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Microbial Cell Factories

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https://www.readbyqxmd.com/read/28810867/new-inducible-promoter-for-gene-expression-and-synthetic-biology-in-yarrowia-lipolytica
#1
Marion Trassaert, Marie Vandermies, Fréderic Carly, Olivia Denies, Stéphane Thomas, Patrick Fickers, Jean-Marc Nicaud
BACKGROUND: The oleaginous yeast Yarrowia lipolytica is increasingly used as alternative cell factory for the production of recombinant proteins. At present, several promoters with different strengths have been developed based either on the constitutive pTEF promoter or on oleic acid inducible promoters such as pPOX2 and pLIP2. Although these promoters are highly efficient, there is still a lack of versatile inducible promoters for gene expression in Y. lipolytica. RESULTS: We have isolated and characterized the promoter of the EYK1 gene coding for an erythrulose kinase...
August 15, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28806958/responses-of-synechocystis-sp-pcc-6803-to-heterologous-biosynthetic-pathways
#2
Konstantinos Vavitsas, Emil Østergaard Rue, Lára Kristín Stefánsdóttir, Thiyagarajan Gnanasekaran, Andreas Blennow, Christoph Crocoll, Steinn Gudmundsson, Poul Erik Jensen
BACKGROUND: There are an increasing number of studies regarding genetic manipulation of cyanobacteria to produce commercially interesting compounds. The majority of these works study the expression and optimization of a selected heterologous pathway, largely ignoring the wholeness and complexity of cellular metabolism. Regulation and response mechanisms are largely unknown, and even the metabolic pathways themselves are not fully elucidated. This poses a clear limitation in exploiting the rich biosynthetic potential of cyanobacteria...
August 15, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28793898/erratum-to-a-review-on-lactococcus-lactis-from-food-to-factory
#3
Adelene Ai-Lian Song, Lionel L A In, Swee Hua Erin Lim, Raha Abdul Rahim
No abstract text is available yet for this article.
August 9, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28784139/structure-and-application-of-antifreeze-proteins-from-antarctic-bacteria
#4
Patricio A Muñoz, Sebastián L Márquez, Fernando D González-Nilo, Valeria Márquez-Miranda, Jenny M Blamey
BACKGROUND: Antifreeze proteins (AFPs) production is a survival strategy of psychrophiles in ice. These proteins have potential in frozen food industry avoiding the damage in the structure of animal or vegetal foods. Moreover, there is not much information regarding the interaction of Antarctic bacterial AFPs with ice, and new determinations are needed to understand the behaviour of these proteins at the water/ice interface. RESULTS: Different Antarctic places were screened for antifreeze activity and microorganisms were selected for the presence of thermal hysteresis in their crude extracts...
August 7, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28778165/highly-effective-regiospecific-reduction-of-chalcone-by-cyanobacteria-leads-to-the-formation-of-dihydrochalcone-two-steps-towards-natural-sweetness
#5
Beata Żyszka, Mirosław Anioł, Jacek Lipok
BACKGROUND: Chalcones are the biogenetic precursors of all known flavonoids, which play an essential role in various metabolic processes in photosynthesizing organisms. The use of whole cyanobacteria cells in a two-step, light-catalysed regioselective bio-reduction of chalcone, leading to the formation of the corresponding dihydrochalcone, is reported. The prokaryotic microalgae cyanobacteria are known to produce phenolic compounds, including flavonoids, as natural components of cells...
August 4, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28764701/a-versatile-one-step-crispr-cas9-based-approach-to-plasmid-curing
#6
Ida Lauritsen, Andreas Porse, Morten O A Sommer, Morten H H Nørholm
BACKGROUND: Plasmids are widely used and essential tools in molecular biology. However, plasmids often impose a metabolic burden and are only temporarily useful for genetic engineering, bio-sensing and characterization purposes. While numerous techniques for genetic manipulation exist, a universal tool enabling rapid removal of plasmids from bacterial cells is lacking. RESULTS: Based on replicon abundance and sequence conservation analysis, we show that the vast majority of bacterial cloning and expression vectors share sequence similarities that allow for broad CRISPR-Cas9 targeting...
