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Microbial Cell Factories

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https://www.readbyqxmd.com/read/28103877/rhodococcus-erythropolis-as-a-host-for-expression-secretion-and-glycosylation-of-mycobacterium-tuberculosis-proteins
#1
Antonio J Vallecillo, Cristina Parada, Pedro Morales, Clara Espitia
BACKGROUND: Glycosylation is one of the most abundant posttranslational polypeptide chain modification in nature. Although carbohydrate modification of protein antigens from many microbial pathogens constitutes important components of B cell epitopes, the role in T cell immunity is not completely understood. There is growing evidence about the importance of these modifications in host bacteria interactions in tuberculosis. It is known, that the sugars present in some Mycobacterium tuberculosis glycoproteins play an important role in both humoral and cellular immune response against the pathogen...
January 19, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28100236/human-%C3%AE-defensin-2-production-from-s-cerevisiae-using-the-repressible-met17-promoter
#2
Thea S B Møller, Joanna Hay, Malcolm J Saxton, Karen Bunting, Evamaria I Petersen, Søren Kjærulff, Christopher J A Finnis
BACKGROUND: Baker's yeast Saccharomyces cerevisiae is a proven host for the commercial production of recombinant biopharmaceutical proteins. For the manufacture of heterologous proteins with activities deleterious to the host it can be desirable to minimise production during the growth phase and induce production late in the exponential phase. Protein expression by regulated promoter systems offers the possibility of improving productivity in this way by separating the recombinant protein production phase from the yeast growth phase...
January 18, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28095880/high-throughput-expression-of-animal-venom-toxins-in-escherichia-coli-to-generate-a-large-library-of-oxidized-disulphide-reticulated-peptides-for-drug-discovery
#3
Jeremy Turchetto, Ana Filipa Sequeira, Laurie Ramond, Fanny Peysson, Joana L A Brás, Natalie J Saez, Yoan Duhoo, Marilyne Blémont, Catarina I P D Guerreiro, Loic Quinton, Edwin De Pauw, Nicolas Gilles, Hervé Darbon, Carlos M G A Fontes, Renaud Vincentelli
BACKGROUND: Animal venoms are complex molecular cocktails containing a wide range of biologically active disulphide-reticulated peptides that target, with high selectivity and efficacy, a variety of membrane receptors. Disulphide-reticulated peptides have evolved to display improved specificity, low immunogenicity and to show much higher resistance to degradation than linear peptides. These properties make venom peptides attractive candidates for drug development. However, recombinant expression of reticulated peptides containing disulphide bonds is challenging, especially when associated with the production of large libraries of bioactive molecules for drug screening...
January 17, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28095865/engineered-biosynthesis-of-milbemycins-in-the-avermectin-high-producing-strain-streptomyces-avermitilis
#4
Myoun-Su Kim, Wan-Je Cho, Myoung Chong Song, Seong-Whan Park, Kaeun Kim, Eunji Kim, Naryeong Lee, Sang-Jip Nam, Ki-Hoon Oh, Yeo Joon Yoon
BACKGROUND: Milbemycins, produced from Streptomyces hygroscopicus subsp. aureolacrimosus and Streptomyces bingchenggensis, are 16-membered macrolides that share structural similarity with avermectin produced from Streptomyces avermitilis. Milbemycins possess strong acaricidal, insecticidal, and anthelmintic activities but low toxicity. Due to the high commercial value of the milbemycins and increasing resistance to the avermectins and their derivatives, it is imperative to develop an efficient combinatorial biosynthesis system exploiting an overproduction host strain to produce the milbemycins and novel analogs in large quantities...
January 17, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28095853/crispri-mediated-metabolic-engineering-of-e-coli-for-o-methylated-anthocyanin-production
#5
Brady F Cress, Quentin D Leitz, Daniel C Kim, Teresita D Amore, Jon Y Suzuki, Robert J Linhardt, Mattheos A G Koffas
BACKGROUND: Anthocyanins are a class of brightly colored, glycosylated flavonoid pigments that imbue their flower and fruit host tissues with hues of predominantly red, orange, purple, and blue. Although all anthocyanins exhibit pH-responsive photochemical changes, distinct structural decorations on the core anthocyanin skeleton also cause dramatic color shifts, in addition to improved stabilities and unique pharmacological properties. In this work, we report for the first time the extension of the reconstituted plant anthocyanin pathway from (+)-catechin to O-methylated anthocyanins in a microbial production system, an effort which requires simultaneous co-option of the endogenous metabolites UDP-glucose and S-adenosyl-L-methionine (SAM or AdoMet)...
