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Metabolic Engineering

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https://www.readbyqxmd.com/read/29462662/a-balanced-atp-driving-force-module-for-enhancing-photosynthetic-biosynthesis-of-3-hydroxybutyrate-from-co-2
#1
Jason T Ku, Ethan I Lan
Using engineered photoautotrophic microorganisms for the direct chemical synthesis from CO2 is an attractive direction for both sustainability and CO2 mitigation. However, the behaviors of non-native metabolic pathways may be difficult to control due to the different intracellular contexts between natural and heterologous hosts. While most metabolic engineering efforts focus on strengthening driving forces in pathway design to favor biochemical production in these organisms, excessive driving force may be detrimental to product biosynthesis due to imbalanced cellular intermediate distribution...
February 17, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/29427605/a-mathematical-framework-for-yield-versus-rate-optimization-in-constraint-based-modeling-and-applications-in-metabolic-engineering
#2
Steffen Klamt, Stefan Müller, Georg Regensburger, Jürgen Zanghellini
BACKGROUND: The optimization of metabolic rates (as linear objective functions) represents the methodical core of flux-balance analysis techniques which have become a standard tool for the study of genome-scale metabolic models. Besides (growth and synthesis) rates, metabolic yields are key parameters for the characterization of biochemical transformation processes, especially in the context of biotechnological applications. However, yields are ratios of rates, and hence the optimization of yields (as nonlinear objective functions) under arbitrary linear constraints is not possible with current flux-balance analysis techniques...
February 7, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/29408291/model-assisted-metabolic-engineering-of-escherichia-coli-for-long-chain-alkane-and-alcohol-production
#3
Zia Fatma, Hassan Hartman, Mark G Poolman, David A Fell, Shireesh Srivastava, Tabinda Shakeel, Syed Shams Yazdani
Biologically-derived hydrocarbons are considered to have great potential as next-generation biofuels owing to the similarity of their chemical properties to contemporary diesel and jet fuels. However, the low yield of these hydrocarbons in biotechnological production is a major obstacle for commercialization. Several genetic and process engineering approaches have been adopted to increase the yield of hydrocarbon, but a model driven approach has not been implemented so far. Here, we applied a constraint-based metabolic modeling approach in which a variable demand for alkane biosynthesis was imposed, and co-varying reactions were considered as potential targets for further engineering of an E...
February 3, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/29330068/engineering-of-hydroxymandelate-synthases-and-the-aromatic-amino-pathway-enables-de-novo-biosynthesis-of-mandelic-and-4-hydroxymandelic-acid-with-saccharomyces-cerevisiae
#4
Mara Reifenrath, Eckhard Boles
Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose...
January 9, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/29289724/disruption-of-the-cell-wall-integrity-gene-ecm33-results-in-improved-fermentation-performance-by-wine-yeast
#5
Jin Zhang, Maria A Astorga, Jennifer M Gardner, Michelle E Walker, Paul R Grbin, Vladimir Jiranek
Limiting assimilable nitrogen restricts yeast's ability to successfully complete fermentation. In the absence of a commercially available deletion collection in a wine yeast background, a screening approach was applied to a transposon library in a wine yeast derivative to identify clones with superior fermentation performance. Eight candidate genes, when disrupted by Ty insertion, were identified as enabling yeast to efficiently complete a model oenological fermentation with limited nitrogen availability. Analogous single gene disruptions were subsequently constructed in the haploid wine yeast strain, C911D and the performance of these during fermentation was analysed...
December 28, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/29278749/thermodynamic-analysis-of-computed-pathways-integrated-into-the-metabolic-networks-of-e-coli-and-synechocystis-reveals-contrasting-expansion-potential
#6
Johannes Asplund-Samuelsson, Markus Janasch, Elton P Hudson
Introducing biosynthetic pathways into an organism is both reliant on and challenged by endogenous biochemistry. Here we compared the expansion potential of the metabolic network in the photoautotroph Synechocystis with that of the heterotroph E. coli using the novel workflow POPPY (Prospecting Optimal Pathways with PYthon). First, E. coli and Synechocystis metabolomic and fluxomic data were combined with metabolic models to identify thermodynamic constraints on metabolite concentrations (NET analysis). Then, thousands of automatically constructed pathways were placed within each network and subjected to a network-embedded variant of the max-min driving force analysis (NEM)...
