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  • VOLATILE FATTY ACIDS FROM ORGANIC WASTES AS NOVEL LOW-COST CARBON SOURCE FOR yarrowia lipolytica
    New Biotechnol. (IF 3.739) Pub Date : 2020-01-14
    Mercedes Llamas; Elia Tomás-Pejó; Cristina González-Fernández

    Microbial oil biosynthesis is envisaged as a promising technology for sustainable production of chemicals and fuels. Sugar-based substrates are the most typical carbon sources used for this purpose where metabolic pathways and stoichiometry are well known. However, the use of low-cost substrates is crucial for the economic viability of the process. Volatile fatty acids (VFAs) are considered to be a novel low-cost carbon source for microbial lipid production. They can be utilized by oleaginous yeasts to produce and store fatty acids in form of intracellular lipid bodies. In this work, Yarrowia lipolytica growth and substrate consumption were evaluated using the major VFAs present in anaerobic effluents. Individual VFAs as well as synthetic mixtures were tested at different concentrations to determine uptake rates and potential toxicity. Increasing VFA chain length resulted in greater biomass yield although, when added individually, 4 g Carbon/L VFA (e.g. 6.45 g/L of caproic and 10 g/L of acetic acid) caused inhibitory effects. Remarkably, biomass growth increased by 2.5-fold on real anaerobic fermentation effluent compared with synthetic mixtures. When real digestate was supplemented with synthetic VFAs up to 26.5 g/L, the inhibitory effect of the acids was counterbalanced. The results provided evidence of robustness of Y. lipolytica towards low-cost fermentation effluents and present this yeast as a promising candidate for the sustainable production of microbial oil using real digestates.

    更新日期:2020-01-15
  • Characterization of heavy metal toxicity in some plants and microorganisms – a preliminary approach for environmental bioremediation
    New Biotechnol. (IF 3.739) Pub Date : 2020-01-13
    Mariana Diaconu; Lucian Vasile Pavel; Raluca-Maria Hlihor; Mihaela Rosca; Daniela Ionela Fertu; Markus Lenz; Philippe Xavier Corvini; Maria Gavrilescu
    更新日期:2020-01-13
  • Calcium effect on microbial activity and biomass aggregation during anaerobic digestion at high salinity
    New Biotechnol. (IF 3.739) Pub Date : 2020-01-07
    Maria Cristina Gagliano; Dainis Sudmalis; Hardy Temmink; Caroline M. Plugge
    更新日期:2020-01-07
  • Microbial inoculum development for ameliorating crop drought stress: a case study of Variovorax paradoxus 5C-2
    New Biotechnol. (IF 3.739) Pub Date : 2019-12-30
    Rosalia Garcia Teijeiro; Andrey A. Belimov; Ian C. Dodd

    Drought affects plant hormonal homeostasis, including root to shoot signalling. The plant is intimately connected below-ground with soil-dwelling microbes, including plant growth promoting rhizobacteria (PGPR) that can modulate plant hormonal homeostasis. Incorporating PGPR into the rhizosphere often delivers favourable results in greenhouse experiments, while field applications are much less predictable. We review the natural processes that affect the formation and dynamics of the rhizosphere, establishing a model for successful field application of PGPR utilizing an example microbial inoculum, Variovorax paradoxus 5C-2.

    更新日期:2019-12-30
  • Microbiome changes and oxidative capability of an anaerobic PCB dechlorinating enrichment culture after oxygen exposure
    New Biotechnol. (IF 3.739) Pub Date : 2019-12-24
    Bruna Matturro; Giuseppe Mascolo; Simona Rossetti

