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Metabolic Engineering

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https://www.readbyqxmd.com/read/29024819/a-modular-metabolic-engineering-approach-for-the-production-of-1-2-propanediol-from-glycerol-by-saccharomyces-cerevisiae
#1
Zia-Ul Islam, Mathias Klein, Maximilian R Aßkamp, Anders S R Ødum, Elke Nevoigt
Compared to sugars, a major advantage of using glycerol as a feedstock for industrial bioprocesses is the fact that this molecule is more reduced than sugars. A compound whose biotechnological production might greatly profit from the substrate's higher reducing power is 1,2-propanediol (1,2-PDO). Here we present a novel metabolic engineering approach to produce 1,2-PDO from glycerol in S. cerevisiae. Apart from implementing the heterologous methylglyoxal (MG) pathway for 1,2-PDO formation from dihydroxyacetone phosphate (DHAP) and expressing a heterologous glycerol facilitator, the employed genetic modifications included the replacement of the native FAD-dependent glycerol catabolic pathway by the `DHA pathway´ for delivery of cytosolic NADH and the reduction of triosephosphate isomerase (TPI) activity for increased precursor (DHAP) supply...
October 9, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28993212/biocatalytic-one-pot-diterminal-oxidation-and-esterification-of-n-alkanes-for-production-of-%C3%AE-%C3%AF-diol-and-%C3%AE-%C3%AF-dicarboxylic-acid-esters
#2
Youri M van Nuland, Fons A de Vogel, Elinor L Scott, Gerrit Eggink, Ruud A Weusthuis
Direct and selective terminal oxidation of medium-chain n-alkanes is a major challenge in chemistry. Efforts to achieve this have so far resulted in low specificity and overoxidized products. Biocatalytic oxidation of medium-chain n-alkanes - with for example the alkane monooxygenase AlkB from P. putida GPo1- on the other hand is highly selective. However, it also results in overoxidation. Moreover, diterminal oxidation of medium-chain n-alkanes is inefficient. Hence, α,ω-bifunctional monomers are mostly produced from olefins using energy intensive, multi-step processes...
October 7, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28974378/advanced-water-splitting-for-green-hydrogen-gas-production-through-complete-oxidation-of-starch-by-in-vitro-metabolic-engineering
#3
Jae-Eung Kim, Eui-Jin Kim, Hui Chen, Chang-Hao Wu, Michael W W Adams, Y-H Percival Zhang
Starch is a natural energy storage compound and is hypothesized to be a high-energy density chemical compound or solar fuel. In contrast to industrial hydrolysis of starch to glucose, an alternative ATP-free phosphorylation of starch was designed to generate cost-effective glucose 6-phosphate by using five thermophilic enzymes (i.e., isoamylase, alpha-glucan phosphorylase, 4-α-glucanotransferase, phosphoglucomutase, and polyphosphate glucokinase). This enzymatic phosphorolysis is energetically advantageous because the energy of α-1,4-glycosidic bonds among anhydroglucose units is conserved in the form of phosphorylated glucose...
September 30, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28962875/tat-proteins-as-novel-thylakoid-membrane-anchors-organize-a-biosynthetic-pathway-in-chloroplasts-and-increase-product-yield-5-fold
#4
Maria Perestrello Ramos Henriques de Jesus, Agnieszka Zygadlo Nielsen, Silas Busck Mellor, Annemarie Matthes, Meike Burow, Colin Robinson, Poul Erik Jensen
Photosynthesis drives the production of ATP and NADPH, and acts as a source of carbon for primary metabolism. NADPH is also used in the production of many natural bioactive compounds. These are usually synthesized in low quantities and are often difficult to produce by chemical synthesis due to their complex structures. Some of the crucial enzymes catalyzing their biosynthesis are the cytochromes P450 (P450s) situated in the endoplasmic reticulum (ER), powered by electron transfers from NADPH. Dhurrin is a cyanogenic glucoside and its biosynthesis involves a dynamic metabolon formed by two P450s, a UDP-glucosyltransferase (UGT) and a P450 oxidoreductase (POR)...
