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Microbial Cell Factories

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https://www.readbyqxmd.com/read/28923077/development-and-evaluation-of-an-efficient-heterologous-gene-knock-in-reporter-system-in-lactococcus-lactis
#1
Yifei Lu, Hongxiang Yan, Jiezhong Deng, Zhigang Huang, Xurui Jin, Yanlan Yu, Qiwen Hu, Fuquan Hu, Jing Wang
BACKGROUND: Lactococcus lactis is a food grade probiotics and widely used to express heterologous proteins. Generally, target genes are knocked into the L. lactis genome through double-crossover recombination to express heterologous proteins stably. However, creating marker-less heterologous genes knocked-in clones is laborious. In this study, an efficient heterologous gene knock-in reporter system was developed in L. lactis NZ9000. RESULTS: Our knock-in reporter system consists of a temperature-sensitive plasmid pJW and a recombinant L...
September 18, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28903770/a-novel-bi-directional-promoter-system-allows-tunable-recombinant-protein-production-in-pichia-pastoris
#2
Vignesh Rajamanickam, Karl Metzger, Christian Schmid, Oliver Spadiut
BACKGROUND: The methylotrophic yeast Pichia pastoris is a well-studied host organism for recombinant protein production, which is usually regulated either by a constitutive promoter (e.g. promoter of glyceraldehyde-3-phosphate dehydrogenase; PGAP) or an inducible promoter (e.g. promoter of alcohol oxidase 1; PAOX1). Both promoter systems have several advantages and disadvantages; with one of the main disadvantages being their lack of tunability. Various novel promoter systems, which are either inducible or de-repressed, allowing higher degrees of freedom, have been reported...
September 13, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28903764/comparative-assessment-of-fermentative-capacity-of-different-xylose-consuming-yeasts
#3
Henrique César Teixeira Veras, Nádia Skorupa Parachin, João Ricardo Moreira Almeida
BACKGROUND: Understanding the effects of oxygen levels on yeast xylose metabolism would benefit ethanol production. In this work, xylose fermentative capacity of Scheffersomyces stipitis, Spathaspora passalidarum, Spathaspora arborariae and Candida tenuis was systematically compared under aerobic, oxygen-limited and anaerobic conditions. RESULTS: Fermentative performances of the four yeasts were greatly influenced by oxygen availability. S. stipitis and S. passalidarum showed the highest ethanol yields (above 0...
September 13, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28899391/mixomics-analysis-of-bacillus-subtilis-effect-of-oxygen-availability-on-riboflavin-production
#4
Junlang Hu, Pan Lei, Ali Mohsin, Xiaoyun Liu, Mingzhi Huang, Liang Li, Jianhua Hu, Haifeng Hang, Yingping Zhuang, Meijin Guo
BACKGROUND: Riboflavin, an intermediate of primary metabolism, is one kind of important food additive with high economic value. The microbial cell factory Bacillus subtilis has already been proven to possess significant importance for the food industry and have become one of the most widely used riboflavin-producing strains. In the practical fermentation processes, a sharp decrease in riboflavin production is encountered along with a decrease in the dissolved oxygen (DO) tension. Influence of this oxygen availability on riboflavin biosynthesis through carbon central metabolic pathways in B...
September 12, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28899372/the-combination-of-recombinant-and-non-recombinant-bacillus-subtilis-spore-display-technology-for-presentation-of-antigen-and-adjuvant-on-single-spore
#5
Wojciech Potocki, Alessandro Negri, Grażyna Peszyńska-Sularz, Krzysztof Hinc, Michał Obuchowski, Adam Iwanicki
BACKGROUND: Bacillus subtilis spores can be used for presentation of heterologous proteins. Two main approaches have been developed, the recombinant one, requiring modification of bacterial genome to express a protein of interest as a fusion with spore-coat protein, and non-recombinant, based on the adsorption of a heterologous protein onto the spore. So far only single proteins have been displayed on the spore surface. RESULTS: We have used a combined approach to adsorb and display FliD protein of Clostridium difficile on the surface of recombinant IL-2-presenting spores...
September 12, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28893251/identification-and-immobilization-of-a-novel-cold-adapted-esterase-and-its-potential-for-bioremediation-of-pyrethroid-contaminated-vegetables
#6
Xinjiong Fan, Weiqu Liang, Yanfang Li, He Li, Xiaolong Liu
BACKGROUND: Pyrethroids are potentially harmful to living organisms and ecosystems. Thus, concerns have been raised about pyrethroid residues and their persistence in agricultural products. To date, although several pyrethroid-hydrolyzing enzymes have been cloned, very few reports are available on pyrethroid-hydrolyzing enzymes with cold adaptation, high hydrolytic activity and good reusability, indispensable properties in practical bioremediation of pyrethroid-contaminated vegetables...