    Marine sediments may represent a sink of persistent organic pollutants including polychlorinated biphenyls (PCBs), toxic compounds prone to reductive or oxidative biodegradation pathways depending on the degree of chlorination and the positions of the chlorine atoms on the biphenyl rings. Superficial marine sediments can be subjected to episodic sediment resuspension by boat traffic and wind action causing the exposure of the underlying anaerobic layer to oxygen. Under these dynamic conditions, a deeper knowledge of the adaptation capability of the autochthonous microbial communities towards severe changes of the reaction environment is required. Insights into the metabolic potential of sediment community members may contribute greatly to the definition of efficient and reliable in situ bioremediation strategies. In this study, an anaerobic PCB-dechlorinating microbial consortium, developed from the chronically polluted marine sediment of Mar Piccolo (Taranto, Italy), was used to evaluate the response of the sediment microbiome to the imposition of aerobic conditions after prolonged anaerobic incubation. Compared to the anaerobic control, a dramatic change in microbiome composition, with a marked increase of Alphaproteobacteria of up to 39.2% of total operational taxonomic units (OTUs) was revealed by high-throughput 16S rRNA gene sequencing. Accordingly, a decrement of low chlorinated PCBs (up to 58.3 ± 7.5 % for PCB 18) and the concomitant appearance of genes coding for PCB-degrading biphenyl dioxygenase (bph) were observed at the end of the aerobic incubation, suggesting the occurrence of oxidative PCB biodegradation processes.

    更新日期:2019-12-25
  • The Self-Forming Dynamic Membrane BioReactor (SFD MBR) as a suitable technology for agro-industrial wastewater treatment
    New Biotechnol. (IF 3.739) Pub Date : 2019-12-23
    Pompilio Vergine; Carlo Salerno; Giovanni Berardi; Giuseppe Pappagallo; Alfieri Pollice

    Two bench-scale Self-Forming Dynamic Membrane BioReactors (SFD MBR), equipped with 50 µm nylon meshes were set up and operated under aerobic conditions in order to treat canning and winery wastewaters. The results showed different behaviors of the two systems, confirming the strong dependence of SFD MBR performance on the type of biomass and, in turn, on the type of stream being treated. Both plants achieved good results in terms of effluent quality, demonstrating the suitability of the proposed technology. Median values of effluent turbidity were 2.7 and 15.4 NTU (Nephelometric Turbidity Units) in the reactors fed with canning wastewater and winery wastewater, respectively. The removal of organic matter (as COD, Chemical Oxygen Demand) was consistently above 90%, although the retention of suspended solids was variable and somewhat dependent on operating conditions and feed composition. The activated sludge characteristics were observed to affect filtration performance and in particular the capillary suction time (CST) was a possible indicator of efficiency, with a threshold value of 11 seconds above which filtration performance decreased. This parameter is proposed as an early warning tool for changes in the filtration performance of an SFD MBR, both for effluent quality and cleaning requirements.

    更新日期:2019-12-23
  • Enrichment of a mixed microbial culture of PHA-storing microorganisms by using fermented hardwood spent sulfite liquor
    New Biotechnol. (IF 3.739) Pub Date : 2019-12-12
    Joana Pereira; Diogo Queirós; Paulo C. Lemos; Simona Rossetti; Luísa S. Serafim

    Pulp and paper factories produce several residues that can be explored and valorized through polyhydroxyalkanoate (PHA) production via a three-step process. The objective of this work was focused on the selection step. Acidified hardwood spent sulfite liquor (HSSL), a fermented waste stream from a pulp and paper factory, was used to select a mixed microbial culture (MMC) in a sequencing batch reactor (SBR) operated for 156 days under different operational conditions. The MMC adapted to the imposed conditions, revealing its robustness whenever the operational parameters were changed. Feast-to-Famine ratio was kept below or equal to 0.2, with constant production of a copolymer of P(3HB-co-3 HV), and with storage contents values over 30%. Changes in the operational conditions, namely cycle length, and organic load rate (OLR), successfully led to the selection of an MMC with a stable accumulation capacity and an increased biomass concentration. Next Generation Sequencing analysis was performed on samples collected during the SBR operational period. The analysis of the microbial composition of the MMC showed a rise in PHA-accumulating bacteria over time. Acidovorax and Comamonas species were found mainly to drive the PHA storage process during the first two periods of operation. After an increase in the OLR, in the last period, a shift towards Comamonas dominance occurred, suggesting a higher tolerance to the inhibitory compounds of the HSSL for this genus.