September 28, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28951266/fast-growth-phenotype-of-e-coli-k-12-from-adaptive-laboratory-evolution-does-not-require-intracellular-flux-rewiring
#5
Christopher P Long, Jacqueline E Gonzalez, Adam M Feist, Bernhard O Palsson, Maciek R Antoniewicz
Adaptive laboratory evolution (ALE) is a widely-used method for improving the fitness of microorganisms in selected environmental conditions. It has been applied previously to Escherichia coli K-12 MG1655 during aerobic exponential growth on glucose minimal media, a frequently used model organism and growth condition, to probe the limits of E. coli growth rate and gain insights into fast growth phenotypes. Previous studies have described up to 1.6-fold increases in growth rate following ALE, and have identified key causal genetic mutations and changes in transcriptional patterns...
September 23, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28951265/synthetic-biology-approaches-to-establish-a-heterologous-production-system-for-coronatines
#6
Katja Gemperlein, Michael Hoffmann, Liujie Huo, Patrick Pilak, Lutz Petzke, Rolf Müller, Silke C Wenzel
Coronatine (COR) represents a phytotoxin produced by several pathovars of Pseudomonas syringae. It mediates multiple virulence activities by mimicking the plant stress hormone jasmonoyl-l-isoleucine. Structurally, COR consists of a bicyclic polyketide moiety, coronafacic acid (CFA), which is linked via an amide bond to an unusual ethylcyclopropyl amino acid moiety, coronamic acid (CMA). In our studies, we aimed at establishing and engineering of heterologous COR and CFA production platforms using P. putida KT2440 as host...
September 23, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28951189/cofactor-self-sufficient-whole-cell-biocatalysts-for-the-production-of-2-phenylethanol
#7
Pengchao Wang, Xinwei Yang, Baixue Lin, Jianzhong Huang, Yong Tao
The efficiency of biocatalysis is often affected by an insufficient supply and regeneration of cofactors and redox equivalents. To alleviate this shortcoming, a cofactor self-sufficient system was developed for enhanced production of 2-phenylethanol (2-PE) in E. coli. A "bridge" between the amino acid and its corresponding alcohol was designed in the system using glutamate dehydrogenase. By coupling glutamate dehydrogenase with transaminase and alcohol dehydrogenase, the cosubstrate (2-oxoglutarate) and redox equivalents (NAD(P)H) were regenerated simultaneously, so that no external cofactor or redox source was required...
September 22, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28951188/-13-c-metabolic-flux-analysis-identifies-limitations-to-increasing-specific-productivity-in-fed-batch-and-perfusion
#8
Neil Templeton, Sen Xu, David J Roush, Hao Chen
Industrial cell culture requires substantial energy to generate protein. The protein generated is not only the product of interest (IgG in this case), but also the protein associated with biomass. Here, (13)C-Metabolic Flux Analysis ((13)C-MFA) was utilized to compare the stationary phase of a fed-batch process to a perfusion process producing the same product by the same clone. The fed-batch process achieved significantly higher specific productivity, approximately 60% greater than the perfusion process. In spite of this, a general lack of difference was observed when globally comparing glycolysis, pentose phosphate pathway, and TCA cycle fluxes...
September 22, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28943461/the-importance-of-sourcing-enzymes-from-non-conventional-fungi-for-metabolic-engineering-and-biomass-breakdown
#9
REVIEW
Susanna Seppälä, St Elmo Wilken, Doriv Knop, Kevin V Solomon, Michelle A O'Malley
A wealth of fungal enzymes has been identified from nature, which continue to drive strain engineering and bioprocessing for a range of industries. However, while a number of clades have been investigated, the vast majority of the fungal kingdom remains unexplored for industrial applications. Here, we discuss selected classes of fungal enzymes that are currently in biotechnological use, and explore more basal, non-conventional fungi and their underexploited biomass-degrading mechanisms as promising agents in the transition towards a bio-based society...
September 21, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28943460/engineering-a-bacterial-platform-for-total-biosynthesis-of-caffeic-acid-derived-phenethyl-esters-and-amides
#10
Jian Wang, Monika Mahajani, Sheneika L Jackson, Yaping Yang, Mengyin Chen, Eric M Ferreira, Yuheng Lin, Yajun Yan
Caffeic acid has been widely recognized as a versatile pharmacophore for synthesis of new chemical entities, among which caffeic acid derived phenethyl esters and amides are the most extensively-investigated bioactive compounds with potential therapeutical applications. However, the natural biosynthetic routes for caffeic acid derived phenethyl esters or amides remain enigmatic, limiting their bio-based production. Herein, product-directed design of biosynthetic schemes allowed the development of thermodynamically favorable pathways for these compounds via acyltransferase (ATF) mediated trans-esterification...
