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Microbial Cell Factories

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https://www.readbyqxmd.com/read/28320377/application-of-an-e-coli-signal-sequence-as-a-versatile-inclusion-body-tag
#1
Wouter S P Jong, David Vikström, Diane Houben, H Bart van den Berg van Saparoea, Jan-Willem de Gier, Joen Luirink
BACKGROUND: Heterologous protein production in Escherichia coli often suffers from bottlenecks such as proteolytic degradation, complex purification procedures and toxicity towards the expression host. Production of proteins in an insoluble form in inclusion bodies (IBs) can alleviate these problems. Unfortunately, the propensity of heterologous proteins to form IBs is variable and difficult to predict. Hence, fusing the target protein to an aggregation prone polypeptide or IB-tag is a useful strategy to produce difficult-to-express proteins in an insoluble form...
March 21, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28302114/implications-of-evolutionary-engineering-for-growth-and-recombinant-protein-production-in-methanol-based-growth-media-in-the-yeast-pichia-pastoris
#2
Josef W Moser, Roland Prielhofer, Samuel M Gerner, Alexandra B Graf, Iain B H Wilson, Diethard Mattanovich, Martin Dragosits
BACKGROUND: Pichia pastoris is a widely used eukaryotic expression host for recombinant protein production. Adaptive laboratory evolution (ALE) has been applied in a wide range of studies in order to improve strains for biotechnological purposes. In this context, the impact of long-term carbon source adaptation in P. pastoris has not been addressed so far. Thus, we performed a pilot experiment in order to analyze the applicability and potential benefits of ALE towards improved growth and recombinant protein production in P...
March 17, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28302121/type-iis-restriction-based-combinatory-modulation-technique-for-metabolic-pathway-optimization
#3
Lijun Ye, Ping He, Qingyan Li, Xueli Zhang, Changhao Bi
BACKGROUND: One of the most important research subjects of metabolic engineering is pursuing a balanced metabolic pathway, which is the basis of an efficient cell factory. In this work, we dedicated to develop a simple and efficient technique to modulate expression of multiple genes simultaneously, and select for the optimal regulation pattern. RESULTS: A Type IIs restriction based combinatory modulation (TRCM) technique was designed and established in the research...
March 16, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28298234/dynamic-regulation-of-fatty-acid-pools-for-improved-production-of-fatty-alcohols-in-saccharomyces-cerevisiae
#4
Paulo Gonçalves Teixeira, Raphael Ferreira, Yongjin J Zhou, Verena Siewers, Jens Nielsen
BACKGROUND: In vivo production of fatty acid-derived chemicals in Saccharomyces cerevisiae requires strategies to increase the intracellular supply of either acyl-CoA or free fatty acids (FFAs), since their cytosolic concentrations are quite low in a natural state for this organism. Deletion of the fatty acyl-CoA synthetase genes FAA1 and FAA4 is an effective and straightforward way to disable re-activation of fatty acids and drastically increase FFA levels. However, this strategy causes FFA over-accumulation and consequential release to the extracellular medium, which results in a significant loss of precursors that compromises the process yield...
March 15, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28298224/transcriptional-reprogramming-in-yeast-using-dcas9-and-combinatorial-grna-strategies
#5
Emil D Jensen, Raphael Ferreira, Tadas Jakočiūnas, Dushica Arsovska, Jie Zhang, Ling Ding, Justin D Smith, Florian David, Jens Nielsen, Michael K Jensen, Jay D Keasling
BACKGROUND: Transcriptional reprogramming is a fundamental process of living cells in order to adapt to environmental and endogenous cues. In order to allow flexible and timely control over gene expression without the interference of native gene expression machinery, a large number of studies have focused on developing synthetic biology tools for orthogonal control of transcription. Most recently, the nuclease-deficient Cas9 (dCas9) has emerged as a flexible tool for controlling activation and repression of target genes, by the simple RNA-guided positioning of dCas9 in the vicinity of the target gene transcription start site...
