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1.
Sci Total Environ ; 912: 169135, 2024 Feb 20.
Artículo en Inglés | MEDLINE | ID: mdl-38070572

RESUMEN

Cr(VI) is a well-known toxic pollutant and its remediation has attracted great attention. It is important to continuously discover and explore new high-efficiency Cr(VI) reducing bacteria to further improve the efficiency of Cr(VI) pollution remediation. In this paper, metabolic mechanism of Cr(VI) reduction in a new highly efficient Cr(VI) reducing bacterium, Alicycliphilus denitrificans Ylb10, was investigated. The results showed that Ylb10 could tolerate and completely reduce 450 mg/L Cr(VI). Cr(VI) can be reduced in the intracellular compartment, membrane and the extracellular compartment, with the plasma membrane being the main active site for Cr(VI) reduction. With the addition of NADH, the reduction efficiency of cell membrane components for Cr(VI) increased 2.3-fold. The omics data analysis showed that sulfite reductase CysJ, thiosulfate dehydrogenase TsdA, nitrite reductase NrfA, nitric oxide reductase NorB, and quinone oxidoreductase ChrR play important roles in the reduction of Cr(VI), in the intracellular, and the extracellular compartment, and the membrane of Ylb10, and therefore Cr(VI) was reduced by the combined action of several reductases at these three locations.


Asunto(s)
Comamonadaceae , Restauración y Remediación Ambiental , Cromo/química , Biodegradación Ambiental , Oxidación-Reducción
2.
Am J Transl Res ; 15(8): 5347-5355, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37692957

RESUMEN

OBJECTIVE: To explore the effect of modified Guizhi plus Gegen decoction combined with the blade needle therapy on traditional Chinese medicine (TCM) syndromes, cervical curvature, and inflammatory factor levels in patients with cervical spondylotic radiculopathy. METHODS: In this retrospective study, 114 patients with cervical spondylotic radiculopathy who visited Pain Clinic, Hangzhou Fuyang Hospital of TCM Orthopedics and Traumatology from January 2020 to December 2022 were selected as the study subjects. According to different treatment methods, these patients were divided into an observation group (n=57, treated with blade needle therapy) and a control group (n=57, treated with modified Guizhi plus Gegen decoction combined with the blade needle therapy). Patients in both groups were treated for 3 courses. The treatment effects, TCM syndrome scores, cervical curvature, hemorheology indexes, inflammatory factors and adverse reactions were analyzed and compared between the two groups. RESULTS: The effective rate of patients in the observation group was 94.74%, which was significantly higher than 82.46% in the control group (P<0.05). After treatment, TCM syndrome scores, hemorheology indexes, and inflammatory factors levels in both of groups were significantly decreased in contrast to before treatment, while the cervical curvature was obviously increased. Compared with the control group, after the treatment, TCM syndrome scores, hemorheology indexes, inflammatory factors levels after treatment in the observation group were obviously lower, while the cervical curvature in the observation group being significantly increased (all P<0.05). No statistical differences were found for the incidence of adverse reactions between two the groups. CONCLUSION: Modified Guizhi plus Gegen decoction combined with the blade needle therapy effectively improved the TCM syndrome scores, restored the curvature of the cervical spine, improved the hemorheology of patients, inhibited the levels of inflammatory factors and it also has few adverse reactions, with a significant treatment effect in patients with cervical spondylotic radiculopathy.

3.
Materials (Basel) ; 16(8)2023 Apr 12.
Artículo en Inglés | MEDLINE | ID: mdl-37109877

RESUMEN

Considering that a jointed rock mass in a cold area is often affected by periodic freeze-thaw cycles and shear failure, definitions for the mesoscopic and macroscopic damage to a jointed rock mass under the coupling of freeze-thaw and shear are proposed, and the damage mechanism is verified according to experimental results. The results show that: (1) the jointed rock specimens increase macro-joints and meso-defects, the mechanical properties deteriorate significantly under freeze-thaw cycles, and the damage degree becomes more and more significant with the increases in freeze-thaw cycles and joint persistency. (2) When the number of freeze-thaw cycles is constant, the total damage variable value gradually increases with the increase in joint persistency. The damage variable difference in specimens with different persistency is distinct, which is gradually reduced in the later cycles, indicating a weakening influence of persistency on the total damage variable. (3) The shear resistance of non-persistent jointed rock mass in a cold area is determined by the coupling effect of meso-damage and frost heaving macro-damage. The coupling damage variable can accurately describe the damage variation law of jointed rock mass under freeze-thaw cycles and shear load.