December 23, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/29258965/tailored-carbon-partitioning-for-phototrophic-production-of-e-%C3%AE-bisabolene-from-the-green-microalga-chlamydomonas-reinhardtii
#7
Julian Wichmann, Thomas Baier, Eduard Wentnagel, Kyle J Lauersen, Olaf Kruse
Photosynthetic microbial hosts such as cyanobacteria and eukaryotic microalgae have recently emerged as alternative engineering platforms for the sustainable light-driven bio-production of terpenoids. Many desirable compounds with numerous applications can be produced in microorganisms by heterologous expression of terpene synthases. However, success of green microbial systems has been hampered by issues such as insufficient enzyme expression titers and low flux to desired terpenoid products from carbon fixed during photosynthesis...
December 16, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/29258964/engineered-monoculture-and-co-culture-of-methylotrophic-yeast-for-de-novo-production-of-monacolin-j-and-lovastatin-from-methanol
#8
Yiqi Liu, Xiaohu Tu, Qin Xu, Chenxiao Bai, Chuixing Kong, Qi Liu, Jiahui Yu, Qiangqiang Peng, Xiangshan Zhou, Yuanxing Zhang, Menghao Cai
As a promising one-carbon renewable substrate for industrial biotechnology, methanol has attracted much attention. However, engineering of microorganisms for industrial production of pharmaceuticals using a methanol substrate is still in infancy. In this study, the methylotrophic yeast Pichia pastoris was used to produce anti-hypercholesterolemia pharmaceuticals, lovastatin and its precursor monacolin J, from methanol. The biosynthetic pathways for monacolin J and lovastatin were first assembled and optimized in single strains using single copies of the relevant biosynthetic genes, and yields of 60...
December 16, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/29248755/construction-of-a-synthetic-metabolic-pathway-for-the-production-of-2-4-dihydroxybutyric-acid-from-homoserine
#9
Thomas Walther, Florence Calvayrac, Yoann Malbert, Ceren Alkim, Clémentine Dressaire, Hélène Cordier, Jean Marie François
2,4-dihydroxybutyrate (DHB) is a precursor for the chemical synthesis of the methionine analogue 2-hydroxy-4-(methylthio)butyrate. Since no annotated metabolic pathway exists for its microbial production from sugar, we have conceived a two-step synthetic metabolic pathway which converts the natural amino acid homoserine to DHB. The pathway proceeds through the homoserine transaminase-catalyzed deamination of homoserine to obtain 2-oxo-4-hydroxybutyrate (OHB), and continues with the reduction of OHB to DHB, which is catalyzed by an OHB reductase enzyme...
December 14, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/29247866/high-titer-production-of-lathyrane-diterpenoids-from-sugar-by-engineered-saccharomyces-cerevisiae
#10
Jeff Wong, Tristan de Rond, Leo d'Espaux, Cas van der Horst, Ishaan Dev, Leo Rios-Solis, James Kirby, Henrik Scheller, Jay Keasling
Euphorbiaceae are an important source of medically important diterpenoids, such as the anticancer drug ingenol-3-angelate and the antiretroviral drug prostratin. However, extraction from the genetically intractable natural producers is often limited by the small quantities produced, while the organic synthesis of terpene-derived drugs is challenging and similarly low-yielding. While transplanting the biosynthetic pathway into a heterologous host has proven successful for some drugs, it has been largely unsuccessful for diterpenoids due to their elaborate biosynthetic pathways and lack of genetic resources and tools for gene discovery...