March 15, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28298220/enzymatic-improvement-of-mitochondrial-thiol-oxidase-erv1-for-oxidized-glutathione-fermentation-by-saccharomyces-cerevisiae
#6
Jyumpei Kobayashi, Daisuke Sasaki, Kiyotaka Y Hara, Tomohisa Hasunuma, Akihiko Kondo
BACKGROUND: Oxidized glutathione (GSSG) is the preferred form for industrial mass production of glutathione due to its high stability compared with reduced glutathione (GSH). In our previous study, over-expression of the mitochondrial thiol oxidase ERV1 gene was the most effective for high GSSG production in Saccharomyces cerevisiae cells among three types of different thiol oxidase genes. RESULTS: We improved Erv1 enzyme activity for oxidation of GSH and revealed that S32 and N34 residues are critical for the oxidation...
March 15, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28288640/overexpression-of-a-c4-dicarboxylate-transporter-is-the-key-for-rerouting-citric-acid-to-c4-dicarboxylic-acid-production-in-aspergillus-carbonarius
#7
Lei Yang, Eleni Christakou, Jesper Vang, Mette Lübeck, Peter Stephensen Lübeck
BACKGROUND: C4-dicarboxylic acids, including malic acid, fumaric acid and succinic acid, are valuable organic acids that can be produced and secreted by a number of microorganisms. Previous studies on organic acid production by Aspergillus carbonarius, which is capable of producing high amounts of citric acid from varieties carbon sources, have revealed its potential as a fungal cell factory. Earlier attempts to reroute citric acid production into C4-dicarboxylic acids have been with limited success...
March 14, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28288625/functional-differentiation-of-3-ketosteroid-%C3%AE-1-dehydrogenase-isozymes-in-rhodococcus-ruber-strain-chol-4
#8
Govinda Guevara, Laura Fernández de Las Heras, Julián Perera, Juana María Navarro Llorens
BACKGROUND: The Rhodococcus ruber strain Chol-4 genome contains at least three putative 3-ketosteroid Δ(1)-dehydrogenase ORFs (kstD1, kstD2 and kstD3) that code for flavoenzymes involved in the steroid ring degradation. The aim of this work is the functional characterization of these enzymes prior to the developing of different biotechnological applications. RESULTS: The three R. ruber KstD enzymes have different substrate profiles. KstD1 shows preference for 9OHAD and testosterone, followed by progesterone, deoxy corticosterone AD and, finally, 4-BNC, corticosterone and 19OHAD...
March 14, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28279185/constitutively-stressed-yeast-strains-are-high-yielding-for-recombinant-fps1-implications-for-the-translational-regulation-of-an-aquaporin
#9
Stephanie P Cartwright, Richard A J Darby, Debasmita Sarkar, Nicklas Bonander, Stephane R Gross, Mark P Ashe, Roslyn M Bill
BACKGROUND: We previously selected four strains of Saccharomyces cerevisiae for their ability to produce the aquaporin Fps1 in sufficient yield for further study. Yields from the yeast strains spt3Δ, srb5Δ, gcn5Δ and yTHCBMS1 (supplemented with 0.5 μg/mL doxycycline) that had been transformed with an expression plasmid containing 249 base pairs of 5' untranslated region (UTR) in addition to the primary FPS1 open reading frame (ORF) were 10-80 times higher than yields from wild-type cells expressing the same plasmid...
March 9, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28259156/trends-in-recombinant-protein-use-in-animal-production
#10
REVIEW
Laia Gifre, Anna Arís, Àlex Bach, Elena Garcia-Fruitós
Recombinant technologies have made possible the production of a broad catalogue of proteins of interest, including those used for animal production. The most widely studied proteins for the animal sector are those with an important role in reproduction, feed efficiency, and health. Nowadays, mammalian cells and fungi are the preferred choice for recombinant production of hormones for reproductive purposes and fibrolytic enzymes to enhance animal performance, respectively. However, the development of low-cost products is a priority, particularly in livestock...