4.
J Pediatr Hematol Oncol ; 45(2): 99-102, 2023 03 01.
Artículo en Inglés | MEDLINE | ID: mdl-36716245

RESUMEN

Anaplastic large cell lymphoma (ALCL) is a rare non-Hodgkin T-cell lymphoma characterized by a cluster of differentiation-30 positivity. Subtypes are characterized by positive or negative anaplastic lymphoma kinase (ALK) expression. ALCLs account for about 10% to 15% of all pediatric non-Hodgkin lymphomas and more than 90% of the cases are ALK-positive. We report a rare case of pediatric systemic ALK-negative ALCL with an atypical presentation as a painful breast mass. Despite the general benign features of most pediatric breast masses, it is important to consider malignant systemic diagnoses like the one reported here.


Asunto(s)
Mama , Linfoma Anaplásico de Células Grandes , Linfoma de Células T Periférico , Linfoma de Células T , Niño , Humanos , Linfoma Anaplásico de Células Grandes/patología , Proteínas Tirosina Quinasas Receptoras/metabolismo , Mama/patología
5.
Gastroenterol Res Pract ; 2022: 4313647, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36304789

RESUMEN

Background: Magnetically controlled capsule endoscopy has been shown to be feasible for the examination of gastric diseases. However, there may be problems, such as low image quality, incomplete esophageal observation, and capsule retention. We developed a novel magnetically controlled cable capsule endoscopy (MCCCE) system and evaluated its feasibility through in vitro experiments. Methods: Three experienced endoscopists performed MCCCE on the plastic stomach model and the excised porcine stomach model 5 times, respectively. Outcomes included handle ability, examination time, examination completion, and image quality. The examination completion was accessed by other two blinded endoscopists, and the image quality was compared with conventional gastroscopy (Olympus, GIF-290). Results: The performance of MCCCE in vitro experiments is excellent, with a mean operation time of 18.5 minutes in the plastic stomach model and 17.3 minutes in the excised porcine stomach model. The visualization rate of the gastric mucosa is >90% in the plastic stomach model and 75-90% in the excised porcine stomach model. The images of MCCCE are very clear, with good color resolution and no image distortion, which seem to be comparable to conventional gastroscopy. Conclusions: MCCCE system is feasible for gastric examination in vitro experiments, living animal experiments and human trials need to be further conducted.

6.
Surg Endosc ; 36(12): 9454-9461, 2022 12.
Artículo en Inglés | MEDLINE | ID: mdl-36112221

RESUMEN

BACKGROUND: Magnetically controlled capsule endoscopy (MCCE) has recently increasingly been used for gastric examination. However, the image quality and esophageal observation is suboptimal. We developed a novel wired transmission magnetically controlled capsule endoscopy (WT-MCCE) system and evaluated its feasibility through in vitro and in vivo experiments. METHODS: A plastic stomach model and a pathological upper gastrointestinal model were used to evaluate the performance of WT-MCCE in vitro experiments. Twice of examination in the two in vitro models by WT-MCCE were performed by 5 endoscopists who were experienced in performing wireless capsule endoscopy. The examination of traditional gastroscopy (Olympus, GIF-HQ290) in the pathological upper gastrointestinal model was set as the control. In vivo experiments were performed in a live canine model by 3 endoscopists, in which WT-MCCE was inserted with the assistance of gastroscopy. Measurements included maneuverability, examination time, visualization of gastric mucosa, image quality and diagnostic accuracy. RESULTS: WT-MCCE showed good performance in both in vitro and in vivo experiments with excellent visualization of mucosa (75-100%). The mean operation time is 17.6 ± 2.7 min, 22.3 ± 1.9 min and 29.3 ± 3.4 min in three models, respectively. In pathological upper gastrointestinal model, all lesions, including esophageal varices, one polyp, one foreign body, two gastric ulcers and one duodenal ulcer, were detected by both WT-MCCE and traditional gastroscopy by all endoscopists. For the observation of esophagus and stomach in the canine model, WT-MCCE also showed excellent maneuverability and good image quality. CONCLUSIONS: The novel WT-MCCE system performed well in evaluating upper gastrointestinal landmarks and lesions in two in vitro models, and showed good performance in a canine model. WT-MCCE may be potentially useful for diagnosis of esophageal and gastric diseases.