    更新日期:2019-12-13
  • Aerobic granular-based technology for water and energy recovery from municipal wastewater
    New Biotechnol. (IF 3.739) Pub Date : 2019-12-11
    Marco De Sanctis, Valerio Guido Altieri, Valentina Piergrossi, Claudio Di Iaconi

    In the present study, the possibility of recovering both thermal energy and water for agricultural purposes from sewage is evaluated. A treatment plant, based on a sequencing batch biofilter granular reactor (SBBGR) followed by sand filtration and coupled with a solar wastewater source heat pump, was operated from September to November 2018 at a set-point temperature of 14 °C to verify the stability of heat recovery efficiency and the suitability of plant effluent to be reused for irrigation. Heat recovery did not influence the SBBGR treatment and disinfection efficiency, which removed about 90% of suspended solids, chemical and biochemical oxygen demand and ammonia, as well as 70% of total nitrogen, 3 log10 units of Escherichia coli and more than 1 log10 unit of Clostridium perfringens. Furthermore, after sand filtration, water quality complied with the standards for agricultural reuse currently in force in several countries. Energy extracted from SBBGR was mainly influenced by environmental conditions, affecting wastewater temperature, and also by wastewater composition, affecting the energy release due to bacterial metabolic activity for carbon and nitrogen removal. Notably, no evident deterioration of energy extraction efficiency from the SBBGR was observed, suggesting negligible fouling phenomena on the submerged thermal exchanger.

    更新日期:2019-12-11
  • Making plants into cost-effective bioreactors for highly active antimicrobial peptides
    New Biotechnol. (IF 3.739) Pub Date : 2019-12-05
    Meron Ghidey, S M Ashiqul Islam, Grace Pruett, Christopher Michel Kearney

    As antibiotic-resistant bacterial pathogens become an ever-increasing concern, antimicrobial peptides (AMPs) have grown increasingly attractive as alternatives. Potentially, plants could be used as cost-effective AMP bioreactors; however, reported heterologous AMP expression is much lower in plants than in E. coli expression systems and often results in plant cytotoxicity, even for AMPs fused to carrier proteins. This suggests that there may be a physical characteristic of the previously described heterologous AMPs which impedes efficient expression in plants. Using a meta-analysis of protein databases, this study has determined that native plant AMPs were significantly less cationic than AMPs native to other taxa. To apply this finding to plant expression, the transient expression of 10 different heterologous AMPs, ranging in charge from +7 to -5, was tested in the tobacco, Nicotiana benthamiana. Elastin-like polypeptide (ELP) was used as the carrier protein for AMP expression. ELP fusion allowed for a simple, cost-effective temperature shift purification. Using this system, all five anionic AMPs expressed well, with two at unusually high levels (375 and 563 µg/gfw). Furthermore, antimicrobial activity against Staphylococcus epidermidis was an order of magnitude greater (average minimum inhibitory concentration MIC of 0.26 µM) than that typically seen for AMPs expressed in E. coli systems and was associated with the uncleaved fusion peptide. In summary, this study describes a means of expressing AMP fusions in plants in high yield, purified by a simple temperature-shift protocol, resulting in a fusion peptide with high antimicrobial activity and without the need for a peptide cleavage step.

    更新日期:2019-12-05
  • Fast measurement of phosphates and ammonium in fermentation-like media: a feasibility study
    New Biotechnol. (IF 3.739) Pub Date : 2019-11-23
    Katrin Pontius, Giulia Praticò, Flemming H. Larsen, Thomas Skov, Nils Arneborg, Anna Eliasson Lantz, Marta Bevilacqua

    Real-time monitoring of bioprocesses plays a key-role in modern industries, providing new information on full-scale production, thus enabling control of the process and allowing it to run at optimal conditions while minimizing waste. Monitoring of phosphates and ammonium in fermentation processes has a twofold interest: they are important nutrients for living organisms while at the same time constituting environmental nutrient pollutants, for which unnecessary use and disposal must be avoided. In this report, the possibility of simultaneous analysis of phosphates and ammonium in fermentations was verified using spectroscopy-based methods combined with chemometrics to construct calibration models. To achieve this, the models were based on synthetic samples mimicking real fermentation media, providing a dataset where the analytes were completely uncorrelated. Different at-line techniques (mid- and near- infrared spectroscopy, MIR and NIR) were evaluated for their ability to monitor quickly both analytes, in a wide range of concentrations (10-100 mM), in three media of different complexities. Partial Least Squares (PLS) models on MIR spectroscopy gave very good results, with prediction errors lower than 5% for both analytes in all datasets. In contrast, the results for PLS models on NIR spectroscopy were inferior (prediction errors between 3 and 26%) for both analytes, as, in the case of phosphate, it could be demonstrated that the model was based on based on indirect predictions.