August 2, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28764719/a-combination-of-hplc-and-automated-data-analysis-for-monitoring-the-efficiency-of-high-pressure-homogenization
#7
Britta Eggenreich, Vignesh Rajamanickam, David Johannes Wurm, Jens Fricke, Christoph Herwig, Oliver Spadiut
BACKGROUND: Cell disruption is a key unit operation to make valuable, intracellular target products accessible for further downstream unit operations. Independent of the applied cell disruption method, each cell disruption process must be evaluated with respect to disruption efficiency and potential product loss. Current state-of-the-art methods, like measuring the total amount of released protein and plating-out assays, are usually time-delayed and involve manual intervention making them error-prone...
August 1, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28754115/in-vivo-plug-and-play-a-modular-multi-enzyme-single-cell-catalyst-for-the-asymmetric-amination-of-ketoacids-and-ketones
#8
Judith E Farnberger, Elisabeth Lorenz, Nina Richter, Volker F Wendisch, Wolfgang Kroutil
BACKGROUND: Transaminases have become a key tool in biocatalysis to introduce the amine functionality into a range of molecules like prochiral α-ketoacids and ketones. However, due to the necessity of shifting the equilibrium towards the product side (depending on the amine donor) an efficient amination system may require three enzymes. So far, this well-established transformation has mainly been performed in vitro by assembling all biocatalysts individually, which comes along with elaborate and costly preparation steps...
July 28, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28754100/identifying-genomic-targets-for-protein-over-expression-by-omics-analysis-of-quiescent-escherichia-coli-cultures
#9
Shubhashree Mahalik, Ashish Kumar Sharma, Priyanka Jain, Krishna Jyoti Mukherjee
BACKGROUND: A cellular stress response is triggered upon induction of recombinant protein expression which feedback inhibits both growth as well as protein synthesis. In order to separate these two effects, it was decided to study "quiescent cultures" which continue to be metabolically active and express recombinant proteins even after growth cessation. The idea was to identify and up-regulate genes which are responsible for protein synthesis in the absence of growth. This would ensure that, even if growth were adversely affected post induction, there would be no attendant reduction in the protein expression capability of the cells...
July 28, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28750668/from-hybridomas-to-a-robust-microalgal-based-production-platform-molecular-design-of-a-diatom-secreting-monoclonal-antibodies-directed-against-the-marburg-virus-nucleoprotein
#10
Franziska Hempel, Michael Maurer, Björn Brockmann, Christian Mayer, Nadine Biedenkopf, Anne Kelterbaum, Stephan Becker, Uwe G Maier
BACKGROUND: The ideal protein expression system should provide recombinant proteins in high quality and quantity involving low production costs only. However, especially for complex therapeutic proteins like monoclonal antibodies many challenges remain to meet this goal and up to now production of monoclonal antibodies is very costly and delicate. Particularly, emerging disease outbreaks like Ebola virus in Western Africa in 2014-2016 make it necessary to reevaluate existing production platforms and develop robust and cheap alternatives that are easy to handle...
July 27, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28743272/establishment-of-neurospora-crassa-as-a-host-for-heterologous-protein-production-using-a-human-antibody-fragment-as-a-model-product
#11
David Havlik, Ulrike Brandt, Kathrin Bohle, André Fleißner
BACKGROUND: Filamentous fungi are commonly used as production hosts for bulk enzymes in biotechnological applications. Their robust and quick growth combined with their ability to secrete large amounts of protein directly into the culture medium makes fungi appealing organisms for the generation of novel production systems. The red bread mold Neurospora crassa has long been established as a model system in basic research. It can be very easily genetically manipulated and a wealth of molecular tools and mutants are available...