January 17, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28093085/gene-design-fusion-technology-and-tev-cleavage-conditions-influence-the-purification-of-oxidized-disulphide-rich-venom-peptides-in-escherichia-coli
#6
Ana Filipa Sequeira, Jeremy Turchetto, Natalie J Saez, Fanny Peysson, Laurie Ramond, Yoan Duhoo, Marilyne Blémont, Vânia O Fernandes, Luís T Gama, Luís M A Ferreira, Catarina I P I Guerreiro, Nicolas Gilles, Hervé Darbon, Carlos M G A Fontes, Renaud Vincentelli
BACKGROUND: Animal venoms are large, complex libraries of bioactive, disulphide-rich peptides. These peptides, and their novel biological activities, are of increasing pharmacological and therapeutic importance. However, recombinant expression of venom peptides in Escherichia coli remains difficult due to the significant number of cysteine residues requiring effective post-translational processing. There is also an urgent need to develop high-throughput recombinant protocols applicable to the production of reticulated peptides to enable efficient screening of their drug potential...
January 17, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28068993/improvement-of-yeast-tolerance-to-acetic-acid-through-haa1-transcription-factor-engineering-towards-the-underlying-mechanisms
#7
Steve Swinnen, Sílvia F Henriques, Ranjan Shrestha, Ping-Wei Ho, Isabel Sá-Correia, Elke Nevoigt
BACKGROUND: Besides being a major regulator of the response to acetic acid in Saccharomyces cerevisiae, the transcription factor Haa1 is an important determinant of the tolerance to this acid. The engineering of Haa1 either by overexpression or mutagenesis has therefore been considered to be a promising avenue towards the construction of more robust strains with improved acetic acid tolerance. RESULTS: By applying the concept of global transcription machinery engineering to the regulon-specific transcription factor Haa1, a mutant allele containing two point mutations could be selected that resulted in a significantly higher acetic acid tolerance as compared to the wild-type allele...
January 9, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28068985/enzymatic-process-optimization-for-the-in-vitro-production-of-isoprene-from-mevalonate
#8
Tao Cheng, Hui Liu, Huibin Zou, Ningning Chen, Mengxun Shi, Congxia Xie, Guang Zhao, Mo Xian
BACKGROUND: As an important bulk chemical for synthetic rubber, isoprene can be biosynthesized by robust microbes. But rational engineering and optimization are often demanded to make the in vivo process feasible due to the complexities of cellular metabolism. Alternative synthetic biochemistry strategies are in fast development to produce isoprene or isoprenoids in vitro. RESULTS: This study set up an in vitro enzyme synthetic chemistry process using 5 enzymes in the lower mevalonate pathway to produce isoprene from mevalonate...
January 9, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28052753/chromosomal-position-effect-influences-the-heterologous-expression-of-genes-and-biosynthetic-gene-clusters-in-streptomyces-albus-j1074
#9
Bohdan Bilyk, Liliya Horbal, Andriy Luzhetskyy
BACKGROUND: Efforts to construct the Streptomyces host strain with enhanced yields of heterologous product have focussed mostly on engineering of primary metabolism and/or the deletion of endogenous biosynthetic gene clusters. However, other factors, such as chromosome compactization, have been shown to have a significant influence on gene expression levels in bacteria and fungi. The expression of genes and biosynthetic gene clusters may vary significantly depending on their location within the chromosome...
January 4, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28049530/metabolic-engineering-of-indole-pyruvic-acid-biosynthesis-in-escherichia-coli-with-tdid
#10
Yelin Zhu, Yan Hua, Biao Zhang, Lianhong Sun, Wenjie Li, Xin Kong, Jiong Hong
BACKGROUND: Indole pyruvic acid (IPA) is a versatile platform intermediate and building block for a number of high-value products in the pharmaceutical and food industries. It also has a wide range of applications, such as drugs for the nervous system, cosmetics, and luminophores. Chemical synthesis of IPA is a complicated and costly process. Moreover, through the biosynthesis route employing L-amino acid oxidase, the byproduct hydrogen peroxide leads the degradation of IPA. TdiD, identified as a specific tryptophan aminotransferase, could be an alternative solution for efficient IPA biosynthesis...
January 3, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28049528/improvement-of-whole-cell-transamination-with-saccharomyces-cerevisiae-using-metabolic-engineering-and-cell-pre-adaptation
#11
Nora Weber, Marie Gorwa-Grauslund, Magnus Carlquist
BACKGROUND: Whole-cell biocatalysis based on metabolically active baker's yeast with engineered transamination activity can be used to generate molecules carrying a chiral amine moiety. A prerequisite is though to express efficient ω-transaminases and to reach sufficient intracellular precursor levels. RESULTS: Herein, the efficiency of three different ω-transaminases originating from Capsicum chinense, Chromobacterium violaceum, and Ochrobactrum anthropi was compared for whole-cell catalyzed kinetic resolution of racemic 1-phenylethylamine to (R)-1-phenylethylamine...
January 3, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28049473/enhanced-heterologous-protein-productivity-by-genome-reduction-in-lactococcus-lactis-nz9000
#12
Duolong Zhu, Yuxin Fu, Fulu Liu, Haijin Xu, Per Erik Joakim Saris, Mingqiang Qiao
BACKGROUND: The implementation of novel chassis organisms to be used as microbial cell factories in industrial applications is an intensive research field. Lactococcus lactis, which is one of the most extensively studied model organisms, exhibits superior ability to be used as engineered host for fermentation of desirable products. However, few studies have reported about genome reduction of L. lactis as a clean background for functional genomic studies and a model chassis for desirable product fermentation...
January 3, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28031033/the-aceii-recombinant-trichoderma-reesei-qm9414-strains-with-enhanced-xylanase-production-and-its-applications-in-production-of-xylitol-from-tree-barks
#13
Lili Xiong, Ayyappa Kumar Sista Kameshwar, Xi Chen, Zhiyun Guo, Canquan Mao, Sanfeng Chen, Wensheng Qin
BACKGROUND: ACEII transcription factor plays a significant role in regulating the expression of cellulase and hemicellulase encoding genes. Apart from ACEII, transcription factors such as XYR1, CRE1, HAP2/3/5 complex and ACEI function in a coordinated pattern for regulating the gene expression of cellulases and hemicellulases. Studies have demonstrated that ACEII gene deletion results in decreased total cellulase and xylanase activities with reduced transcript levels of lignocellulolytic enzymes...
December 28, 2016: Microbial Cell Factories
https://www.readbyqxmd.com/read/28010736/isoprene-production-by-escherichia-coli-through-the-exogenous-mevalonate-pathway-with-reduced-formation-of-fermentation-byproducts
#14
Jung-Hun Kim, Chonglong Wang, Hui-Jung Jang, Myeong-Seok Cha, Ju-Eon Park, Seon-Yeong Jo, Eui-Sung Choi, Seon-Won Kim
BACKGROUND: Isoprene, a volatile C5 hydrocarbon, is an important platform chemical used in the manufacturing of synthetic rubber for tires and various other applications, such as elastomers and adhesives. RESULTS: In this study, Escherichia coli MG1655 harboring Populus trichocarpa isoprene synthase (PtispS) and the exogenous mevalonate (MVA) pathway produced 80 mg/L isoprene. Codon optimization and optimal expression of the ispS gene via adjustment of the RBS strength and inducer concentration increased isoprene production to 199 and 337 mg/L, respectively...
December 23, 2016: Microbial Cell Factories
https://www.readbyqxmd.com/read/28010734/proteins-adopt-functionally-active-conformations-after-type-iii-secretion
#15
Kevin James Metcalf, James Lea Bevington, Sandy Lisette Rosales, Lisa Ann Burdette, Elias Valdivia, Danielle Tullman-Ercek
BACKGROUND: Bacterial production of natively folded heterologous proteins by secretion to the extracellular space can improve protein production by simplifying purification and enabling continuous processing. In a typical bacterial protein production process, the protein of interest accumulates in the cytoplasm of the cell, requiring cellular lysis and extensive purification to separate the desired protein from other cellular constituents. The type III secretion system of Gram-negative bacteria is used to secrete proteins from the cytosol to the extracellular space in one step, but proteins must unfold during translocation, necessitating the folding of secreted proteins in the extracellular space for an efficient production process...