    更新日期:2019-11-26
  • Isopropanol production from carbon dioxide in Cupriavidus necator in a pressurized bioreactor
    New Biotechnol. (IF 3.739) Pub Date : 2019-11-12
    Lucile Garrigues, Louna Maignien, Eric Lombard, Jasbir Singh, Stéphane E. Guillouet

    A bioreactor was designed to provide high gas mass transfer to reach cell and product titres in the g L-1 level from CO2 for realistic, laboratory scale, engineered autotrophic strain evaluation. The design was based on independent CO2, H2 and air inputs and the ability to operate at high pressures. The bioreactor configuration and cultivation strategy enabled growth of Cupriavidus necator strains for long periods, to reach over 3 g L-1 dry cell weight. No negative impact of the high pressure was observed on viability of the strains up to more than 4 bar overpressure. The cultivation was then carried out using an engineered isopropanol producing strain; in this case, 3.5 g L-1 isopropanol was obtained from CO2 as the sole carbon source. This is the first reported demonstration of a successful production from engineered bacteria of product in the g L-1 range on CO2, raising the prospect of future development of CO2-based bioprocesses.

    更新日期:2019-11-13
  • Advances in combined enzymatic extraction of ferulic acid from wheat bran
    New Biotechnol. (IF 3.739) Pub Date : 2019-11-12
    Maura Ferri, Anton Happel, Giulio Zanaroli, Marco Bertolini, Stefano Chiesa, Mauro Commisso, Flavia Guzzo, Annalisa Tassoni
    更新日期:2019-11-13
  • Improving the mutagenesis efficiency of the Kunkel method by codon optimization and annealing temperature adjustment
    New Biotechnol. (IF 3.739) Pub Date : 2019-11-11
    Bin Liu, Shuang Long, Jianghai Liu

    The Kunkel method is a widely used site-directed mutagenesis strategy that introduces point mutations by annealing mutation-containing oligonucleotides to single-stranded uracil-containing DNA (dU-ssDNA) templates. The method is fast and inexpensive and has been routinely employed to generate point mutations and multi-site mutations. However, its efficiency for point mutations is highly variable. In this work, codons in both DNA templates and mutagenic oligonucleotides were optimized to lower the GC percentage (GC%) of the complementary regions, and the oligonucleotide length was also extended to reduce the GC difference between upstream and downstream regions. These modifications largely increased the mutation efficiency of single-site mutagenesis. In addition, a multi-stage cooling programme was developed in the annealing step specifically for multi-site mutagenesis, which increased the simultaneous mutation efficiency. The modifications will help in generating antibody libraries by effectively randomizing multiple CDRs simultaneously.

    更新日期:2019-11-13
  • Production of polymers by cyanobacteria grown in wastewater: Current status, challenges and future perspectives
    New Biotechnol. (IF 3.739) Pub Date : 2019-09-18
    Dulce María Arias, Joan García, Enrica Uggetti

    Cyanobacteria are prokaryotic oxygenic phototrophs receiving attention in a wide variety of technological applications such as food and feed supplements and production of valuable polymers. Among these, carbohydrates (e.g. glycogen) and polyhydroxyalkanoates (PHAs) are of increasing interest due to their potential as a biofuel substrate and bioplastics, respectively. However, biofuels and bioplastics from cyanobacteria have seen many years of effort towards commercialization with only limited success. Their main limitation for polymer production is the high cost of the nutrient source; wastewater, as an inexpensive and widely available alternative, may overcome this bottleneck. Though cyanobacteria have demonstrated a capacity to treat wastewater effluents, their cultivation in such a variable environment involves certain challenges of which the chief one is linked to contamination by other species, especially green algae. This would represent a serious drawback during cyanobacterial biomass production and affect further PHA and carbohydrate production. The present study reviews the potential of cyanobacteria to grow in wastewater effluents from different sources. Conditions favoring them in mixed-culture reactors are described, focusing on nutritional and operational aspects. Current advances and future prospects in PHA and carbohydrate production are explored and discussed.