September 11, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28830549/lactobacillus-delivery-of-bioactive-interleukin-22
#7
Yin Lin, Kasper Krogh-Andersen, Lennart Hammarström, Harold Marcotte
BACKGROUND: Interleukin-22 (IL-22) plays a prominent role in epithelial regeneration and dampening of chronic inflammatory responses by protecting intestinal stem cells from immune-mediated tissue damage. IL-22 has a considerable therapeutic potential in graft-versus-host disease (GVHD), which is a frequent and challenging complication following allogeneic stem cell transplantation. The aim of our study was to engineer Lactobacillus for delivery of IL-22 directly to the intestinal mucosa as a new therapeutic strategy for GVHD...
August 23, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28830431/erratum-to-enhanced-biosurfactant-production-through-cloning-of-three-genes-and-role-of-esterase-in-biosurfactant-release
#8
Kamaljeet Kaur Sekhon, Sunil Khanna, Swaranjit Singh Cameotra
No abstract text is available yet for this article.
August 22, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28821247/a-cost-effective-approach-to-produce-15-n-labelled-amino-acids-employing-chlamydomonas-reinhardtii-cc503
#9
Jesús Nicolás Carcelén, Juan Manuel Marchante-Gayón, Pablo Rodríguez González, Luis Valledor, María Jesús Cañal, José Ignacio García Alonso
BACKGROUND: The use of enriched stable isotopes is of outstanding importance in chemical metrology as it allows the application of isotope dilution mass spectrometry (IDMS). Primary methods based on IDMS ensure the quality of the analytical measurements and traceability of the results to the international system of units. However, the synthesis of isotopically labelled molecules from enriched stable isotopes is an expensive and a difficult task. Either chemical and biochemical methods to produce labelled molecules have been proposed, but so far, few cost-effective methods have been described...
August 18, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28818103/enhancement-of-poly-3-hydroxybutyrate-co-3-hydroxyvalerate-accumulation-in-arxula-adeninivorans-by-stabilization-of-production
#10
Mateusz Biernacki, Marek Marzec, Thomas Roick, Reinhard Pätz, Kim Baronian, Rüdiger Bode, Gotthard Kunze
BACKGROUND: In recent years the production of biobased biodegradable plastics has been of interest of researchers partly due to the accumulation of non-biodegradable plastics in the environment and to the opportunity for new applications. Commonly investigated are the polyhydroxyalkanoates (PHAs) poly(hydroxybutyrate) and poly(hydroxybutyrate-co-hydroxyvalerate) (PHB-V). The latter has the advantage of being tougher and less brittle. The production of these polymers in bacteria is well established but production in yeast may have some advantages, e...
August 17, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28818083/optimization-of-the-biotechnological-production-of-a-novel-class-of-anti-mrsa-antibiotics-from-chitinophaga-sancti
#11
Amelie Beckmann, Stephan Hüttel, Viktoria Schmitt, Rolf Müller, Marc Stadler
BACKGROUND: Recently, the discovery of the elansolids, a group of macrolides, was reported. The molecules show activity against methicillin-resistant Staphylococcus aureus as well as other gram-positive organisms. This fact renders those substances a promising starting point for future chemical development. The active atropisomers A1/A2 are formed by macrolactonization of the biosynthesis product A3 but are prone to ring opening and subsequent formation of several unwanted side products...
August 17, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28818078/development-of-a-silicon-limitation-inducible-expression-system-for-recombinant-protein-production-in-the-centric-diatoms-thalassiosira-pseudonana-and-cyclotella-cryptica
#12
Roshan P Shrestha, Mark Hildebrand
BACKGROUND: An inducible promoter for recombinant protein expression provides substantial benefits because under induction conditions cellular energy and metabolic capability can be directed into protein synthesis. The most widely used inducible promoter for diatoms is for nitrate reductase, however, nitrogen metabolism is tied into diverse aspects of cellular function, and the induction response is not necessarily robust. Silicon limitation offers a means to eliminate energy and metabolic flux into cell division processes, with little other detrimental effect on cellular function, and a protein expression system that works under those conditions could be advantageous...
August 17, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28814300/increasing-on-target-cleavage-efficiency-for-crispr-cas9-induced-large-fragment-deletion-in-myxococcus-xanthus
#13
Ying-Jie Yang, Ye Wang, Zhi-Feng Li, Ya Gong, Peng Zhang, Wen-Chao Hu, Duo-Hong Sheng, Yue-Zhong Li
BACKGROUND: The CRISPR/Cas9 system is a powerful tool for genome editing, in which the sgRNA binds and guides the Cas9 protein for the sequence-specific cleavage. The protocol is employable in different organisms, but is often limited by cell damage due to the endonuclease activity of the introduced Cas9 and the potential off-target DNA cleavage from incorrect guide by the 20 nt spacer. RESULTS: In this study, after resolving some critical limits, we have established an efficient CRISPR/Cas9 system for the deletion of large genome fragments related to the biosynthesis of secondary metabolites in Myxococcus xanthus cells...
August 16, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28810867/new-inducible-promoter-for-gene-expression-and-synthetic-biology-in-yarrowia-lipolytica
#14
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
#15
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
#16
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
#17
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
#18
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
#19
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
#20
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
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