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Metabolic engineering

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https://www.readbyqxmd.com/read/28528488/enhanced-production-of-para-hydroxybenzoic-acid-by-genetically-engineered-saccharomyces-cerevisiae
#1
Nils J H Averesch, Alex Prima, Jens O Krömer
Saccharomyces cerevisiae is a popular organism for metabolic engineering; however, studies aiming at over-production of bio-replacement precursors for the chemical industry often fail to overcome proof-of-concept stage. When intending to show real industrial attractiveness, the challenge is twofold: formation of the target compound must be increased, while minimizing the formation of side and by-products to maximize titer, rate and yield. To tackle these, the metabolism of the organism, as well as the parameters of the process, need to be optimized...
May 20, 2017: Bioprocess and Biosystems Engineering
https://www.readbyqxmd.com/read/28527885/maintenance-of-atp-homeostasis-triggers-metabolic-shifts-in-gas-fermenting-acetogens
#2
Kaspar Valgepea, Renato de Souza Pinto Lemgruber, Kieran Meaghan, Robin William Palfreyman, Tanus Abdalla, Björn Daniel Heijstra, James Bruce Behrendorff, Ryan Tappel, Michael Köpke, Séan Dennis Simpson, Lars Keld Nielsen, Esteban Marcellin
Acetogens are promising cell factories for producing fuels and chemicals from waste feedstocks via gas fermentation, but quantitative characterization of carbon, energy, and redox metabolism is required to guide their rational metabolic engineering. Here, we explore acetogen gas fermentation using physiological, metabolomics, and transcriptomics data for Clostridium autoethanogenum steady-state chemostat cultures grown on syngas at various gas-liquid mass transfer rates. We observe that C. autoethanogenum shifts from acetate to ethanol production to maintain ATP homeostasis at higher biomass concentrations but reaches a limit at a molar acetate/ethanol ratio of ∼1...
May 16, 2017: Cell Systems
https://www.readbyqxmd.com/read/28527344/harnessing-plant-metabolic-diversity
#3
REVIEW
Charlie Owen, Nicola J Patron, Ancheng Huang, Anne Osbourn
Advances in DNA sequencing and synthesis technologies in the twenty-first century are now making it possible to build large-scale pipelines for engineering plant natural product pathways into heterologous production species using synthetic biology approaches. The ability to decode the chemical potential of plants by sequencing their transcriptomes and/or genomes and to then use this information as an instruction manual to make drugs and other high-value chemicals is opening up new routes to harness the vast chemical diversity of the Plant Kingdom...
May 17, 2017: Current Opinion in Chemical Biology
https://www.readbyqxmd.com/read/28522199/engineering-metabolic-pathways-in-escherichia-coli-for-constructing-a-microbial-chassis-for-biochemical-production
#4
REVIEW
Takuya Matsumoto, Tsutomu Tanaka, Akihiko Kondo
The present work reviews literature describing the re-design of the metabolic pathways of a microbial host using sophisticated genetic tools, yielding strains for producing value-added chemicals including fuels, building-block chemicals, pharmaceuticals, and derivatives. This work employed Escherichia coli, a well-studied microorganism that has been successfully engineered to produce various chemicals. E. coli has several advantages compared with other microorganisms, including robustness, and handling. To achieve efficient productivities of target compounds, an engineered E...
May 4, 2017: Bioresource Technology
https://www.readbyqxmd.com/read/28521055/tying-the-knot-occurrence-and-possible-significance-of-gene-fusions-in-plant-metabolism-and-beyond
#5
Jillian M Hagel, Peter J Facchini
Gene fusions have recently attracted attention especially in the field of plant specialized metabolism. The occurrence of a gene fusion, in which originally separate gene products are combined into a single polypeptide, often corresponds to the functional association of individual components within a single metabolic pathway. Examples include gene fusions implicated in benzylisoquinoline alkaloid (BIA), terpenoid, and amino acid biosynthetic pathways, in which distinct domains within a fusion catalyze consecutive, yet independent reactions...
