro characterization along with in vivo analysis indicates that the ketoreductase is the key driver for selectivity, forming predominantly α-branched products even when paired with a thiolase that highly prefers unbranched linear products. Our results expand the utility of thiolase-based pathways and provide biosynthetic access to α-branched compounds as precursors for polymers and other chemicals.


Assuntos
Acetil-CoA C-Acetiltransferase/metabolismo , Hidroxiácidos/metabolismo , Escherichia coli/metabolismo , Hidroliases/metabolismo , Propionatos/metabolismo
14.
Metab Eng ; 44: 284-292, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-29102594

RESUMO

Biotechnological production of butanol in heterologous hosts has recently attracted many interests. Of the heterologous hosts investigated to date, engineered Escherichia coli has shown a superior butanol yield than the natural butanol-producing clostridial strains. However, all reported butanol-producing E. coli strains contain vectors and inducible promoters, which means antibiotics and inducers are required in the fermentation. The aim of this study was to develop a completely chromosomally engineered E. coli strain capable of producing butanol efficiently in the absence of vectors, antibiotics, and inducers. The challenges are the expression strength of chromosomally engineered genes under constitutive promoters is much weaker than the vector engineered genes under inducible promoters. To address these challenges, the butanol pathway was engineered into the chromosome in the first place, then the host and the butanol pathway was iteratively engineered through rational and non-rational strategies to develop an efficient butanol producer where the heterologous butanol pathway fits the host well. Finally, a systematically chromosomally engineered E. coli strain EB243, in which 33 native genes were deleted and 5 heterologous genes were introduced, was developed. Strain EB243 could produce 20g/L butanol with a yield of 34% (w/w, 83% of theoretical yield) in batch fermentation without any antibiotics and inducers, thus showed great potential for industrial application. This work also demonstrated a procedure on how to integrate the existing knowledge to engineer a strain with industrial application potential.


Assuntos
Butanóis/metabolismo , Cromossomos Bacterianos/genética , Escherichia coli , Genes Bacterianos , Engenharia Metabólica , Microrganismos Geneticamente Modificados , Escherichia coli/genética , Escherichia coli/metabolismo , Microrganismos Geneticamente Modificados/genética , Microrganismos Geneticamente Modificados/metabolismo
15.
Appl Environ Microbiol ; 83(11)2017 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-28363967

RESUMO

Pyruvate kinase is one of the three rate-limiting glycolytic enzymes that catalyze the last step of glycolysis, conversion of phosphoenolpyruvate (PEP) into pyruvate, which is associated with ATP generation. Two isozymes of pyruvate kinase, PykF and PykA, are identified in Escherichia coli PykF is considered important, whereas PykA has a less-defined role. Prior studies inactivated the pykA gene to increase the level of its substrate, PEP, and thereby increased the yield of end products derived from PEP. We were surprised when we found a pykA::Tn5 mutant in a screen for increased yield of an end product derived from pyruvate (n-butanol), suggesting that the role of PykA needs to be reexamined. We show that the pykA mutant exhibited elevated intracellular ATP levels, biomass concentrations, glucose consumption, and n-butanol production. We also discovered that the pykA mutant expresses higher levels of a presumed pyruvate transporter, YhjX, permitting the mutant to recapture and metabolize excreted pyruvate. Furthermore, we demonstrated that the nucleotide diphosphate kinase activity of PykA leads to negative regulation of the intracellular ATP levels. Taking the data together, we propose that inactivation of pykA can be considered a general strategy to enhance the production of pyruvate-derived metabolites under anaerobic conditions.IMPORTANCE This study showed that knocking out pykA significantly increased the intracellular ATP level and thus significantly increased the levels of glucose consumption, biomass formation, and pyruvate-derived product formation under anaerobic conditions. pykA was considered to be encoding a dispensable pyruvate kinase; here we show that pykA negatively regulates the anaerobic glycolysis rate through regulating the energy distribution. Thus, knocking out pykA can be used as a general strategy to increase the level of pyruvate-derived fermentative products.


Assuntos
Trifosfato de Adenosina/metabolismo , Proteínas de Escherichia coli/metabolismo , Escherichia coli/enzimologia , Regulação Enzimológica da Expressão Gênica , Piruvato Quinase/metabolismo , Anaerobiose , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Escherichia coli/genética , Regulação Bacteriana da Expressão Gênica , Glucose/metabolismo , Fosfoenolpiruvato/metabolismo , Piruvato Quinase/genética
16.
Reprod Fertil Dev ; 29(2): 374-382, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26293651