December 13, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/29247865/bioproduction-of-a-betalain-color-palette-in-saccharomyces-cerevisiae
#11
Parbir S Grewal, Cyrus Modavi, Zachary N Russ, Nicholas C Harris, John E Dueber
Betalains are a family of natural pigments found exclusively in the plant order Caryophyllales. All members of this chemical family are biosynthesized through the common intermediate betalamic acid, which is capable of spontaneously condensing with various primary and secondary amines to produce betalains. Of particular interest is the red-violet betanin, most commonly obtained from Beta vulgaris (beet) as a natural food dye. We demonstrate the first complete microbial production of betanin in Saccharomyces cerevisiae from glucose, an early step towards a fermentation process enabling rapid, on-demand production of this natural dye...
December 13, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/29247864/microbial-synthesis-of-pyrogallol-using-genetically-engineered-escherichia-coli
#12
Jia Wang, Xiaolin Shen, Qipeng Yuan, Yajun Yan
Pyrogallol is a simple phenolic compound that serves as an attractive chemical with broad applications in food, agricultural, dyeing, printing, cosmetic, photography, chemical and pharmaceutical industries owing to its antioxidant, antibacterial, antiseptic, anticancer, oxygen-absorbing and strong reducing properties. Currently, pyrogallol is commercially produced by thermal decarboxylation of gallic acid under high temperature and pressure. However, this process is limited by the inaccessible raw material, the strict reaction conditions and the relatively low product yield...
December 13, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/29246517/enabling-the-valorization-of-guaiacol-based-lignin-integrated-chemical-and-biochemical-production-of-cis-cis-muconic-acid-using-metabolically-engineered-amycolatopsis-sp-atcc-39116
#13
Nadja Barton, Liliya Horbal, Sören Starck, Michael Kohlstedt, Andriy Luzhetskyy, Christoph Wittmann
Lignin is nature's second most abundant polymer and displays a largely unexploited renewable resource for value-added bio-production. None of the lignin-based fermentation processes so far managed to use guaiacol (2-methoxy phenol), the predominant aromatic monomer in depolymerized lignin. In this work, we describe metabolic engineering of Amycolatopsis sp. ATCC 39116 to produce cis,cis-muconic acid (MA), a precursor of recognized industrial value for commercial plastics, from guaiacol. The microbe utilized a very broad spectrum of lignin-based aromatics, such as catechol, guaiacol, phenol, toluene, p-coumarate, and benzoate, tolerated them in elevated amounts and even preferred them over sugars...
December 12, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/29242163/changes-of-dna-topology-affect-the-global-transcription-landscape-and-allow-rapid-growth-of-a-bacillus-subtilis-lacking-carbon-catabolite-repression
#14
Daniel R Reuß, Hermann Rath, Andrea Thürmer, Martin Benda, Rolf Daniel, Uwe Völker, Ulrike Mäder, Fabian M Commichau, Jörg Stülke
Bacteria are able to prioritize preferred carbon sources from complex mixtures. This is achieved by the regulatory phenomenon of carbon catabolite repression. To allow the simultaneous utilization of multiple carbon sources and to prevent the time-consuming adaptation to each individual nutrient in biotechnological applications, mutants lacking carbon catabolite repression can be used. However, such mutants often exhibit pleiotropic growth defects. We have isolated and characterized mutations that overcome the growth defect of Bacillus subtilis ccpA mutants lacking the major regulator of catabolite repression, in particular their glutamate auxotrophy...
December 11, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/29233745/retropath2-0-a-retrosynthesis-workflow-for-metabolic-engineers
#15
Baudoin Delépine, Thomas Duigou, Pablo Carbonell, Jean-Loup Faulon
Synthetic biology applied to industrial biotechnology is transforming the way we produce chemicals. However, despite advances in the scale and scope of metabolic engineering, the research and development process still remains costly. In order to expand the chemical repertoire for the production of next generation compounds, a major engineering biology effort is required in the development of novel design tools that target chemical diversity through rapid and predictable protocols. Addressing that goal involves retrosynthesis approaches that explore the chemical biosynthetic space...