July 25, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28743271/development-of-inducer-free-expression-plasmids-based-on-iptg-inducible-promoters-for-bacillus-subtilis
#12
Dinh Thi Minh Tran, Trang Thi Phuong Phan, Thanh Kieu Huynh, Ngan Thi Kim Dang, Phuong Thi Kim Huynh, Tri Minh Nguyen, Tuom Thi Tinh Truong, Thuoc Linh Tran, Wolfgang Schumann, Hoang Duc Nguyen
BACKGROUND: Besides Escherichia coli, Bacillus subtilis is an important bacterial species for the production of recombinant proteins. Recombinant genes are inserted into shuttle expression vectors which replicate in both E. coli and in B. subtilis. The ligation products are first transformed into E. coli cells, analyzed for correct insertions, and the correct recombinant plasmids are then transformed into B. subtilis. A major problem using E. coli cells can be the strong basal level of expression of the recombinant protein which may interfere with the stability of the cells...
July 25, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28743267/production-of-a-recombinant-phospholipase-a2-in-escherichia-coli-using-resonant-acoustic-mixing-that-improves-oxygen-transfer-in-shake-flasks
#13
Norma A Valdez-Cruz, Greta I Reynoso-Cereceda, Saumel Pérez-Rodriguez, Sara Restrepo-Pineda, Jesus González-Santana, Alejandro Olvera, Guadalupe Zavala, Alejandro Alagón, Mauricio A Trujillo-Roldán
BACKGROUND: Shake flasks are widely used during the development of bioprocesses for recombinant proteins. Cultures of recombinant Escherichia coli with orbital mixing (OM) have an oxygen limitation negatively affecting biomass growth and recombinant-protein production. With the aim to improve mixing and aeration in shake flask cultures, we analyzed cultures subjected to OM and the novel resonant acoustic mixing (RAM) by applying acoustic energy to E. coli BL21-Gold (DE3): a producer of recombinant phospholipase A2 (rPLA2) from Micrurus laticollaris snake venom...
July 25, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28738864/identification-of-two-organohalide-respiring-dehalococcoidia-associated-to-different-dechlorination-activities-in-pcb-impacted-marine-sediments
#14
Andrea Nuzzo, Andrea Negroni, Giulio Zanaroli, Fabio Fava
BACKGROUND: Microbial reductive dechlorination of polychlorinated biphenyls (PCBs) plays a major role in detoxifying anoxic contaminated freshwater and marine sediments from PCBs. Known members of the phylum Chloroflexi are typically responsible for this activity in freshwater sediments, whereas less is known about the microorganisms responsible for this activity in marine sediments. PCB-respiring activities were detected in PCB-impacted marine sediments of the Venice Lagoon. The aim of this work was to identify the indigenous organohalide-respiring microorganisms in such environments and assess their dechlorination specificity against spiked Aroclor™ 1254 PCBs under laboratory conditions resembling the in situ biogeochemistry...
July 24, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28738851/expression-and-secretion-of-fungal-endoglucanase-ii-and-chimeric-cellobiohydrolase-i-in-the-oleaginous-yeast-lipomyces-starkeyi
#15
Qi Xu, Eric P Knoshaug, Wei Wang, Markus Alahuhta, John O Baker, Shihui Yang, Todd Vander Wall, Stephen R Decker, Michael E Himmel, Min Zhang, Hui Wei
BACKGROUND: Lipomyces starkeyi is one of the leading lipid-producing microorganisms reported to date; its genetic transformation was only recently reported. Our aim is to engineer L. starkeyi to serve in consolidated bioprocessing (CBP) to produce lipid or fatty acid-related biofuels directly from abundant and low-cost lignocellulosic substrates. RESULTS: To evaluate L. starkeyi in this role, we first conducted a genome analysis, which revealed the absence of key endo- and exocellulases in this yeast, prompting us to select and screen four signal peptides for their suitability for the overexpression and secretion of cellulase genes...
July 24, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28724440/bottleneck-in-secretion-of-%C3%AE-amylase-in-bacillus-subtilis
#16
REVIEW
Shaomin Yan, Guang Wu
Amylase plays an important role in biotechnology industries, and Gram-positive bacterium Bacillus subtilis is a major host to produce heterogeneous α-amylases. However, the secretion stress limits the high yield of α-amylase in B. subtilis although huge efforts have been made to address this secretion bottleneck. In this question-oriented review, every effort is made to answer the following questions, which look simple but are long-standing, through reviewing of literature: (1) Does α-amylase need a specific and dedicated chaperone? (2) What signal sequence does CsaA recognize? (3) Does CsaA require ATP for its operation? (4) Does an unfolded α-amylase is less soluble than a folded one? (5) Does α-amylase aggregate before transporting through Sec secretion system? (6) Is α-amylase sufficient stable to prevent itself from misfolding? (7) Does α-amylase need more disulfide bonds to be stabilized? (8) Which secretion system does PrsA pass through? (9) Is PrsA ATP-dependent? (10) Is PrsA reused after folding of α-amylase? (11) What is the fate of PrsA? (12) Is trigger factor (TF) ATP-dependent? The literature review suggests that not only the most of those questions are still open to answers but also it is necessary to calculate ATP budget in order to better understand how B...
July 19, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28724386/engineering-microbial-cell-factories-for-the-production-of-plant-natural-products-from-design-principles-to-industrial-scale-production
#17
REVIEW
Xiaonan Liu, Wentao Ding, Huifeng Jiang
Plant natural products (PNPs) are widely used as pharmaceuticals, nutraceuticals, seasonings, pigments, etc., with a huge commercial value on the global market. However, most of these PNPs are still being extracted from plants. A resource-conserving and environment-friendly synthesis route for PNPs that utilizes microbial cell factories has attracted increasing attention since the 1940s. However, at the present only a handful of PNPs are being produced by microbial cell factories at an industrial scale, and there are still many challenges in their large-scale application...
July 19, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28720107/reversible-bacterial-immobilization-based-on-the-salt-dependent-adhesion-of-the-bacterionanofiber-protein-ataa
#18
Shogo Yoshimoto, Yuki Ohara, Hajime Nakatani, Katsutoshi Hori
BACKGROUND: Immobilization of microbial cells is an important strategy for the efficient use of whole-cell catalysts because it simplifies product separation, enables the cell concentration to be increased, stabilizes enzymatic activity, and permits repeated or continuous biocatalyst use. However, conventional immobilization methods have practical limitations, such as limited mass transfer in the inner part of a gel, gel fragility, cell leakage from the support matrix, and adverse effects on cell viability and catalytic activity...
July 18, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28716098/awakening-sleeping-beauty-production-of-propionic-acid-in-escherichia-coli-through-the-sbm-operon-requires-the-activity-of-a-methylmalonyl-coa-epimerase
#19
Ricardo Axayacatl Gonzalez-Garcia, Tim McCubbin, Annalena Wille, Manuel Plan, Lars Keld Nielsen, Esteban Marcellin
BACKGROUND: Propionic acid is used primarily as a food preservative with smaller applications as a chemical building block for the production of many products including fabrics, cosmetics, drugs, and plastics. Biological production using propionibacteria would be competitive against chemical production through hydrocarboxylation of ethylene if native producers could be engineered to reach near-theoretical yield and good productivity. Unfortunately, engineering propionibacteria has proven very challenging...
July 17, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28716035/systematic-evaluation-of-characteristics-of-the-membrane-based-fed-batch-shake-flask
#20
P Philip, K Meier, D Kern, J Goldmanns, F Stockmeier, C Bähr, J Büchs
BACKGROUND: The initial part of process development involves extensive screening programs to identify optimal biological systems and cultivation conditions. For a successful scale-up, the operation mode on screening and production scale must be as close as possible. To enable screening under fed-batch conditions, the membrane-based fed-batch shake flask was developed. It is a shake flask mounted with a central feed reservoir with an integrated rotating membrane tip for a controlled substrate release...
July 17, 2017: Microbial Cell Factories
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