    更新日期:2019-11-08
  • Metabolic processes applied to endangered metal and wood heritage objects: Call a microbial plumber!
    New Biotechnol. (IF 3.739) Pub Date : 2019-11-06
    Edith Joseph, Pilar Junier

    While often considered as harmful for cultural heritage, microorganisms can also be used for its safeguarding. The methods used so far for the conservation-restoration of cultural heritage are often unsatisfactory in terms of efficiency and durability. Inhibitors or complexing agents are also toxic and pose potential threats to human health and to the environment. Microbial-based technologies can provide sustainable solutions for heritage conservation-restoration using ecologically friendly biological treatments. Over the last decades, the development of biological methods and materials has become a significant alternative for the preservation of ancient heritage. Of particular note, microbial metabolisms are exploited to consolidate, clean, stabilize or even protect surfaces of cultural items. Taking advantage of unique properties of microorganisms, reactive corrosion products are extracted or converted into biogenic minerals that provide the treated surfaces with long-term stability. Examples of the techniques proposed include the formation of passivating biogenic layers that can be applied for preservation of metal-based heritage, as well as the development of methods for the preventative removal of iron species from waterlogged wood. This review presents the current advance made in research aiming to preserve copper- and iron-based artefacts, in particular sculptures but also archaeological objects, as well as in the development of a method for the extraction of iron species from waterlogged wood.

    更新日期:2019-11-06
  • A brick in the wall: Discovering a novel mineral component of the biofilm extracellular matrix
    New Biotechnol. (IF 3.739) Pub Date : 2019-11-06
    Alona Keren-Paz, Ilana Kolodkin-Gal

    Multicellular bacterial communities, known as biofilms, have been thought to be held together solely by a self-produced organic extracellular matrix (ECM). However, new evidence for a missed mineral constituent of ECM in both Gram-positive and Gram-negative bacterial species, is accumulating. Study of two phylogenetically distinct bacteria, Bacillus subtilis and Mycobacterium smegmatis, identified a novel mechanism crucial for proper biofilm development and architecture – an active, genetically regulated, production of crystalline calcite. The calcite scaffolds stabilize bacterial biofilms, limit penetration of small molecule solutes such as antibiotics and play a conserved role in the assembly of those complex differentiated multicellular communities. Accumulating evidence suggests tight biological regulation of the formation of these functional minerals. This review discusses the recently discovered structural and functional roles of extracellular minerals in biofilms. It is proposed that it is time for a more complete view of the ECM as a complex combination of organic and nonorganic materials, especially in the light of the possible implications for treatment of biofilm infections.

    更新日期:2019-11-06
  • Model-based analysis of biocatalytic processes and performance of microbioreactors with integrated optical sensors
    New Biotechnol. (IF 3.739) Pub Date : 2019-11-05
    Daria Semenova, Ana C. Fernandes, Juan M. Bolivar, Inês P. Rosinha Grundtvig, Barbara Vadot, Silvia Galvanin, Torsten Mayr, Bernd Nidetzky, Alexandr Zubov, Krist V. Gernaey

    Design and development of scale-down approaches, such as microbioreactor (µBR) technologies with integrated sensors, are an adequate solution for rapid, high-throughput and cost-effective screening of valuable reactions and/or production strains, with considerably reduced use of reagents and generation of waste. A significant challenge in the successful and widespread application of µBRs in biotechnology remains the lack of appropriate software and automated data interpretation of µBR experiments. Here, it is demonstrated how mathematical models can be usedas helpful tools, not only to exploit the capabilities of microfluidic platforms, but also to reveal the critical experimental conditions when monitoring cascade enzymatic reactions. A simplified mechanistic model was developed to describe the enzymatic reaction of glucose oxidase and glucose in the presence of catalase inside a commercial microfluidic platform with integrated oxygen sensor spots. The proposed model allowed an easy and rapid identification of the reaction mechanism, kinetics and limiting factors. The effect of fluid flow and enzyme adsorption inside the microfluidic chip on the optical sensor response and overall monitoring capabilities of the presented platform was evaluated via computational fluid dynamics (CFD) simulations. Remarkably, the model predictions were independently confirmed for µL- and mL- scale experiments. It is expected that the mechanistic models will significantly contribute to the further promotion of µBRs in biocatalysis research and that the overall study will create a framework for screening and evaluation of critical system parameters, including sensor response, operating conditions, experimental and microbioreactor designs.

    更新日期:2019-11-06
  • A compact nanobody-DNAzyme conjugate enables antigen detection and signal amplification
    New Biotechnol. (IF 3.739) Pub Date : 2019-11-01
    Giulio Bernardinelli, Sandra Oloketuyi, Salesia Franziska Werner, Elisa Mazzega, Björn Högberg, Ario de Marco
    更新日期:2019-11-01
  • Corrigendum to "The pre-analytical phase of liquid biopsy" [New Biotechnol. 55, 25 March 2020, 19-29].
    New Biotechnol. (IF 3.739) Pub Date : null
    F Salvianti,S Gelmini,F Costanza,I Mancini,G Sonnati,L Simi,M Pazzagli,P Pinzani

    更新日期:2019-11-01
  • Environmental electroactive consortia as reusable biosensing element for freshwater toxicity monitoring.
    New Biotechnol. (IF 3.739) Pub Date : 2019-09-29
    V Agostino,G Massaglia,M Gerosa,A Sacco,G Saracco,V Margaria,M Quaglio

    The development of tools to monitor water quality is mandatory in a scenario where clean water resources are decreasing. Here, the biosensing capability of an electroactive river sediment consortium was tested towards three model contaminants (glutaraldehyde, nickel(II) and chromium(III)). The proposed biosensor is a small membrane-less single chamber Microbial Fuel Cell (MFC), fabricated by 3D printing. Its semi-continuous mode of operation resulted in long-term current profile stability and reproducibility. A linear trend of response was obtained for glutaraldehyde in a concentration range of 5-1000 ppm. After the recovery of the electroactive consortium activity, the MFC-based biosensors were shown to be sensitive towards Ni(II) and Cr(III), at concentrations above 2 mg L-1. To effectively analyze biosensor response, a novel algorithm was proposed, offering advantages for the realization of energy-saving protocols for MFC-biosensor data transmission. Implementation of the device and method, from laboratory test to real environment, can offer a low cost in situ system for detection of water contaminants.

    更新日期:2019-11-01
  • The future of technologies for personalised medicine.
    New Biotechnol. (IF 3.739) Pub Date : 2012-10-24
    Alison Harvey,Angela Brand,Stephen T Holgate,Lars V Kristiansen,Hans Lehrach,Aarno Palotie,Barbara Prainsack

    Personalised medicine promises prediction, prevention and treatment of illness that is targeted to individuals’ needs. New technologies for detailed biological profiling of individuals at the molecular level have been crucial in initiating the move to personalised medicine; further novel technologies will be necessary if the vision is to become a reality. We will need to develop new technologies to collect and analyse data in a way that is not just linear but integrated (understanding system level functioning) and dynamic (understanding system in flux). Key factors in the development of technologies for personalised medicine are standardisation, integration and harmonisation. For example, the tools and processes for data collection and analysis must be standardised across research sites. Research activity at different sites must be integrated to maximise synergies, and scientific research must be integrated with healthcare to ensure effective translation. There must also be harmonisation between scientific practices in different research sites, between science and healthcare and between science, healthcare and wider society, including the ethical and regulatory frameworks, the prevailing political and cultural ethos and the expectations of patients/citizens.

    更新日期:2019-11-01
  • Tyrosine sulfation: an increasingly recognised post-translational modification of secreted proteins.
    New Biotechnol. (IF 3.739) Pub Date : 2009-08-07
    Martin J Stone,Sara Chuang,Xu Hou,Menachem Shoham,John Z Zhu

    The post-translational sulfation of tyrosine residues occurs in numerous secreted and integral membrane proteins and, in many cases, plays a crucial role in controlling the interactions of these proteins with physiological binding partners as well as invading pathogens. Recent advances in our understanding of protein tyrosine sulfation have come about owing to the cloning of two human tyrosylprotein sulfotransferases (TPST-1 and TPST-2), the development of novel analytical and synthetic methodologies and detailed studies of proteins and peptides containing sulfotyrosine residues. In this article, we describe the TPST enzymes, review the major techniques available for studying the presence, location and function of tyrosine sulfation in proteins and discuss the biological functions and biochemical interactions of several proteins (or protein families) in which tyrosine sulfation influences the protein function. In particular, we describe the detailed evidence supporting the importance of tyrosine sulfation in the cellular adhesion function of P-selectin glycoprotein ligand-1, the leukocyte trafficking and pathogen invasion functions of chemokine receptors and the ligand binding and activation of other G-protein-coupled receptors by complement proteins, phospholipdis and glycoprotein hormones.

    更新日期:2019-11-01
  • Applications of nuclear reprogramming and directed differentiation in vascular regenerative medicine.
    New Biotechnol. (IF 3.739) Pub Date : 2014-07-30
    Amer A Rana,Elizabeth M Callery

    As vertebrates proceed through embryonic development the growing organism cannot survive on diffusion of oxygen and nutrients alone and establishment of vascular system is fundamental for embryonic development to proceed. Dysfunction of the vascular system in adults is at the heart of many disease states such as hypertension and atherosclerosis. In this review we will focus on attempts to generate the key cells of the vascular system, the endothelial and smooth muscle cells, using human embryonic stem cells (hESCs) and human induced pluripotent stem cells (iPSCs). Regardless of their origin, be it embryonic or via somatic cell reprogramming, pluripotent stem cells provide limitlessly self-renewing populations of material suitable for the generation of multi-lineage isogenic vascular cells-types that can be used as tools to study normal cell and tissue biology, model disease states and also as tools for drug screening and future cell therapies.

    更新日期:2019-11-01
  • Reducing acetate excretion from E. coli K-12 by over-expressing the small RNA SgrS.
    New Biotechnol. (IF 3.739) Pub Date : 2011-11-24
    Alejandro Negrete,Nadim Majdalani,Je-Nie Phue,Joseph Shiloach

    When exposed to the nonmetabolized glucose derivative alpha methyl glucoside (αMG), both Escherichia coli K-12 (JM109 and MG1655) and E. coli B (BL21) respond by reducing the concentration of the mRNA of the ptsG gene which is responsible for the biosynthesis of the glucose transporter EIICB(glu). This occurs through the over-expression of the noncoding small RNA SgrS, which interacts specifically with the mRNA of the ptsG gene and prevents its translation. However, when these bacteria are exposed to a glucose concentration of 40 g/L, over-expression of SgrS is observed only in E. coli B (BL21). Unlike E. coli K-12 (JM109 and MG1655), which are affected by high glucose concentration and produce higher levels of acetate, E. coli B (BL21) is not affected. Based on this information, it was assumed that over-expression of SgrS enables E. coli B (BL21) to reduce its acetate excretion by controlling the glucose transport. When SgrS was over-expressed in both E. coli K-12 strains from a multicopy plasmid, it was possible to reduce their acetate excretion levels to those seen in E. coli B. This observation opens a new approach towards controlling bacterial metabolism through the use of noncoding RNA.

    更新日期:2019-11-01
  • In vitro protein expression: an emerging alternative to cell-based approaches.
    New Biotechnol. (IF 3.739) Pub Date : 2010-09-02
    Mingyue He

    Protein expression remains a bottleneck in the production of proteins. Owing to several advantages, cell-free translation is emerging as an alternative to cell-based methods for the generation of proteins. Recent advances have led to many novel applications of cell-free systems in biotechnology, proteomics and fundamental biological research. This special issue of New Biotechnology describes recent advances in cell-free protein expression systems and their applications.