May 18, 2017: Journal of Experimental Botany
https://www.readbyqxmd.com/read/28520394/in-vitro-metabolic-engineering-of-amorpha-4-11-diene-biosynthesis-at-enhanced-rate-and-specific-yield-of-production
#6
Xixian Chen, Congqiang Zhang, Ruiyang Zou, Gregory Stephanopoulos, Heng-Phon Too
In vitro metabolic engineering is an alternative approach to cell-based biosynthesis. It offers unprecedented flexibility for the study of biochemical pathways, thus, providing useful information for the rational design and assembly of reaction modules. Herein, we took the advantage of in vitro metabolic engineering to initially gain insight into the regulatory network of a reconstituted amorpha-4,11-diene (AD) synthetic pathway. Guided by lin-log approximation, we rapidly identified the hitherto unrecognized inhibition of adenosine triphosphate (ATP) on both farnesyl pyrophosphate synthase (FPPS) and amorpha-4,11-diene synthase (ADS)...
May 18, 2017: ACS Synthetic Biology
https://www.readbyqxmd.com/read/28518098/engineering-artificial-factors-to-specifically-manipulate-alternative-splicing-in-human-cells
#7
Huan-Huan Wei, Yuanlong Liu, Yang Wang, Qianyun Lu, Xuerong Yang, Jiefu Li, Zefeng Wang
The processing of most eukaryotic RNAs is mediated by RNA Binding Proteins (RBPs) with modular configurations, including an RNA recognition module, which specifically binds the pre-mRNA target and an effector domain. Previously, we have taken advantage of the unique RNA binding mode of the PUF domain in human Pumilio 1 to generate a programmable RNA binding scaffold, which was used to engineer various artificial RBPs to manipulate RNA metabolism. Here, a detailed protocol is described to construct Engineered Splicing Factors (ESFs) that are specifically designed to modulate the alternative splicing of target genes...
April 26, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28516163/engineered-heparins-as-new-anticoagulant-drugs
#8
REVIEW
Deepika Vaidyanathan, Asher Williams, Jonathan S Dordick, Mattheos A G Koffas, Robert J Linhardt
Heparin is an anionic polysaccharide that is widely used as a clinical anticoagulant. This glycosaminoglycan is prepared from animal tissues in metric ton quantities. Animal-sourced heparin is also widely used in the preparation of low molecular weight heparins that are gaining in popularity as a result of their improved pharmacological properties. The recent contamination of pharmaceutical heparin together with concerns about increasing demand for this life saving drug and the fragility of the heparin supply chain has led the scientific community to consider other potential sources for heparin...
March 2017: Bioengineering & Translational Medicine
https://www.readbyqxmd.com/read/28515315/the-c-terminal-peptide-of-aquifex-aeolicus-riboflavin-synthase-directs-encapsulation-of-native-and-foreign-guests-by-a-cage-forming-lumazine-synthase
#9
Yusuke Azuma, Reinhard Zschoche, Donald Hilvert
Encapsulation of specific enzymes in self-assembling protein cages is a hallmark of bacterial compartments that function as counterparts to eukaryotic organelles. The cage-forming enzyme lumazine synthase (LS) from Bacillus subtilis (BsLS), for example, encapsulates riboflavin synthase (BsRS), enabling channeling of lumazine from the site of its generation to the site of its conversion to vitamin B2. Elucidating the molecular mechanisms underlying the assembly of these supramolecular complexes could help inform new approaches for metabolic engineering, nanotechnology, and drug delivery...
May 17, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28513556/microscale-quantitative-analysis-of-polyhydroxybutyrate-in-prokaryotes-using-idms
#10
Mariana Itzel Velasco Alvarez, Angela Ten Pierick, Patricia T N van Dam, Reza Maleki Seifar, Mark C M van Loosdrecht, S Aljoscha Wahl
Poly(3-hydroxybutyrate) (PHB) is an interesting biopolymer for replacing petroleum-based plastics, its biological production is performed in natural and engineered microorganisms. Current metabolic engineering approaches rely on high-throughput strain construction and screening. Analytical procedures have to be compatible with the small scale and speed of these approaches. Here, we present a method based on isotope dilution mass spectrometry (IDMS) and propanolysis extraction of poly(3-hydroxybutyrate) from an Escherichia coli strain engineered for PHB production...
