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Metabolic Engineering

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https://www.readbyqxmd.com/read/30439494/predicting-the-metabolic-capabilities-of-synechococcus-elongatus-pcc-7942-adapted-to-different-light-regimes
#1
Jared T Broddrick, David G Welkie, Denis Jallet, Susan S Golden, Graham Peers, Bernhard O Palsson
There is great interest in engineering photoautotrophic metabolism to generate bioproducts of societal importance. Despite the success in employing genome-scale modeling coupled with flux balance analysis to engineer heterotrophic metabolism, the lack of proper constraints necessary to generate biologically realistic predictions has hindered broad application of this methodology to phototrophic metabolism. Here we describe a methodology for constraining genome-scale models of photoautotrophy in the cyanobacteria Synechococcus elongatus PCC 7942...
November 12, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30391511/direct-cell-to-cell-exchange-of-matter-in-a-synthetic-clostridium-syntrophy-enables-co-2-fixation-superior-metabolite-yields-and-an-expanded-metabolic-space
#2
Kamil Charubin, Eleftherios Terry Papoutsakis
In microbial fermentations, at least 33% of the sugar-substrate carbon is lost as CO2 during pyruvate decarboxylation to acetyl-CoA, with the corresponding electrons lost in the form of H2 . Previous attempts to reduce this carbon and electron loss focused on engineering of a single organism. In nature, most microorganisms live in complex communities where syntrophic interactions result in superior resource utilization. Here, we show that a synthetic syntrophy consisting of the solventogen Clostridium acetobutylicum, which converts simple and complex carbohydrates into a variety of chemicals, and the acetogen C...
November 1, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30389613/upgrade-of-wood-sugar-d-xylose-to-a-value-added-nutraceutical-by-in-vitro-metabolic-engineering
#3
Kun Cheng, Wenming Zheng, Hongge Chen, Yi-Heng P Job Zhang
The upgrade of D-xylose, the most abundant pentose, to value-added biochemicals is economically important to next-generation biorefineries. myo-Inositol, as vitamin B8, has a six-carbon carbon-carbon ring. Here we designed an in vitro artificial NAD(P)-free 12-enzyme pathway that can effectively convert the five-carbon xylose to inositol involving xylose phosphorylation, carbon-carbon (C-C) rearrangement, C-C bond circulation, and dephosphorylation. The reaction conditions catalyzed by all thermostable enzymes from hyperthermophilic microorganisms Thermus thermophiles, Thermotoga maritima, and Archaeoglobus fulgidus were optimized in reaction temperature, buffer type and concentration, enzyme composition, Mg2+ concentration, and fed-batch addition of ATP...
October 31, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30389612/increased-sesqui-and-triterpene-production-by-co-expression-of-hmg-coa-reductase-and-biotin-carboxyl-carrier-protein-in-tobacco-nicotiana-benthamiana
#4
Ah-Reum Lee, Moonhyuk Kown, Min-Kyoung Kang, Jeonghan Kim, Soo-Un Kim, Dae-Kyun Ro
Terpenoids are the most diverse natural products with many industrial applications and are all synthesized from simple precursors, isopentenyl diphosphate (IPP) and its isomer dimethylallyl diphosphate (DMAPP). In plants, IPP is synthesized by two distinct metabolic pathways - cytosolic mevalonate (MVA) pathway for C15 sesquiterpene and C30 triterpene, and plastidic methylerythritol phosphate (MEP) pathway for C10 monoterpene and C20 diterpene. A number of studies have altered the metabolic gene expressions in either the MVA or MEP pathway to increase terpene production; however, it remains unknown if the alteration of the acetyl-CoA pool in plastid fatty acid biosynthesis can influence terpenoid flux...