Asunto(s)
Endoscopía Capsular , Úlcera Gástrica , Tracto Gastrointestinal Superior , Perros , Animales , Endoscopía del Sistema Digestivo/métodos , Gastroscopía/métodos , Tracto Gastrointestinal Superior/diagnóstico por imagen
7.
Microbiol Resour Announc ; 11(9): e0056422, 2022 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-35993778

RESUMEN

The transcriptomes of Zymomonas mobilis 2032 were captured during the fermentation of ammonia fiber expansion (AFEX)-pretreated corn stover and switchgrass hydrolysates containing different concentrations of glucose and xylose. RNA samples were collected when Z. mobilis was fermenting glucose or xylose. Here, we present transcriptome sequencing (RNA-Seq) data obtained during separate phases of glucose or xylose consumption.

8.
Materials (Basel) ; 15(12)2022 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-35744287

RESUMEN

This study aims to quantify the shear surface morphology of jointed rock and its evolution under shearing, cyclic freezing, and thawing using the Gaussian filtering method. Gaussian filtering method enables the construction of the (large-scale) waviness surface and the (small-scale) unevenness surface of a digitized surface (created by laser scanning). Both waviness and unevenness surfaces are then quantified by roughness coefficient ratio (S) and degradation degrees of the waviness surface (Dw) and unevenness surface (Dr). These (microscopic) morphological parameters (S, Dw and Dr) are subsequently used to explain the development of the (macroscopic) shear strength of the jointed rocks on direct shear tests. The results indicate that compared with fresh jointed rocks, the freezing and thawing causes the potential shear surface asperities to be easier to damage and fail under shear load. Such damage is well represented by the significant decrease in Dw and Dr. On the other hand, with the increase of the freeze-thaw cycle (N), Dw increases while Dr reaches the maximum at an early stage of the cycle, where Dr > Dw. This difference reveals the underlying shear mechanism microscopically; that is, in the initial stage, the shear surface morphology is mainly dominated by the unevenness surface Dr, and then it is controlled by the waviness surface Dw during the freeze-thaw cycle.

9.
Biotechnol Biofuels Bioprod ; 15(1): 70, 2022 Jun 24.
Artículo en Inglés | MEDLINE | ID: mdl-35751080

RESUMEN

BACKGROUND: Lignocellulosic conversion residue (LCR) is the material remaining after deconstructed lignocellulosic biomass is subjected to microbial fermentation and treated to remove the biofuel. Technoeconomic analyses of biofuel refineries have shown that further microbial processing of this LCR into other bioproducts may help offset the costs of biofuel generation. Identifying organisms able to metabolize LCR is an important first step for harnessing the full chemical and economic potential of this material. In this study, we investigated the aerobic LCR utilization capabilities of 71 Streptomyces and 163 yeast species that could be engineered to produce valuable bioproducts. The LCR utilization by these individual microbes was compared to that of an aerobic mixed microbial consortium derived from a wastewater treatment plant as representative of a consortium with the highest potential for degrading the LCR components and a source of genetic material for future engineering efforts. RESULTS: We analyzed several batches of a model LCR by chemical oxygen demand (COD) and chromatography-based assays and determined that the major components of LCR were oligomeric and monomeric sugars and other organic compounds. Many of the Streptomyces and yeast species tested were able to grow in LCR, with some individual microbes capable of utilizing over 40% of the soluble COD. For comparison, the maximum total soluble COD utilized by the mixed microbial consortium was about 70%. This represents an upper limit on how much of the LCR could be valorized by engineered Streptomyces or yeasts into bioproducts. To investigate the utilization of specific components in LCR and have a defined media for future experiments, we developed a synthetic conversion residue (SynCR) to mimic our model LCR and used it to show lignocellulose-derived inhibitors (LDIs) had little effect on the ability of the Streptomyces species to metabolize SynCR. CONCLUSIONS: We found that LCR is rich in carbon sources for microbial utilization and has vitamins, minerals, amino acids and other trace metabolites necessary to support growth. Testing diverse collections of Streptomyces and yeast species confirmed that these microorganisms were capable of growth on LCR and revealed a phylogenetic correlation between those able to best utilize LCR. Identification and quantification of the components of LCR enabled us to develop a synthetic LCR (SynCR) that will be a useful tool for examining how individual components of LCR contribute to microbial growth and as a substrate for future engineering efforts to use these microorganisms to generate valuable bioproducts.