May 17, 2017: Metabolites
https://www.readbyqxmd.com/read/28509362/developing-polyamine-based-peptide-amphiphiles-with-tunable-morphology-and-physicochemical-properties
#11
Mehdi Bin Samad, Yashpal Singh Chhonker, Jacob I Contreras, Alec McCarthy, Megan M McClanahan, Daryl J Murry, Martin Conda-Sheridan
The ability to tune supramolecular properties such as size, morphology, or metabolic stability is of paramount importance in the field of supramolecular chemistry. Peptide amphiphiles (PAs) are a family of functional self-assembling biomaterials that have garnered widespread attention due to their broad applicability in medicine. PAs are generally comprised of an amino acid sequence connected to lipid tail(s) allowing them to self-assemble into supramolecular structures with diverse morphologies. Herein, this study describes the synthesis of a new class of polyamine-based "hybrid" PAs (PPAs) as novel self-assembling systems...
May 16, 2017: Macromolecular Bioscience
https://www.readbyqxmd.com/read/28508471/needleless-emulsion-electrospinning-for-the-regulated-delivery-of-susceptible-proteins
#12
Matej Buzgo, Eva Filova, Andrea Mickova Staffa, Michala Rampichova, Miroslav Doupnik, Karolina Vocetkova, Vera Lukasova, Radka Kolcun, David Lukas, Alois Necas, Evzen Amler
In the present work, we developed a novel needleless emulsion electrospinning technique which improves the production rate of the core/shell production process. The nanofibers are based on PCL as a continuous phase combined with a droplet phase based on Pluronic F-68 (PF-68). The PCL-PF-68 nanofibers show a time-regulated release of active molecules. Needleless emulsion electrospinning was used to encapsulate diverse set of compounds to the core phase (i.e. 5-DTAF-PF68, horseradish peroxidase (HRP), TRITC-dextran, insulin growth factor I (IGF-I), transforming growth factor beta (TGF-beta) and basic fibroblast growth factor (bFGF))...
May 16, 2017: Journal of Tissue Engineering and Regenerative Medicine
https://www.readbyqxmd.com/read/28503921/fluorescent-nanoparticles-with-tissue-dependent-affinity-for-live-zebrafish-imaging
#13
Deepak Kumar Khajuria, Vijay Bhooshan Kumar, David Karasik, Aharon Gedanken
Carbon-based nanodots (CDs) are widely investigated due to their low toxicity, outstanding water solubility and biocompatibility. Specifically, fluorescent CDs have attracted ever-increasing interest. However, so far, only a few studies have focused on assessing the fluorescence of nitrogen-doped CDs (N@CDs) during in-vivo exposure. Here, we describe a strategy for engineering economically viable non-toxic one-pot hydrothermal synthesized N@CDs. The low toxicity and suitability of the N@CDs for fluorescence imaging are validated using zebrafish (ZF) as a model...
May 15, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28503197/the-significance-of-proline-and-glutamate-on-butanol-chaotropic-stress-in-bacillus-subtilis-168
#14
Gumpanat Mahipant, Atchara Paemanee, Sittiruk Roytrakul, Junichi Kato, Alisa S Vangnai
BACKGROUND: Butanol is an intensively used industrial solvent and an attractive alternative biofuel, but the bioproduction suffers from its high toxicity. Among the native butanol producers and heterologous butanol-producing hosts, Bacillus subtilis 168 exhibited relatively higher butanol tolerance. Nevertheless, organic solvent tolerance mechanisms in Bacilli and Gram-positive bacteria have relatively less information. Thus, this study aimed to elucidate butanol stress responses that may involve in unique tolerance of B...
2017: Biotechnology for Biofuels
https://www.readbyqxmd.com/read/28502225/genomics-of-lactic-acid-bacteria-current-status-and-potential-applications
#15
Chongde Wu, Jun Huang, Rongqing Zhou
Lactic acid bacteria (LAB) are widely used for the production of a variety of foods and feed raw materials where they contribute to flavor and texture of the fermented products. In addition, specific LAB strains are considered as probiotic due to their health-promoting effects in consumers. Recently, the genome sequencing of LAB is booming and the increased amount of published genomics data brings unprecedented opportunity for us to reveal the important traits of LAB. This review describes the recent progress on LAB genomics and special emphasis is placed on understanding the industry-related physiological features based on genomics analysis...