December 9, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/29229581/balancing-cellular-redox-metabolism-in-microbial-electrosynthesis-and-electro-fermentation-a-chance-for-metabolic-engineering
#16
REVIEW
Frauke Kracke, Bin Lai, Shiqin Yu, Jens O Krömer
More and more microbes are discovered that are capable of extracellular electron transfer, a process in which they use external electrodes as electron donors or acceptors for metabolic reactions. This feature can be used to overcome cellular redox limitations and thus optimising microbial production. The technologies, termed microbial electrosynthesis and electro-fermentation, have the potential to open novel bio-electro production platforms from sustainable energy and carbon sources. However, the performance of reported systems is currently limited by low electron transport rates between microbes and electrodes and our limited ability for targeted engineering of these systems due to remaining knowledge gaps about the underlying fundamental processes...
December 8, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/29203223/expression-of-heterologous-non-oxidative-pentose-phosphate-pathway-from-bacillus-methanolicus-and-phosphoglucose-isomerase-deletion-improves-methanol-assimilation-and-metabolite-production-by-a-synthetic-escherichia-coli-methylotroph
#17
R Kyle Bennett, Jacqueline E Gonzalez, W Brian Whitaker, Maciek R Antoniewicz, Eleftherios T Papoutsakis
Synthetic methylotrophy aims to develop non-native methylotrophic microorganisms to utilize methane or methanol to produce chemicals and biofuels. We report two complimentary strategies to further engineer a previously engineered methylotrophic E. coli strain for improved methanol utilization. First, we demonstrate improved methanol assimilation in the presence of small amounts of yeast extract by expressing the non-oxidative pentose phosphate pathway (PPP) from Bacillus methanolicus. Second, we demonstrate improved co-utilization of methanol and glucose by deleting the phosphoglucose isomerase gene (pgi), which rerouted glucose carbon flux through the oxidative PPP...
December 5, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/29203222/methanol-assimilation-in-escherichia-coli-is-improved-by-co-utilization-of-threonine-and-deletion-of-leucine-responsive-regulatory-protein
#18
Jacqueline E Gonzalez, R Kyle Bennett, E Terry Papoutsakis, Maciek R Antoniewicz
Methane, the main component of natural gas, can be used to produce methanol which can be further converted to other valuable products. There is increasing interest in using biological systems for the production of fuels and chemicals from methanol, termed methylotrophy. In this work, we have examined methanol assimilation metabolism in a synthetic methylotrophic E. coli strain. Specifically, we applied 13C-tracers and evaluated 25 different co-substrates for methanol assimilation, including amino acids, sugars and organic acids...
December 2, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/29199104/reverse-engineering-the-cancer-metabolic-network-using-flux-analysis-to-understand-drivers-of-human-disease
#19
REVIEW
Mehmet G Badur, Christian M Metallo
Metabolic dysfunction has reemerged as an essential hallmark of tumorigenesis, and metabolic phenotypes are increasingly being integrated into pre-clinical models of disease. The complexity of these metabolic networks requires systems-level interrogation, and metabolic flux analysis (MFA) with stable isotope tracing present a suitable conceptual framework for such systems. Here we review efforts to elucidate mechanisms through which metabolism influences tumor growth and survival, with an emphasis on applications using stable isotope tracing and MFA...
November 30, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/29199103/diversion-of-the-long-chain-acyl-acp-pool-in-synechocystis-to-fatty-alcohols-through-crispri-repression-of-the-essential-phosphate-acyltransferase-plsx
#20
Danuta Kaczmarzyk, Ivana Cengic, Lun Yao, Elton P Hudson
Fatty alcohol production in Synechocystis sp. PCC 6803 was achieved through heterologous expression of the fatty acyl-CoA/ACP reductase Maqu2220 from the bacteria Marinobacter aquaeolei VT8 and the fatty acyl-ACP reductase DPW from the rice Oryza sativa. These platform strains became models for testing multiplex CRISPR-interference (CRISPRi) metabolic engineering strategies to both improve fatty alcohol production and to study membrane homeostasis. CRISPRi allowed partial repression of up to six genes simultaneously, each encoding enzymes of acyl-ACP-consuming pathways...
November 30, 2017: Metabolic Engineering
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