October 30, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30393203/rewiring-metabolic-network-by-chemical-modulator-based-laboratory-evolution-doubles-lipid-production-in-crypthecodinium-cohnii
#5
Jinjin Diao, Xinyu Song, Jinyu Cui, Liangsen Liu, Mengliang Shi, Fangzhong Wang, Weiwen Zhang
Dietary omega-3 long-chain polyunsaturated fatty acids docosahexaenoic acid (DHA, C22:6) can be synthesized in microalgae Crypthecodinium cohnii; however, its productivity is still low. Here, we established a new protocol termed as "chemical modulator based adaptive laboratory evolution" (CM-ALE) to enhance lipid and DHA productivity in C. cohnii. First, ACCase inhibitor sethoxydim based CM-ALE was applied to redirect carbon equivalents from starch to lipid. Second, CM-ALE using growth modulator sesamol as selection pressure was conducted to relive negative effects of sesamol on lipid biosynthesis in C...
October 26, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30367967/expanding-biological-applications-using-cell-free-metabolic-engineering-an-overview
#6
REVIEW
James R Swartz
Expanding the concept of cell-free biology, implemented both with purified components and crude extracts, is continuing to deepen our appreciation of biological fundamentals while enlarging the range of applications. We are no longer intimidated by the complexity of crude extracts and complicated reaction systems with hundreds of active components, and, instead, coordinately activate and inactivate metabolic processes to focus and expand the capabilities of natural biological processes. This, in turn, dramatically increases the range of benefits offered by new products, both natural and supernatural, that were previously infeasible and/or unimaginable...
October 24, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30343048/synthetic-redesign-of-central-carbon-and-redox-metabolism-for-high-yield-production-of-n-acetylglucosamine-in-bacillus-subtilis
#7
Yang Gu, Xueqin Lv, Yanfeng Liu, Jianghua Li, Guocheng Du, Jian Chen, Ledesma-Amaro Rodrigo, Long Liu
One of the primary goals of microbial metabolic engineering is to achieve high titer, yield and productivity (TYP) of engineered strains. This TYP index requires optimized carbon flux toward desired molecule with minimal by-product formation. De novo redesign of central carbon and redox metabolism holds great promise to alleviate pathway bottleneck and improve carbon and energy utilization efficiency. The engineered strain, with the overexpression or deletion of multiple genes, typically can't meet the TYP index, due to overflow of central carbon and redox metabolism that compromise the final yield, despite a high titer or productivity might be achieved...
October 19, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30343047/efficient-production-of-3-hydroxypropionate-from-fatty-acids-feedstock-in-escherichia-coli
#8
Bo Liu, Shuman Xiang, Guang Zhao, Bojun Wang, Yanhe Ma, Weifeng Liu, Yong Tao
The production of chemicals from renewable biomass resources is usually limited by factors including high-cost processes and low efficiency of biosynthetic pathways. Fatty acids (FAs) are an ideal alternative biomass. Their advantages include high-efficiently producing acetyl-CoA and reducing power, coupling chemical production with CO2 fixation, and the fact that they are readily obtained from inexpensive feedstocks. The important platform chemical 3-hydroxypropionate (3HP) can be produced from FAs as the feedstock with a theoretical yield of 2...
October 19, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30339834/identification-of-a-novel-cytochrome-p450-enzyme-that-catalyzes-the-c-2%C3%AE-hydroxylation-of-pentacyclic-triterpenoids-and-its-application-in-yeast-cell-factories
#9
Zhubo Dai, Yun Liu, Zhoutong Sun, Dong Wang, Ge Qu, Xiaolin Ma, Feiyu Fan, Lili Zhang, Shoulian Li, Xueli Zhang
C-2α hydroxylated triterpenoids are a large class of plant secondary metabolites. These compounds, such as maslinic, corosolic and alphitolic acid, have important biological activities against HIV, cancer and diabetes. However, the biosynthesis pathways of these compounds have not been completely elucidated. Specifically, the cytochrome P450 (CYP) enzyme responsible for C-2α hydroxylation was unknown. In this study, a novel CYP enzyme that catalyzes C-2α hydroxylation was identified in Crataegus pinnatifida (Hawthorn) using a metabolic engineering platform...