March 4, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28253886/cellular-factories-for-coenzyme-q10-production
#11
REVIEW
Sean Qiu En Lee, Tsu Soo Tan, Makoto Kawamukai, Ee Sin Chen
Coenzyme Q10 (CoQ10), a benzoquinone present in most organisms, plays an important role in the electron-transport chain, and its deficiency is associated with various neuropathies and muscular disorders. CoQ10 is the only lipid-soluble antioxidant found in humans, and for this, it is gaining popularity in the cosmetic and healthcare industries. To meet the growing demand for CoQ10, there has been considerable interest in ways to enhance its production, the most effective of which remains microbial fermentation...
March 2, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28245829/improving-escherichia-coli-membrane-integrity-and-fatty-acid-production-by-expression-tuning-of-fadl-and-ompf
#12
Zaigao Tan, William Black, Jong Moon Yoon, Jacqueline V Shanks, Laura R Jarboe
BACKGROUND: Construction of microbial biocatalysts for the production of biorenewables at economically viable yields and titers is frequently hampered by product toxicity. Membrane damage is often deemed as the principal mechanism of this toxicity, particularly in regards to decreased membrane integrity. Previous studies have attempted to engineer the membrane with the goal of increasing membrane integrity. However, most of these works focused on engineering of phospholipids and efforts to identify membrane proteins that can be targeted to improve fatty acid production have been unsuccessful...
February 28, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28245828/transcriptional-activator-cat8-is-involved-in-regulation-of-xylose-alcoholic-fermentation-in-the-thermotolerant-yeast-ogataea-hansenula-polymorpha
#13
Justyna Ruchala, Olena O Kurylenko, Nitnipa Soontorngun, Kostyantyn V Dmytruk, Andriy A Sibirny
BACKGROUND: Efficient xylose alcoholic fermentation is one of the key to a successful lignocellulosic ethanol production. However, regulation of this process in the native xylose-fermenting yeasts is poorly understood. In this work, we paid attention to the transcriptional factor Cat8 and its possible role in xylose alcoholic fermentation in Ogataea (Hansenula) polymorpha. In Saccharomyces cerevisiae, organism, which does not metabolize xylose, gene CAT8 encodes a Zn-cluster transcriptional activator necessary for expression of genes involved in gluconeogenesis, respiration, glyoxylic cycle and ethanol utilization...
February 28, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28245812/molecular-and-catalytic-properties-of-fungal-extracellular-cellobiose-dehydrogenase-produced-in-prokaryotic-and-eukaryotic-expression-systems
#14
Su Ma, Marita Preims, François Piumi, Lisa Kappel, Bernhard Seiboth, Eric Record, Daniel Kracher, Roland Ludwig
BACKGROUND: Cellobiose dehydrogenase (CDH) is an extracellular enzyme produced by lignocellulolytic fungi. cdh gene expression is high in cellulose containing media, but relatively low CDH concentrations are found in the supernatant of fungal cultures due to strong binding to cellulose. Therefore, heterologous expression of CDH in Pichia pastoris was employed in the last 15 years, but the obtained enzymes were over glycosylated and had a reduced specific activity. RESULTS: We compare the well-established CDH expression host P...
February 28, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28241831/nutrient-starvation-leading-to-triglyceride-accumulation-activates-the-entner-doudoroff-pathway-in-rhodococcus-jostii-rha1
#15
Antonio Juarez, Juan A Villa, Val F Lanza, Beatriz Lázaro, Fernando de la Cruz, Héctor M Alvarez, Gabriel Moncalián
BACKGROUND: Rhodococcus jostii RHA1 and other actinobacteria accumulate triglycerides (TAG) under nutrient starvation. This property has an important biotechnological potential in the production of sustainable oils. RESULTS: To gain insight into the metabolic pathways involved in TAG accumulation, we analysed the transcriptome of R jostii RHA1 under nutrient-limiting conditions. We correlate these physiological conditions with significant changes in cell physiology...