RESUMO

The present study assessed the effect of birthweight on reproductive performance, including a possible mechanism, in male pigs. Ten newborn male piglets, including five normal birthweight (NBW) piglets and five intra-uterine growth restricted (IUGR) piglets, were used in the study. All piglets were weaned on Day 28 and fed the same diet during the experiment (10 months). Average daily weight gain, feed intake and the feed conversion ratio were higher in NBW than IUGR piglets. Similarly, testis volume and the number of Leydig and Sertoli cells in the distal portion of the testes were higher in NBW than IUGR piglets (P<0.05). Semen volume (P<0.05) and the total number of spermatozoa per ejaculate (P=0.08) were lower in IUGR boars. Testosterone concentrations on Day 141 and prostaglandin E2 concentrations on Days 82 and 141 were higher in IUGR than NBW boars (P<0.05). The malondialdehyde content of seminal plasma was higher in IUGR boars, whereas sperm glutathione peroxidase activity was lower in IUGR versus NBW boars (P<0.05). Expression of DNA methyltransferase (Dnmt) genes Dnmt1, Dnmt3a, histone-lysine N-methyltransferase (Suv39h2), and lysine (K)-specific demethylase Kdm4a was upregulated in testes from IUGR boars. These findings suggest that growth restriction affects sperm production via reproductive organ development and epigenetic regulation.


Assuntos
Fertilidade/fisiologia , Retardo do Crescimento Fetal/metabolismo , Células Intersticiais do Testículo/citologia , Sêmen/metabolismo , Células de Sertoli/citologia , Animais , Contagem de Células , Dinoprostona/sangue , Retardo do Crescimento Fetal/fisiopatologia , Glutationa Peroxidase/metabolismo , Células Intersticiais do Testículo/metabolismo , Masculino , Malondialdeído/metabolismo , Células de Sertoli/metabolismo , Espermatozoides/metabolismo , Suínos , Testosterona/sangue
17.
Appl Microbiol Biotechnol ; 100(3): 1523-1529, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26572521

RESUMO

In typical acetone-butanol-ethanol (ABE) fermentation, acetone is the main by-product (50 % of butanol mass) of butanol production, resulting in a low yield of butanol. It is known that some Clostridium tetanomorphum strains are able to produce butanol without acetone in nature. Here, we described that C. tetanomorphum strain DSM665 can produce 4.16 g/L butanol and 4.98 g/L ethanol at pH 6.0, and 9.81 g/L butanol and 1.01 g/L ethanol when adding 1 mM methyl viologen. Butyrate and acetate could be reassimilated and no acetone was produced. Further analysis indicated that the activity of the acetate/butyrate:acetoacetyl-CoA transferase responsible for acetone production is lost in C. tetanomorphum DSM665. The genome of C. tetanomorphum DSM665 was sequenced and deposited in DDBJ, EMBL, and GenBank under the accession no. APJS00000000. Sequence analysis indicated that there are no typical genes (ctfA/B and adc) that are typically parts of an acetone synthesis pathway in C. tetanomorphum DSM665. This work provides new insights in the mechanism of clostridial butanol production and should prove useful for the design of a high-butanol-producing strain.


Assuntos
1-Butanol/metabolismo , Acetona/metabolismo , Proteínas de Bactérias/genética , Clostridium tetanomorphum/genética , Clostridium tetanomorphum/metabolismo , Proteínas de Bactérias/metabolismo , Clostridium tetanomorphum/crescimento & desenvolvimento , Fermentação , Genoma Bacteriano , Genômica , Dados de Sequência Molecular
18.
Chemistry ; 21(51): 18528-33, 2015 Dec 14.
Artigo em Inglês | MEDLINE | ID: mdl-26462445

RESUMO

A unique CdS/C@Fe3O4 nanoreactor was fabricated by the surface-imprinting technique, which effectively enhances the recyclability, stability, and selectivity for orientation recognition and photodegradation of ciprofloxacin in the binary mixed solution under visible-light irradiation. This work not only puts forward a novel design idea that develops the potential application value of CdS, but also provides a new approach for inhibiting its secondary pollution.


Assuntos
Compostos de Cádmio/química , Ciprofloxacina/química , Compostos Férricos/química , Sulfetos/química , Catálise , Fotólise , Reciclagem
19.
Inorg Chem ; 54(24): 11826-30, 2015 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-26616241

RESUMO

Wide-light absorption is important to semiconductors exploited in many applications such as photocatalysts, photovoltaic devices, and light-emitting diodes, which can effectively improve solar energy utilization. Especially for photocatalysts, the development and design of new semiconductors that harvest the whole-visible-light region (λ = 400-800 nm) is rarely reported, which is still a tremendous challenge up to now. Here we realize whole-visible-light absorption up to 900 nm for a semiconductor by means of construction of a mixed-valence Ag0.68V2O5, which results from an assistant effect of d-d transition. Ag0.68V2O5 serving as a photocatalyst obviously exhibits photoelectrochemical and photocatalytic properties. Our results provide a brand-new feasible design strategy to broaden the light absorption of semiconductors and highlight a route to further make the best use of the full solar spectrum.

20.
Phys Chem Chem Phys ; 17(2): 795-9, 2015 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-25407471

RESUMO

The prepared Ag2Ta4O11 photocatalyst exhibits durable activity for H2 production from water. We investigated the durability, inactivation and regeneration mechanism in depth. This work provides a new perspective and makes an important step for the research on Ag-based photocatalysts.

SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA