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Ciarán L Kelly, George M Taylor, Andrew Hitchcock, Antonio Torres-Méndez, John T Heap
Cyanobacteria are important for fundamental studies of photosynthesis and have great biotechnological potential. In order to better study and fully exploit these organisms, the limited repertoire of genetic tools and parts must be expanded. A small number of inducible promoters have been used in cyanobacteria, allowing dynamic external control of gene expression through the addition of specific inducer molecules. However, the inducible promoters used to date suffer from various drawbacks including toxicity of inducers, leaky expression in the absence of inducer and inducer photolability, the latter being particularly relevant to cyanobacteria which, as photoautotrophs, are grown under light...
March 15, 2018: ACS Synthetic Biology
Nicole Borth, Wei-Shou Hu
No abstract text is available yet for this article.
March 2018: Biotechnology Journal
Dagmara Jankowska, Tobias Heck, Mark Schubert, Alpaslan Yerlikaya, Christophe Weymuth, Daniel Rentsch, Irene Schober, Michael Richter
Lignin is the most abundant aromatic biopolymer functioning as an integral compound of woody materials. In its unmodified form it shows limited water solubility and is relatively unreactive. Thus, biotechnological lignin valorisation for high performance applications is highly underexploited. To expand its application, a new synthesis route towards novel concrete dispersing agents was developed. The route relies on a lignin functionalization by enzymatic transformation. Screening of lignin modifying systems resulted in functionalized lignin polymers with improved solubility in aqueous systems...
March 15, 2018: Chembiochem: a European Journal of Chemical Biology
Michael W Deem, Melia Elizabeth Bonomo
Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) constitute a multi-functional, constantly evolving immune system in bacteria and archaea cells. A heritable, molecular memory is generated of phage, plasmids, or other mobile genetic elements that attempt to attack the cell. This memory is used to recognize and interfere with subsequent invasions from the same genetic elements. This versatile prokaryotic tool has also been used to advance applications in biotechnology...
March 15, 2018: Physical Biology
Masatoshi Matsunami, Daiji Endo, Naruya Saitou, Hitoshi Suzuki, Manabu Onuma
The wood mouse (genus Apodemus ) is one of the most common rodents in broad-leaf forests in the temperate zone of the Palaearctic region. Molecular studies of wood mice have critically enhanced the understanding of their evolution and ancestral biogeographic events. However, their molecular data are currently only limited to partial mitochondrial sequences and a few genes. Therefore, we sequenced the wood mouse genome to facilitate the acquisition of useful resources for inferring their molecular evolution...
February 2018: Data in Brief
Christian E Ogaugwu, Qiuying Cheng, Annabeth Fieck, Ivy Hurwitz, Ravi Durvasula
Constitutively active promoter elements for heterologous protein production in Lactococcus lactis are scarce. Here, the promoter of the PTS-IIC gene cluster from L. lactis NZ3900 is described. This promoter was cloned upstream of an enhanced green fluorescent protein, GFPmut3a, and transformed into L. lactis. Transformants produced up to 13.5 μg of GFPmut3a per milliliter of log phase cells. Addition of cellobiose further increased the production of GFPmut3a by up to two-fold when compared to glucose. Analysis of mutations at two specific positions in the PTS-IIC promoter showed that a 'T' to 'G' mutation within the -35 element resulted in constitutive expression in glucose, while a 'C' at nucleotide 7 in the putative cre site enhanced promoter activity in cellobiose...
March 2018: Biotechnology Reports
Takeshi Takamatsu, Marouane Baslam, Takuya Inomata, Kazusato Oikawa, Kimiko Itoh, Takayuki Ohnishi, Tetsu Kinoshita, Toshiaki Mitsui
Chloroplasts, which perform photosynthesis, are one of the most important organelles in green plants and algae. Chloroplasts maintain an independent genome that includes important genes encoding their photosynthetic machinery and various housekeeping functions. Owing to its non-recombinant nature, low mutation rates, and uniparental inheritance, the chloroplast genome (plastome) can give insights into plant evolution and ecology and in the development of biotechnological and breeding applications. However, efficient methods to obtain high-quality chloroplast DNA (cpDNA) are currently not available, impeding powerful sequencing and further functional genomics research...
2018: Frontiers in Plant Science
Sheun Oshinbolu, Rachana Shah, Gary Finka, Mike Molloy, Mark Uden, Daniel G Bracewell
BACKGROUND: A current challenge in bioprocessing is the ability to analyse critical quality attributes such as aggregation without prior purification. This study evaluated the use of fluorescent dyes (Bis-ANS, SYPRO Orange, Thioflavin T and ProteoStat) to characterise mAb aggregates in Chinese hamster ovary clarified cultures. RESULTS: The null and mAb culture supernatants showed an increase in fluorescence intensity over the duration of the culture. The null cultures on day 14 saw a rapid increase in fluorescence intensity; day 10 to day 14, Bis-ANS and Thioflavin T had average increases of 21% and 48%, respectively, whereas ProteoStat and SYPRO Orange showed an average increase of 60%...
March 2018: Journal of Chemical Technology and Biotechnology
Atsumi Hirose, Takuya Kasai, Motohide Aoki, Tomonari Umemura, Kazuya Watanabe, Atsushi Kouzuma
Electrochemically active bacteria (EAB) receive considerable attention for their utility in bioelectrochemical processes. Although electrode potentials are known to affect the metabolic activity of EAB, it is unclear whether EAB are able to sense and respond to electrode potentials. Here, we show that, in the presence of a high-potential electrode, a model EAB Shewanella oneidensis MR-1 can utilize NADH-dependent catabolic pathways and a background formate-dependent pathway to achieve high growth yield. We also show that an Arc regulatory system is involved in sensing electrode potentials and regulating the expression of catabolic genes, including those for NADH dehydrogenase...
March 14, 2018: Nature Communications
Susan E Hepp, Gina M Borgo, Simina Ticau, Taro Ohkawa, Matthew D Welch
The baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV), the type species of alphabaculoviruses, is an enveloped DNA virus that infects lepidopteran insects and is commonly known as a vector for protein expression and cell transduction. AcMNPV belongs to a diverse group of viral and bacterial pathogens that target the host cell actin cytoskeleton during infection. AcMNPV is unusual, however, in that it absolutely requires actin translocation into the nucleus early in infection, and actin polymerization within the nucleus late in infection coincident with viral replication...
March 14, 2018: Journal of Virology
Yang Yuan, Shan-Shan Pan
BACKGROUND: Late exercise preconditioning (LEP) is confirmed to have a protective effect on acute cardiovascular stress. However, the mechanisms by which mitophagy participates in EP-induced cardioprotection remain unclear. LEP may involve mitophagy mediated by the receptors PARK2 gene-encoded E3 ubiquitin ligase (Parkin) and BCL2/adenovirus E1B 19 kDa protein-interacting protein 3 (Bnip3) to scavenge damaged mitochondria. METHODS: Our exercise preconditioning (EP) protocol involved four 10-minute periods of running, separated by 10-minute recovery intervals, plus a period of exhaustive running at 24 hours after EP...
March 9, 2018: Journal of Cardiovascular Pharmacology
Mengzhu Li, Hong Deng, Rui Ma, Huiying Luo, Bin Yao, Xiaoyun Su
Pyranose oxidase (POx) is a homotetrameric flavoprotein that catalyzes the oxidation of pyranose-configured sugars at position C-2 to corresponding 2-ketoaldoses. The wide substrate specificity makes POx potential for application in various biotechnological industries. In the present study we reported the gene cloning and heterologous expression of a POx from the basidiomycete Trametes sp. and functionally expressed the gene in Escherichia coli BL21(DE3). Based on sequence alignment, three residues were chosen for site-directed mutagenesis to obtain two single mutants (K312E and E539K) and two double mutants (T166A/E539K and K312E/E539K)...
March 13, 2018: AMB Express
Nestor D Ortega-de la Rosa, Jose L Vázquez-Vázquez, Sergio Huerta-Ochoa, Miquel Gimeno, Mariano Gutiérrez-Rojas
Acinetobacter species are identified as producing surface-active and emulsifying molecules known as bioemulsifiers. Production, characterization and stability of bioemulsifiers produced by Acinetobacter bouvetii UAM25 were studied. A. bouvetii UAM25 grew in three different carbon and energy sources: ethanol, a glycerol-hexadecane mixture and waste cooking oil in an airlift bioreactor, showing that bioemulsifier production was growth associated. The three purified bioemulsifiers were lipo-heteropolysaccharides of high molecular weight (4866 ± 533 and 462 ± 101 kDa)...
March 14, 2018: Bioprocess and Biosystems Engineering
Anja Kuenz, Susan Krull
Itaconic acid is one of the basic chemicals for the polymer industry, which can be produced on the basis of renewable raw materials. Since the middle of the twentieth century, itaconic acid has been produced industrially using the filamentous fungus Aspergillus terreus. But the demand for the organic acid is low due to the high production costs compared to alternative petrochemical manufactured raw materials. The high production costs are based on a low final titer, low productivities, and the usage of pure sugars, purified molasses, or starch hydrolysates, since the fungus reacts very sensitively to impurities in a culture medium...
March 13, 2018: Applied Microbiology and Biotechnology
Aleksandra Małachowska, Paweł Olszewski
BACKGROUND: Transcriptional control of gene expression is a widely utilized regulatory mechanism in synthetic biology, biotechnology and recombinant protein production. It is achieved by utilization of naturally occurring promoters responding to nutrients or chemicals. Despite their regulatory properties, these promoters often possess features which diminish their utility for biotechnology. High basal expression level and low induction ratio can be removed using genetic engineering techniques, although this process is often laborious and time-consuming...
March 13, 2018: Microbial Cell Factories
Jun Sun, Qingzhuo Wang, Yu Jiang, Zhiqiang Wen, Lirong Yang, Jianping Wu, Sheng Yang
BACKGROUND: The soil bacterium Pseudomonas putida KT2440 is a "generally recognized as safe"-certified strain with robust property and versatile metabolism. Thus, it is an ideal candidate for synthetic biology, biodegradation, and other biotechnology applications. The known genome editing approaches of Pseudomonas are suboptimal; thus, it is necessary to develop a high efficiency genome editing tool. RESULTS: In this study, we established a fast and convenient CRISPR-Cas9 method in P...
March 13, 2018: Microbial Cell Factories
Marco Alexander Fraatz, Michael Goldmann, Torsten Geissler, Egon Gross, Michael Backes, Jens-Michael Hilmer, Jakob Ley, Johanna Rost, Alexander Francke, Holger Zorn
A number of methyl-branched aldehydes impart interesting flavor impressions, and especially 12-methyltridecanal is a highly sought after flavoring compound for savory foods. Its smell is reminiscent of cooked meat and tallow. For the biotechnological production of 12-methyltridecanal, the literature was screened for fungi forming iso-fatty acids. Suitable organisms were identified and successfully grown in submerged cultures. The culture medium was optimized to increase the yields of branched fatty acids. A recombinant carboxylic acid reductase was used to reduce 12-methyltridecanoic acid to 12-methyltridecanal...
March 14, 2018: Journal of Agricultural and Food Chemistry
Carla Ribeiro, João Borges, Ana M S Costa, Vítor M Gaspar, Verónica de Zea Bermudez, João F Mano
Hollow multilayered capsules have shown massive potential for being used in the biomedical and biotechnology fields, in applications such as cellular internalization, intracellular trafficking, drug delivery, or tissue engineering. In particular, hollow microcapsules, developed by resorting to porous calcium carbonate sacrificial templates, natural-origin building blocks and the prominent Layer-by-Layer (LbL) technology, have attracted increasing attention owing to their key features. However, these microcapsules revealed a great tendency to aggregate, which represents a major hurdle when aiming for cellular internalization and intracellular therapeutics delivery...
March 10, 2018: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
Daniel González-Mancebo, Ana I Becerro, T Cristina Rojas, Andrés Olivencia, Ariadna Corral, Marcin Balcerzyk, Eugenio Cantelar, Fernando Cussó, Manuel Ocaña
The singular properties of lanthanide-based inorganic nanoparticles (NPs) has raised the attention of the scientific community in biotechnological applications. In particular, those systems with two or more functionalities are especially interesting. In this work, an effective and commercially attractive procedure has been developed that renders uniform, water-dispersible Ln3+ :CeF3 (Ln = Tb, Nd) NPs with different shapes and size. The method consists of the homogeneous precipitation, in a mixture of polyol and water, of cations and anions using precursors that allow the controlled release of the latter...
March 5, 2018: Journal of Colloid and Interface Science
Javier Viña-Gonzalez, Katarina Elbl, Xavier Ponte, Francisco Valero, Miguel Alcalde
Aryl-alcohol oxidase (AAO) plays a fundamental role in the fungal ligninolytic secretome, acting as a supplier of H2 O2 . Despite its highly selective mechanism of action, the presence of this flavooxidase in different biotechnological settings has hitherto been hampered by the lack of appropriate heterologous expression systems. We recently described the functional expression of the AAO from Pleurotus eryngii in Saccharomyces cerevisiae by fusing a chimeric signal peptide (preαproK) and applying structure-guided evolution...
March 13, 2018: Biotechnology and Bioengineering
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