10.
Front Surg ; 9: 877040, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35586506

RESUMEN

Objective: To design and develop a disposable superfine catheter system for visual examination of bile and pancreatic ducts and predict its clinical application value. Methods: The superfine triple-lumen catheter and miniature photography technology were used to design and produce a disposable superfine catheter for visual examination of bile and pancreatic ducts, and animal experiments were conducted to compare said catheter and SpyGlass™. Results: The designed and developed disposable superfine catheter for visual examination of bile ducts with a diameter of 2.4 mm could enter the third-order and fourth-order bile ducts in the animal liver and also the gallbladder via the cystic duct for observation. The said catheter has a water injection rate of 0.8 mL/s, 0.16 megapixels, a resolution of 400 × 400, a depth of field of 0.3 to 20 mm, and a tilting up angle >90°. Conclusion: The new disposable catheter for visual examination of bile ducts has a superfine diameter, easier operation, and clearer imaging, and is expected to have a higher clinical practical value.

11.
Synth Syst Biotechnol ; 7(2): 738-749, 2022 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-35387233

RESUMEN

Metabolic engineering strategies have been successfully implemented to improve the production of isobutanol, a next-generation biofuel, in Saccharomyces cerevisiae. Here, we explore how two of these strategies, pathway re-localization and redox cofactor-balancing, affect the performance and physiology of isobutanol producing strains. We equipped yeast with isobutanol cassettes which had either a mitochondrial or cytosolic localized isobutanol pathway and used either a redox-imbalanced (NADPH-dependent) or redox-balanced (NADH-dependent) ketol-acid reductoisomerase enzyme. We then conducted transcriptomic, proteomic and metabolomic analyses to elucidate molecular differences between the engineered strains. Pathway localization had a large effect on isobutanol production with the strain expressing the mitochondrial-localized enzymes producing 3.8-fold more isobutanol than strains expressing the cytosolic enzymes. Cofactor-balancing did not improve isobutanol titers and instead the strain with the redox-imbalanced pathway produced 1.5-fold more isobutanol than the balanced version, albeit at low overall pathway flux. Functional genomic analyses suggested that the poor performances of the cytosolic pathway strains were in part due to a shortage in cytosolic Fe-S clusters, which are required cofactors for the dihydroxyacid dehydratase enzyme. We then demonstrated that this cofactor limitation may be partially recovered by disrupting iron homeostasis with a fra2 mutation, thereby increasing cellular iron levels. The resulting isobutanol titer of the fra2 null strain harboring a cytosolic-localized isobutanol pathway outperformed the strain with the mitochondrial-localized pathway by 1.3-fold, demonstrating that both localizations can support flux to isobutanol.

12.
Appl Opt ; 61(1): 188-195, 2022 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-35200818

RESUMEN

A theoretically designed rotating shadow mask is proposed to optimize the uniformity of a simple rotation system, which makes full use of the width of the coating chamber. This method can fabricate a large-aperture optical component, the diameter of which is more than half the width of the coating machine. The rotating shadow mask is applied to correct the film thickness uniformity near the center point of simple plane substrate. The factors influencing the effect of the rotating shadow mask are simulated and discussed. Then the shape of the rotating shadow mask is theoretically designed, and the uniformity within a corresponding radius is well corrected. After determining the shape of the rotating shadow mask, an additional fixed shadow mask is calculated and used to improve the uniformity of the entire substrate. Through the application of the two shadow masks together, uniformity about 99.5% is obtained in the diameter of 640 mm on a 1100 mm coating machine.