February 27, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28231787/ethylene-production-with-engineered-synechocystis-sp-pcc-6803-strains
#16
Vinod Puthan Veetil, S Andreas Angermayr, Klaas J Hellingwerf
BACKGROUND: Metabolic engineering and synthetic biology of cyanobacteria offer a promising sustainable alternative approach for fossil-based ethylene production, by using sunlight via oxygenic photosynthesis, to convert carbon dioxide directly into ethylene. Towards this, both well-studied cyanobacteria, i.e., Synechocystis sp PCC 6803 and Synechococcus elongatus PCC 7942, have been engineered to produce ethylene by introducing the ethylene-forming enzyme (Efe) from Pseudomonas syringae pv...
February 23, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28228147/artificially-designed-recombinant-protein-composed-of-multiple-epitopes-of-foot-and-mouth-disease-virus-as-a-vaccine-candidate
#17
Ho-Bin Lee, Da-Chuan Piao, Jun-Yeong Lee, Jae-Yun Choi, Jin-Duck Bok, Chong-Su Cho, Sang-Kee Kang, Yun-Jaie Choi
BACKGROUND: Concerns regarding the safety of inactivated foot-and-mouth disease (FMD) vaccine have been raised since it is produced from cultured live FMD virus (FMDV). To overcome this issue, recombinant protein has been studied as an alternative vaccine. RESULTS AND CONCLUSION: We designed a chimerical multi-epitope recombinant protein (5BT), which is comprised of tandem repeats of five B cell epitopes (residue of VP1 136-162) derived from different FMDV variants and one T-cell epitope (residue of 3A 21-35)...
February 22, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28219382/high-level-extracellular-protein-production-in-bacillus-subtilis-using-an-optimized-dual-promoter-expression-system
#18
Kang Zhang, Lingqia Su, Xuguo Duan, Lina Liu, Jing Wu
BACKGROUND: We recently constructed a Bacillus subtilis strain (CCTCC M 2016536) from which we had deleted the srfC, spoIIAC, nprE, aprE and amyE genes. This strain is capable of robust recombinant protein production and amenable to high-cell-density fermentation. Because the promoter is among the factors that influence the production of target proteins, optimization of the initial promoter, PamyQ from Bacillus amyloliquefaciens, should improve protein expression using this strain. This study was undertaken to develop a new, high-level expression system in B...
February 20, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28212656/using-a-vector-pool-containing-variable-strength-promoters-to-optimize-protein-production-in-yarrowia-lipolytica
#19
Rémi Dulermo, François Brunel, Thierry Dulermo, Rodrigo Ledesma-Amaro, Jérémy Vion, Marion Trassaert, Stéphane Thomas, Jean-Marc Nicaud, Christophe Leplat
BACKGROUND: The yeast Yarrowia lipolytica is an increasingly common biofactory. To enhance protein expression, several promoters have been developed, including the constitutive TEF promoter, the inducible POX2 promotor, and the hybrid hp4d promoter. Recently, new hp4d-inspired promoters have been created that couple various numbers of UAS1 tandem elements with the minimal LEU2 promoter or the TEF promoter. Three different protein-secretion signaling sequences can be used: preLip2, preXpr2, and preSuc2...
February 17, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28202018/de-novo-biosynthesis-of-pterostilbene-in-an-escherichia-coli-strain-using-a-new-resveratrol-o-methyltransferase-from-arabidopsis
#20
Kyung Taek Heo, Sun-Young Kang, Young-Soo Hong
BACKGROUND: Pterostilbene, a structural analog of resveratrol, has higher oral bioavailability and bioactivity than that of the parent compound; but is far less abundant in natural sources. Thus, to efficiently obtain this bioactive resveratrol analog, it is necessary to develop new bioproduction systems. RESULTS: We identified a resveratrol O-methyltransferase (ROMT) function from a multifunctional caffeic acid O-methyltransferase (COMT) originating from Arabidopsis, which catalyzes the transfer of a methyl group to resveratrol resulting in pterostilbene production...
February 15, 2017: Microbial Cell Factories
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