September 21, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28939278/engineering-the-biocatalytic-selectivity-of-iridoid-production-in-saccharomyces-cerevisiae
#11
John M Billingsley, Anthony B DeNicola, Joyann S Barber, Man-Cheng Tang, Joe Horecka, Angela Chu, Neil K Garg, Yi Tang
Monoterpene indole alkaloids (MIAs) represent a structurally diverse, medicinally essential class of plant derived natural products. The universal MIA building block strictosidine was recently produced in the yeast Saccharomyces cerevisiae, setting the stage for optimization of microbial production. However, the irreversible reduction of pathway intermediates by yeast enzymes results in a non-recoverable loss of carbon, which has a strong negative impact on metabolic flux. In this study, we identified and engineered the determinants of biocatalytic selectivity which control flux towards the iridoid scaffold from which all MIAs are derived...
September 20, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28939277/enabling-the-synthesis-of-medium-chain-alkanes-and-1-alkenes-in-yeast
#12
Zhiwei Zhu, Yongjin J Zhou, Min-Kyoung Kang, Anastasia Krivoruchko, Nicolaas A Buijs, Jens Nielsen
Microbial synthesis of medium chain aliphatic hydrocarbons, attractive drop-in molecules to gasoline and jet fuels, is a promising way to reduce our reliance on petroleum-based fuels. In this study, we enabled the synthesis of straight chain hydrocarbons (C7-C13) by yeast Saccharomyces cerevisiae through engineering fatty acid synthases to control the chain length of fatty acids and introducing heterologous pathways for alkane or 1-alkene synthesis. We carried out enzyme engineering/screening of the fatty aldehyde deformylating oxygenase (ADO), and compartmentalization of the alkane biosynthesis pathway into peroxisomes to improve alkane production...
September 20, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28928052/directing-enzyme-devolution-for-biosynthesis-of-alkanols-and-1-n-alkanediols-from-natural-polyhydroxy-compounds
#13
Lu Dai, Fei Tao, Hongzhi Tang, Yali Guo, Yaling Shen, Ping Xu
Primordial enzymes are proposed to possess broad specificities. Through divergence and evolution, enzymes have been refined to exhibit specificity towards one reaction or substrate, and are thus commonly assumed as "specialists". However, some enzymes are "generalists" that catalyze a range of substrates and reactions. This property has been defined as enzyme promiscuity and is of great importance for the evolution of new functions. The promiscuities of two enzymes, namely glycerol dehydratase and diol dehydratase, were herein exploited for catalyzing long-chain polyols, including 1,2-butanediol, 1,2,4-butanetriol, erythritol, 1,2-pentanediol, 1,2,5-pentanetriol, and 1,2,6-hexanetriol...
September 18, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28918285/controlling-cell-volume-for-efficient-phb-production-by-halomonas
#14
Xiao-Ran Jiang, Zhi-Hao Yao, Guo-Qiang Chen
Bacterial morphology is decided by cytoskeleton protein MreB and cell division protein FtsZ encoded by essential genes mreB and ftsZ, respectively. Inactivating mreB and ftsZ lead to increasing cell sizes and cell lengths, respectively, yet seriously reduce cell growth ability. Here we develop a temperature-responsible plasmid expression system for compensated expression of relevant gene(s) in mreB or ftsZ disrupted recombinants H. campaniensis LS21, allowing mreB or ftsZ disrupted recombinants to grow normally at 30°C in a bioreactor for 12h so that a certain cell density can be reached, followed by 36h cell size expansions or cell shape elongations at elevated 37°C at which the mreB and ftsZ encoded plasmid pTKmf failed to replicate in the recombinants and thus lost themselves...
September 14, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28916461/metabolic-engineering-of-escherichia-coli-for-the-synthesis-of-the-quadripolymer-poly-glycolate-co-lactate-co-3-hydroxybutyrate-co-4-hydroxybutyrate-from-glucose
#15
Zheng-Jun Li, Kangjian Qiao, Xue-Mei Che, Gregory Stephanopoulos
Escherichia coli was metabolically engineered to effectively produce a series of biopolymers consisted of four types of monomers including glycolate, lactate, 3-hydroxybutyrate and 4-hydroxybutyrate from glucose as the carbon source. The biosynthetic route of novel quadripolymers was achieved by the overexpression of a range of homologous and heterologous enzymes including isocitrate lyase, isocitrate dehydrogenase kinase/phosphatase, glyoxylate/hydroxypyruvate reductase, propionyl-CoA transferase, β-ketothiolase, acetoacetyl-CoA reductase, succinate semialdehyde dehydrogenase, 4-hydroxybutyrate dehydrogenase, CoA transferase and PHA synthase...