13.
Metab Eng ; 68: 119-130, 2021 11.
Artículo en Inglés | MEDLINE | ID: mdl-34592433

RESUMEN

Bottlenecks in the efficient conversion of xylose into cost-effective biofuels have limited the widespread use of plant lignocellulose as a renewable feedstock. The yeast Saccharomyces cerevisiae ferments glucose into ethanol with such high metabolic flux that it ferments high concentrations of glucose aerobically, a trait called the Crabtree/Warburg Effect. In contrast to glucose, most engineered S. cerevisiae strains do not ferment xylose at economically viable rates and yields, and they require respiration to achieve sufficient xylose metabolic flux and energy return for growth aerobically. Here, we evolved respiration-deficient S. cerevisiae strains that can grow on and ferment xylose to ethanol aerobically, a trait analogous to the Crabtree/Warburg Effect for glucose. Through genome sequence comparisons and directed engineering, we determined that duplications of genes encoding engineered xylose metabolism enzymes, as well as TKL1, a gene encoding a transketolase in the pentose phosphate pathway, were the causative genetic changes for the evolved phenotype. Reengineered duplications of these enzymes, in combination with deletion mutations in HOG1, ISU1, GRE3, and IRA2, increased the rates of aerobic and anaerobic xylose fermentation. Importantly, we found that these genetic modifications function in another genetic background and increase the rate and yield of xylose-to-ethanol conversion in industrially relevant switchgrass hydrolysate, indicating that these specific genetic modifications may enable the sustainable production of industrial biofuels from yeast. We propose a model for how key regulatory mutations prime yeast for aerobic xylose fermentation by lowering the threshold for overflow metabolism, allowing mutations to increase xylose flux and to redirect it into fermentation products.


Asunto(s)
Proteínas de Saccharomyces cerevisiae , Xilosa , Biocombustibles , Fermentación , Glucosa , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo
14.
Microvasc Res ; 136: 104150, 2021 07.
Artículo en Inglés | MEDLINE | ID: mdl-33647341

RESUMEN

Sickle cell disease (SCD) is a disorder with repetitive vaso-occlusive crises resulting in microvascular obstruction and tissue ischemia that may lead to multi-organ ischemia and dysfunction. Nailfold videocapillaroscopy (NFC) is an imaging technique utilized in clinical rheumatology to visualize capillaries located near the fingertip. To characterize NFC abnormalities in the setting of pediatric SCD, we performed NFC using a video capillaroscope on 8 digits in 44 stable SCD patients and 65 age matched healthy controls. Mean capillary number was lower (6.4 ± 1.3 vs 7.5 ± 1.8, p = 0.001) in the SCD group compared to controls. The percentage of dilated capillaries was similar (7.1 ± 8.3 vs. 5.9 ± 8.2, p = 0.4). The large majority of capillaries visualized in the SCD and control groups were normal capillary types per the EULAR definition, with a similar percentage of normal, nonspecific capillary morphologies and abnormal types. Regarding normal capillary sub-types, the SCD group and controls exhibited similar percentages of stereotype hairpin shapes, and tortuous or once or twice crossing type capillaries. On multivariate analyses, mean capillary number was independently associated with SCD after adjusting for age, body mass index, systolic blood pressure and gender. In conclusion, pediatric SCD is associated with lower capillary number but similar percentage of dilated capillaries and morphology on NFC. In our SCD cohort, capillary number was unrelated to our available markers of disease severity, including history of sickle crises, previous hospitalization for crises or Hemoglobin F levels.