October 16, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30268818/biosynthesis-of-monoethylene-glycol-in-saccharomyces-cerevisiae-utilizing-native-glycolytic-enzymes
#10
Boonsom Uranukul, Benjamin M Woolston, Gerald R Fink, Gregory Stephanopoulos
Monoethylene glycol (MEG) is an important commodity chemical with applications in numerous industrial processes, primarily in the manufacture of polyethylene terephthalate (PET) polyester used in packaging applications. In the drive towards a sustainable chemical industry, bio-based production of MEG from renewable biomass has attracted growing interest. Recent attempts for bio-based MEG production have investigated metabolic network modifications in Escherichia coli, specifically rewiring the xylose assimilation pathways for the synthesis of MEG...
September 27, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30240601/secondary-metabolites-overproduction-through-transcriptional-gene-cluster-refactoring
#11
Liliya Horbal, Filipe Marques, Suvd Nadmid, Marta V Mendes, Andriy Luzhetskyy
We present a random rational approach enabling the construction of overproducing strains in two steps. The approach first involves creating a library of clusters of interest, in which native promoters are substituted with randomly generated constitutive synthetic promoters, and then expressing this library in an appropriate host strain. This strategy is fast, easy to use, accounts for the architecture of a cluster and completely decouples the expression of a gene cluster from complex native regulatory networks...
September 18, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30227251/application-of-a-curated-genome-scale-metabolic-model-of-cho-dg44-to-an-industrial-fed-batch-process
#12
Cyrielle Calmels, Andréa McCann, Laetitia Malphettes, Mikael Rørdam Andersen
CHO cells have become the favorite expression system for large scale production of complex biopharmaceuticals. However, industrial strategies for upstream process development are based on empirical results, due to a lack of fundamental understanding of intracellular activities. Genome scale models of CHO cells have been reconstructed to provide an economical way of analyzing and interpreting large-omics datasets, since they add cellular context to the data. Here the most recently available CHO-DG44 genome-scale specific model was manually curated and tailored to the metabolic profile of cell lines used for industrial protein production, by modifying 601 reactions...
September 15, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30223026/high-production-of-4-hydroxyisoleucine-in-corynebacterium-glutamicum-by-multistep-metabolic-engineering
#13
Chenglin Zhang, Yanjun Li, Jie Ma, Yuan Liu, Jilong He, Yingzi Li, Fuzhou Zhu, Jing Meng, Junjie Zhan, Zhixiang Li, Lei Zhao, Qian Ma, Xiaoguang Fan, Qingyang Xu, Xixian Xie, Ning Chen
4-Hydroxyisoleucine (4-HIL) exhibits a unique glucose-dependent insulinotropic activity and is a promising candidate for the treatment of diabetes. Direct fermentation of 4-HIL has been recently studied; however, the expected titre and yield were not achieved. In this study, we initially developed a pathway for the synthesis of 4-HIL in an L-isoleucine producer, C. glutamicum YI, but insufficient supply of α-ketoglutarate was a bottleneck for a strong production. Six genes involved in oxaloacetate and α-ketoglutarate branches were overexpressed or deleted, which increased the production of 4-HIL to 5...
September 14, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30219528/engineering-nadh-nad-ratio-in-halomonas-bluephagenesis-for-enhanced-production-of-polyhydroxyalkanoates-pha
#14
Chen Ling, Guan-Qing Qiao, Bo-Wen Shuai, Karel Olavarria, Jin Yin, Rui-Juan Xiang, Kun-Nan Song, Yun-Hao Shen, Yingying Guo, Guo-Qiang Chen
Halomonas bluephagenesis has been developed as a platform strain for the next generation industrial biotechnology (NGIB) with advantages of resistances to microbial contamination and high cell density growth (HCD), especially for production of polyhydroxyalkanoates (PHA) including poly(3-hydroxybutyrate) (PHB), poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P34HB) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV). However, little is known about the mechanism behind PHA accumulation under oxygen limitation...