August 2017: Critical Reviews in Microbiology
https://www.readbyqxmd.com/read/28501379/metabolic-impact-of-nutrient-starvation-in-mevalonate-producing-escherichia-coli
#16
Ami Masuda, Yoshihiro Toya, Hiroshi Shimizu
The aim of this work was to enhance mevalonate yield from glucose in Escherichia coli by essential nutrient starvations and to reveal these effects on the central carbon metabolism. Stationary phase culture without essential nutrients such as nitrogen, sulfur, and magnesium was evaluated using an engineered E. coli introducing mvaE and mvaS genes from Enterococcus faecalis. Sulfur starvation resulted in the highest mevalonate yield of 0.61C-molC-mol(-1) from glucose. The metabolic impacts of nutrient starvation were investigated by (13)C-metabolic flux analysis...
May 4, 2017: Bioresource Technology
https://www.readbyqxmd.com/read/28499817/use-of-cellnetanalyzer-in-biotechnology-and-metabolic-engineering
#17
Axel von Kamp, Sven Thiele, Oliver Hädicke, Steffen Klamt
Mathematical models of the cellular metabolism have become an essential tool for the optimization of biotechnological processes. They help to obtain a systemic understanding of the metabolic processes in the used microorganisms and to find suitable genetic modifications maximizing the production performance. In particular, methods of stoichiometric and constraint-based modeling are frequently used in the context of metabolic and bioprocess engineering. Since metabolic networks can be complex and comprise hundreds or even thousands of metabolites and reactions, dedicated software tools are required for an efficient analysis...
May 9, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28499254/improving-xylose-utilization-of-defatted-rice-bran-for-nisin-production-by-overexpression-of-a-xylose-transcriptional-regulator-in-lactococcus-lactis
#18
Jiaheng Liu, Zelin Ma, Hongji Zhu, Qinggele Caiyin, Dongmei Liang, Hao Wu, Xu Huang, Jianjun Qiao
Present investigation explores the potential of defatted rice bran (DRB) serving as sole carbon source and partial nitrogen source to support Lactococcus lactis growth and nisin production. To retain the nutrients in DRB, especially protein fractions, thermal pretreatment followed by enzymatic hydrolysis without washing step was applied for saccharification. A maximum of 45.64g reducing sugar mainly containing 30.26g glucose and 5.66g xylose from 100g DRB was attained in hydrolysates of DRB (HD). A novel strategy of xylR (xylose transcriptional regulator) overexpression followed by evolutionary engineering was proposed, which significantly increased the capacity of L...
April 22, 2017: Bioresource Technology
https://www.readbyqxmd.com/read/28498544/metabolic-engineering-of-cofactor-flavin-adenine-dinucleotide-fad-synthesis-and-regeneration-in-escherichia-coli-for-production-of-%C3%AE-keto-acids
#19
Ying Hou, Gazi Sakir Hossain, Jianghua Li, Hyun-Dong Shin, Guocheng Du, Jian Chen, Long Liu
Cofactor flavin adenine dinucleotide (FAD) plays a vital role in many FAD-dependent enzymatic reactions; therefore, how to efficiently accelerate FAD synthesis and regeneration is an important topic in biocatalysis and metabolic engineering. In this study, a system involving the synthesis pathway and regeneration of FAD was engineered in Escherichia coli to improve α-keto acid production-from the corresponding L-amino acids-catalyzed by FAD-dependent L-amino acid deaminase (L-AAD). First, key genes, ribH, ribC, and ribF, were overexpressed and fine-tuned for FAD synthesis...
May 12, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28498495/oleaginous-yeasts-promising-platforms-for-the-production-of-oleochemicals-and-biofuels
#20
José L Adrio
Oleaginous yeasts have a unique physiology that makes them the best suited hosts for the production of lipids, oleochemicals and diesel-like fuels. Their high lipogenesis, capability of growing on many different carbon sources (including lignocellulosic sugars), easy large-scale cultivation, and an increasing number of genetic tools are some of the advantages that have encouraged their use to develop sustainable processes. This mini-review summarizes the metabolic engineering strategies developed in oleaginous yeasts within the last two years to improve process metrics (titer, yield and productivity) for the production of lipids, free fatty acids, fatty acid-based chemicals (eg...
May 12, 2017: Biotechnology and Bioengineering
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