Asunto(s)
Anemia de Células Falciformes/diagnóstico por imagen , Angioscopía Microscópica , Microvasos/diagnóstico por imagen , Uñas/irrigación sanguínea , Adolescente , Estudios de Casos y Controles , Niño , Preescolar , Estudios Transversales , Femenino , Humanos , Masculino , Densidad Microvascular , Valor Predictivo de las Pruebas
15.
Environ Sci Pollut Res Int ; 28(16): 19866-19877, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33410044

RESUMEN

Chromium (Cr) is one of the most widely used heavy metals in industrial processes, resulting in water and soil pollution that seriously threaten environmental safety. In this paper, we have directionally enriched a Cr(VI)-reducing bacterial community YEM001 from no-Cr(VI) polluted pond sedimental sludge by selectively growing it in Cr(VI)-containing media. This community could effectively reduce Cr(VI) in laboratory rich media containing different concentrations of Cr(VI), such as 61% reduction at 435 mg/L Cr(VI), 85% reduction at 355 mg/L Cr(VI), and complete reduction at 269 mg/L Cr(VI) in 93.5 h. It was also able to completely reduce 100 mg/L and 300 mg/L Cr(VI) in landfill leachate and natural sludge in 48 h, respectively. Optimal pH for Cr(VI) reduction of the YEM001 is between 7 and 8 and the best efficiency for Cr(VI) reduction occurs at 30 °C. Metagenomic data demonstrated that the YEM001 community was composed of multiple bacteria, including well-known Cr(VI)-reducing bacteria and non-Cr(VI)-reducing bacteria. Delftia, Comamonas, Alicycliphilus, Acidovorax, Bacillus, and Clostridioides account for 83% of total community abundance. The stability of the composition of the YEM001 community and its Cr(VI)-reducing activity allows for its application in bioremediation of environmental Cr(VI) pollution.


Asunto(s)
Metales Pesados , Microbiota , Biodegradación Ambiental , Cromo , Oxidación-Reducción
16.
Gastroenterol Res Pract ; 2021: 2716559, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-35003252

RESUMEN

In the past 20 years, several magnetically controlled capsule endoscopes (MCCE) have been developed for the evaluation of gastric lesions, including NaviCam (ANKON), MiroCam-Navi (Intromedic), Endocapsule MGCE (Olympus and Siemens), SMCE (JIFU), and FAMCE (Jinshan). Although limited to observing esophageal and duodenal lesions and lacking the ability of biopsy, MCCE has the advantages of comfort, safety, no anesthesia, no risk of cross-infection, and high acceptability. Several high-quality RCTs showed that the diagnostic accuracy of MCCE is comparable to the traditional gastroscopy. Due to the nonnecessity of anesthesia, MCCE may be more suitable for the elderly with obvious comorbidities as well as children. With more evidences accumulated and more innovative technologies developed, MCCE is expected to be an important tool for screening of early gastric cancer or the diagnosis of gastric diseases.

17.
mSystems ; 5(4)2020 Jul 21.
Artículo en Inglés | MEDLINE | ID: mdl-32694125

RESUMEN

Zymomonas mobilis is an ethanologenic alphaproteobacterium with promise for the industrial conversion of renewable plant biomass into fuels and chemical bioproducts. Limited functional annotation of the Z. mobilis genome is a current barrier to both fundamental studies of Z. mobilis and its development as a synthetic biology chassis. To gain insight, we collected sample-matched multiomics data, including RNA sequencing (RNA-seq), transcription start site (TSS) sequencing (TSS-seq), termination sequencing (term-seq), ribosome profiling, and label-free shotgun proteomic mass spectrometry, across different growth conditions and used these data to improve annotation and assign functional sites in the Z. mobilis genome. Proteomics and ribosome profiling informed revisions of protein-coding genes, which included 44 start codon changes and 42 added proteins. We developed statistical methods for annotating transcript 5' and 3' ends, enabling the identification of 3,940 TSSs and their corresponding promoters and 2,091 transcription termination sites, which were distinguished from RNA processing sites by the lack of an adjacent RNA 5' end. Our results revealed that Z. mobilis σA -35 and -10 promoter elements closely resemble canonical Escherichia coli -35 and -10 elements, with one notable exception: the Z. mobilis -10 element lacks the highly conserved -7 thymine observed in E. coli and other previously characterized σA promoters. The σA promoters of another alphaproteobacterium, Caulobacter crescentus, similarly lack the conservation of -7 thymine in their -10 elements. Our results anchor the development of Z. mobilis as a platform for synthetic biology and establish strategies for empirical genome annotation that can complement purely computational methods.IMPORTANCE Efforts to rationally engineer synthetic pathways in Zymomonas mobilis are impeded by a lack of knowledge and tools for predictable and quantitative programming of gene regulation at the transcriptional, posttranscriptional, and posttranslational levels. With the detailed functional characterization of the Z. mobilis genome presented in this work, we provide crucial knowledge for the development of synthetic genetic parts tailored to Z. mobilis This information is vital as researchers continue to develop Z. mobilis for synthetic biology applications. Our methods and statistical analyses also provide ways to rapidly advance the understanding of poorly characterized bacteria via empirical data that enable the experimental validation of sequence-based prediction for genome characterization and annotation.