September 13, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30218716/characterization-of-the-clostridium-thermocellum-adhe-nfnab-ferredoxin-and-pfor-proteins-for-their-ability-to-support-high-titer-ethanol-production-in-thermoanaerobacterium-saccharolyticum
#15
Jingxuan Cui, Daniel G Olson, Lee R Lynd
The thermophilic anaerobes Thermoanaerobacterium saccharolyticum and Clostridium thermocellum are good candidates for lignocellulosic ethanol production. T. saccharolyticum has been successfully engineered to produce ethanol at high titer (70 g/L). The maximum ethanol titer of engineered strains of C. thermocellum is only 25 g/L. We hypothesize that one or more of the enzymes in the ethanol production pathway in C. thermocellum is not adequate for ethanol production at high titer. In this study, we focused on the enzymes responsible for the part of the ethanol production pathway from pyruvate to ethanol...
September 12, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30213650/engineering-saccharomyces-cerevisiae-for-production-of-simvastatin
#16
Carly M Bond, Yi Tang
Simvastatin is a semisynthetic cholesterol-lowering medication and one of the top-selling statins in the world. Currently, industrial production of simvastatin acid (SVA) is a multistep process starting from the natural product lovastatin. For this reason, there is significant interest in direct production of simvastatin from a microbial host. In this study, six heterologous biosynthetic genes were introduced into Saccharomyces cerevisiae and the acyl-donor dimethylbutyryl-S-methyl mercaptopropionate (DMB-SMMP) was added, resulting in initial production of 0...
September 10, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30201314/multiobjective-strain-design-a-framework-for-modular-cell-engineering
#17
Sergio Garcia, Cong T Trinh
Diversity of cellular metabolism can be harnessed to produce a large space of molecules. However, development of optimal strains with high product titers, rates, and yields required for industrial production is laborious and expensive. To accelerate the strain engineering process, we have recently introduced a modular cell design concept that enables rapid generation of optimal production strains by systematically assembling a modular cell with an exchangeable production module(s) to produce target molecules efficiently...
September 7, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30196100/generation-of-a-cluster-free-streptomyces-albus-chassis-strains-for-improved-heterologous-expression-of-secondary-metabolite-clusters
#18
Maksym Myronovskyi, Birgit Rosenkränzer, Suvd Nadmid, Petar Pujic, Philippe Normand, Andriy Luzhetskyy
Natural products are a rich source of potential drugs for many applications. Discovery of natural products through the activation of cryptic gene clusters encoding their biosynthetic pathways, engineering of those biosynthetic pathways and optimization of production yields often rely on the expression of these gene clusters in suitable heterologous host strains. Streptomyces albus J1074 provides high success rates of heterologous cluster expression with high levels of metabolite production, rapid growth and amenability to genetic manipulations...
September 6, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30195009/engineering-a-spermidine-biosynthetic-pathway-in-clostridium-thermocellum-results-in-increased-resistance-to-furans-and-increased-ethanol-production
#19
Sun-Ki Kim, Janet Westpheling
Polyamines are low molecular weight aliphatic nitrogen compounds found ubiquitously in microorganisms, plants, and animals. Spermidine is a common polyamine that plays a role in stabilizing chromatin, DNA replication, transcription, translation, as well as the regulation of cell growth and apoptosis in eukaryotes. Amines are also associated with defense to a number of environmental stresses including elevated temperature and have been shown to be involved in tolerance to fermentation inhibitors such as furan derivatives and acetic acid in Saccharomyces cerevisiae...
September 5, 2018: Metabolic Engineering
https://www.readbyqxmd.com/read/30189293/metabolic-engineering-of-escherichia-coli-for-high-yield-uridine-production
#20
Heyun Wu, Yanjun Li, Qian Ma, Qiang Li, Zifan Jia, Bo Yang, Qingyang Xu, Xiaoguang Fan, Chenglin Zhang, Ning Chen, Xixian Xie
Uridine is a kind of pyrimidine nucleoside that has been widely applied in the pharmaceutical industry. Although microbial fermentation is a promising method for industrial production of uridine, an efficient microbial cell factory is still lacking. In this study, we constructed a metabolically engineered Escherichia coli capable of high-yield uridine production. First, we developed a CRISPR/Cas9-mediated chromosomal integration strategy to integrate large DNA into the E. coli chromosome, and a 9.7 kb DNA fragment including eight genes in the pyrimidine operon of Bacillus subtilis F126 was integrated into the yghX locus of E...
September 4, 2018: Metabolic Engineering
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