December 23, 2016: Microbial Cell Factories
https://www.readbyqxmd.com/read/27998268/mapping-the-membrane-proteome-of-anaerobic-gut-fungi-identifies-a-wealth-of-carbohydrate-binding-proteins-and-transporters
#16
Susanna Seppälä, Kevin V Solomon, Sean P Gilmore, John K Henske, Michelle A O'Malley
BACKGROUND: Engineered cell factories that convert biomass into value-added compounds are emerging as a timely alternative to petroleum-based industries. Although often overlooked, integral membrane proteins such as solute transporters are pivotal for engineering efficient microbial chassis. Anaerobic gut fungi, adapted to degrade raw plant biomass in the intestines of herbivores, are a potential source of valuable transporters for biotechnology, yet very little is known about the membrane constituents of these non-conventional organisms...
December 20, 2016: Microbial Cell Factories
https://www.readbyqxmd.com/read/27993152/copy-number-variability-of-expression-plasmids-determined-by-cell-sorting-and-droplet-digital-pcr
#17
Michael Jahn, Carsten Vorpahl, Thomas Hübschmann, Hauke Harms, Susann Müller
BACKGROUND: Plasmids are widely used for molecular cloning or production of proteins in laboratory and industrial settings. Constant modification has brought forth countless plasmid vectors whose characteristics in terms of average plasmid copy number (PCN) and stability are rarely known. The crucial factor determining the PCN is the replication system; most replication systems in use today belong to a small number of different classes and are available through repositories like the Standard European Vector Architecture (SEVA)...
December 19, 2016: Microbial Cell Factories
https://www.readbyqxmd.com/read/27978836/rhodococcus-aetherivorans-bcp1-as-cell-factory-for-the-production-of-intracellular-tellurium-nanorods-under-aerobic-conditions
#18
Alessandro Presentato, Elena Piacenza, Max Anikovskiy, Martina Cappelletti, Davide Zannoni, Raymond J Turner
BACKGROUND: Tellurite (TeO3(2-)) is recognized as a toxic oxyanion to living organisms. However, mainly anaerobic or facultative-anaerobic microorganisms are able to tolerate and convert TeO3(2-) into the less toxic and available form of elemental Tellurium (Te(0)), producing Te-deposits or Te-nanostructures. The use of TeO3(2-)-reducing bacteria can lead to the decontamination of polluted environments and the development of "green-synthesis" methods for the production of nanomaterials...
December 15, 2016: Microbial Cell Factories
https://www.readbyqxmd.com/read/27955649/engineering-aspergillus-niger-for-galactaric-acid-production-elimination-of-galactaric-acid-catabolism-by-using-rna-sequencing-and-crispr-cas9
#19
Joosu Kuivanen, Y-M Jasmin Wang, Peter Richard
BACKGROUND: meso-Galactaric acid is a dicarboxylic acid that can be produced by the oxidation of D-galacturonic acid, the main constituent of pectin. Mould strains can be engineered to perform this oxidation by expressing the bacterial enzyme uronate dehydrogenase. In addition, the endogenous pathway for D-galacturonic acid catabolism has to be inactivated. The filamentous fungus Aspergillus niger would be a suitable strain for galactaric acid production since it is efficient in pectin hydrolysis, however, it is catabolizing the resulting galactaric acid via an unknown catabolic pathway...
December 12, 2016: Microbial Cell Factories
https://www.readbyqxmd.com/read/27927208/use-of-a-sec-signal-peptide-library-from-bacillus-subtilis-for-the-optimization-of-cutinase-secretion-in-corynebacterium-glutamicum
#20
Johannes Hemmerich, Peter Rohe, Britta Kleine, Sarah Jurischka, Wolfgang Wiechert, Roland Freudl, Marco Oldiges
BACKGROUND: Technical bulk enzymes represent a huge market, and the extracellular production of such enzymes is favorable due to lowered cost for product recovery. Protein secretion can be achieved via general secretion (Sec) pathway. Specific sequences, signal peptides (SPs), are necessary to direct the target protein into the translocation machinery. For example, >150 Sec-specific SPs have been identified for Bacillus subtilis alone. As the best SP for a target protein of choice cannot be predicted a priori, screening of homologous SPs has been shown to be a powerful tool for different expression organisms...
December 7, 2016: Microbial Cell Factories
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