18.
Metab Eng ; 61: 261-274, 2020 09.
Artículo en Inglés | MEDLINE | ID: mdl-32590077

RESUMEN

The microbial production of chemicals and fuels from plant biomass offers a sustainable alternative to fossilized carbon but requires high rates and yields of bioproduct synthesis. Z. mobilis is a promising chassis microbe due to its high glycolytic rate in anaerobic conditions that are favorable for large-scale production. However, diverting flux from its robust ethanol fermentation pathway to nonnative pathways remains a major engineering hurdle. To enable controlled, high-yield synthesis of bioproducts, we implemented a central-carbon metabolism control-valve strategy using regulated, ectopic expression of pyruvate decarboxylase (Pdc) and deletion of chromosomal pdc. Metabolomic and genetic analyses revealed that glycolytic intermediates and NADH accumulate when Pdc is depleted and that Pdc is essential for anaerobic growth of Z. mobilis. Aerobically, all flux can be redirected to a 2,3-butanediol pathway for which respiration maintains redox balance. Anaerobically, flux can be redirected to redox-balanced lactate or isobutanol pathways with ≥65% overall yield from glucose. This strategy provides a promising path for future metabolic engineering of Z. mobilis.


Asunto(s)
Butanoles/metabolismo , Butileno Glicoles/metabolismo , Carbono/metabolismo , Microorganismos Modificados Genéticamente , Zymomonas , Anaerobiosis , Glucosa/genética , Glucosa/metabolismo , Microorganismos Modificados Genéticamente/genética , Microorganismos Modificados Genéticamente/metabolismo , Zymomonas/genética , Zymomonas/metabolismo
19.
ChemSusChem ; 13(8): 1922, 2020 Apr 21.
Artículo en Inglés | MEDLINE | ID: mdl-32285625

RESUMEN

Invited for this month's cover is the research team from the D.O.E. Great Lake Bioenergy Research Center (GLBRC) at the University of Wisconsin-Madison. The cover image shows how a diverse team with expertise in many different fields works together in an integrated fashion to address complex problems. Only when the whole system, from field to the liquid fuels and co-products, is assessed, can we identify the key parameters needed to design an economically viable biorefinery-based economy. Cover art by Chelsea Mamott. The Full Paper itself is available at 10.1002/cssc.201903345.

20.
ChemSusChem ; 13(8): 2012-2024, 2020 Apr 21.
Artículo en Inglés | MEDLINE | ID: mdl-31984673

RESUMEN

The hydroxycinnamic acids p-coumaric acid (pCA) and ferulic acid (FA) add diversity to the portfolio of products produced by using grass-fed lignocellulosic biorefineries. The level of lignin-bound pCA in Zea mays was modified by the alteration of p-coumaroyl-CoA monolignol transferase expression. The biomass was processed in a lab-scale alkaline-pretreatment biorefinery process and the data were used for a baseline technoeconomic analysis to determine where to direct future research efforts to couple plant design to biomass utilization processes. It is concluded that future plant engineering efforts should focus on strategies that ramp up accumulation of one type of hydroxycinnamate (pCA or FA) predominantly and suppress that of the other. Technoeconomic analysis indicates that target extraction titers of one hydroxycinnamic acid need to be >50 g kg-1 biomass, at least five times higher than observed titers for the impure pCA/FA product mixture from wild-type maize. The technical challenge for process engineers is to develop a viable process that requires more than 80 % reduction of the isolation costs.

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