September 13, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28890188/harnessing-the-respiration-machinery-for-high-yield-production-of-chemicals-in-metabolically-engineered-lactococcus-lactis
#16
Jianming Liu, Zhihao Wang, Vijayalakshmi Kandasamy, Sang Yup Lee, Christian Solem, Peter Ruhdal Jensen
When modifying the metabolism of living organisms with the aim of achieving biosynthesis of useful compounds, it is essential to ensure that it is possible to achieve overall redox balance. We propose a generalized strategy for this, based on fine-tuning of respiration. The strategy was applied on metabolically engineered Lactococcus lactis strains to optimize the production of acetoin and (R,R)-2,3-butanediol (R-BDO). In the absence of an external electron acceptor, a surplus of two NADH per acetoin molecule is produced...
September 8, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28867349/engineering-escherichia-coli-membrane-phospholipid-head-distribution-improves-tolerance-and-production-of-biorenewables
#17
Zaigao Tan, Pouyan Khakbaz, Yingxi Chen, Jeremy Lombardo, Jong Moon Yoon, Jacqueline V Shanks, Jeffery B Klauda, Laura R Jarboe
Economically competitive microbial production of biorenewable fuels and chemicals is often impeded by toxicity of the product to the microbe. Membrane damage is often identified as a major mechanism of this toxicity. Prior efforts to strengthen the microbial membrane by changing the phospholipid distribution have largely focused on the fatty acid tails. Herein, a novel strategy of phospholipid head engineering is demonstrated in Escherichia coli. Specifically, increasing the expression of phosphatidylserine synthase (+pssA) was found to significantly increase both the tolerance and production of octanoic acid, a representative membrane-damaging solvent...
September 1, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28864262/balanced-activation-of-ispg-and-isph-to-eliminate-mep-intermediate-accumulation-and-improve-isoprenoids-production-in-escherichia-coli
#18
Qingyan Li, Feiyu Fan, Xiang Gao, Chen Yang, Changhao Bi, Jinlei Tang, Tao Liu, Xueli Zhang
The MEP pathway genes were modulated to investigate whether there were new rate-limiting steps and toxic intermediates in this pathway. Activating IspG led to significant decrease of cell growth and β-carotene production. It was found that ispG overexpression led to accumulation of intermediate HMBPP, which seriously interfered with synthesis machinery of nucleotide and protein in Escherichia coli. Activation of the downstream enzyme IspH could solve HMBPP accumulation problem and eliminate the negative effects of ispG overexpression...
August 31, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28803913/synthetic-rescue-couples-nadph-generation-to-metabolite-overproduction-in-saccharomyces-cerevisiae
#19
Siavash Partow, Patrick B Hyland, Radhakrishnan Mahadevan
Engineering the redox cofactor metabolism is known to be a key challenge in developing a platform strain for biosynthesis of valuable products. Hence, general strategies for manipulation of co-factor metabolism in industrially relevant hosts are of significance. Here, we demonstrate an improvement in α-ketoglutarate (AKG) production in S. cerevisiae using a novel approach based on synthetic rescue. Here, we first perturb the cytosolic NADPH metabolism via deletion of glucose-6-phosphate dehydrogenase (ZWF1)...
August 10, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28800965/the-crispr-cas9-facilitated-multiplex-pathway-optimization-cfpo-technique-and-its-application-to-improve-the-escherichia-coli-xylose-utilization-pathway
#20
Xinna Zhu, Dongdong Zhao, Huanna Qiu, Feiyu Fan, Shuli Man, Changhao Bi, Xueli Zhang
One of the most important research subjects of metabolic engineering is the pursuit of balanced metabolic pathways, which requires the modulation of expression of many genes. However, simultaneously modulating multiple genes on the chromosome remains challenging in prokaryotic organisms, including the industrial workhorse - Escherichia coli. In this work, the CRISPR/Cas9-facilitated multiplex pathway optimization (CFPO) technique was developed to simultaneously modulate the expression of multiple genes on the chromosome...
August 9, 2017: Metabolic Engineering
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