    更新日期:2019-11-01
  • Detection of protein-protein interactions by ribosome display and protein in situ immobilisation.
    New Biotechnol. (IF 3.739) Pub Date : 2009-09-02
    Mingyue He,Hong Liu,Martin Turner,Michael J Taussig

    We describe a method for identification of protein-protein interactions by combining two cell-free protein technologies, namely ribosome display and protein in situ immobilisation. The method requires only PCR fragments as the starting material, the target proteins being made through cell-free protein synthesis, either associated with their encoding mRNA as ribosome complexes or immobilised on a solid surface. The use of ribosome complexes allows identification of interacting protein partners from their attached coding mRNA. To demonstrate the procedures, we have employed the lymphocyte signalling proteins Vav1 and Grb2 and confirmed the interaction between Grb2 and the N-terminal SH3 domain of Vav1. The method has promise for library screening of pairwise protein interactions, down to the analytical level of individual domain or motif mapping.

    更新日期:2019-11-01
  • Microspheres as a vehicle for biomolecule delivery to neural stem cells.
    New Biotechnol. (IF 3.739) Pub Date : 2009-06-16
    Nicole Gennet,Lois M Alexander,Rosario M Sánchez-Martín,Jonathan M Behrendt,Andrew J Sutherland,Joshua M Brickman,Mark Bradley,Meng Li

    Neural stem cells (NSC) are a valuable model system for understanding the intrinsic and extrinsic controls for self-renewal and differentiation choice. They also offer a platform for drug screening and neurotoxicity studies, and hold promise for cell replacement therapies for the treatment of neurodegenerative diseases. Fully exploiting the potential of this experimental tool often requires the manipulation of intrinsic cues of interest using transfection methods, to which NSC are relatively resistant. In this paper, we show that mouse and human NSC readily take up polystyrene-based microspheres which can be loaded with a range of chemical or biological cargoes. This uptake can take place in the undifferentiated stage without affecting NSC proliferation and their capacity to give rise to neurons and glia. We demonstrate that beta-galactosidase-loaded microspheres could be efficiently introduced into NSC with no apparent toxic effect, thus providing proof-of-concept for the use of microspheres as an alternative biomolecule delivery system.

    更新日期:2019-11-01
  • Pre-analytical processes in medical diagnostics: New regulatory requirements and standards.
    New Biotechnol. (IF 3.739) Pub Date : 2019-05-19
    Georges Dagher,Karl-Friedrich Becker,Serena Bonin,Carole Foy,Stefania Gelmini,Mikael Kubista,Penelope Kungl,Uwe Oelmueller,Helen Parkes,Pamela Pinzani,Peter Riegman,Ulrike Schröder,Cornelia Stumptner,Paola Turano,Robert Sjöback,Andrea Wutte,Kurt Zatloukal

    In May 2017, the European In Vitro Diagnostic Regulation (IVDR) entered into force and will apply to in vitro diagnostics from May 26th, 2022. This will have a major impact on the in vitro diagnostics (IVD) industry as all devices falling under the scope of the IVDR will require new or re-certification. It will also affect health institutions developing and using in-house devices. The IVDR also has implications with respect to product performance validation and verification including the pre-analytics of biological samples used by IVD developers and diagnostic service providers. In parallel to the IVDR, a series of standards on pre-analytical sample processing has been published by the International Organization for Standardization (ISO) and the European Committee for Standardization (CEN). These standards describe pre-analytical requirements for various types of analyses in various types of biospecimens. They are of relevance for IVD product developers in the context of (re)certification under the IVDR and to some extent also to devices manufactured and used only within health institutions. This review highlights the background and the rational for the pre-analytical standards. It describes the procedure that leads to these standards, the major implications of the standards and the requirements on pre-analytical workflows. In addition, it discusses the relationship between the standards and the IVDR.

    更新日期:2019-11-01
  • Gene transfection of HEK cells on supermacroporous polyacrylamide monoliths: a comparison of transient and stable recombinant protein expression in perfusion culture.
    New Biotechnol. (IF 3.739) Pub Date : 2009-08-26
    Matthew C Cheeks,Alexander D Edwards,Christopher J Arnot,Nigel K H Slater

    Transient and continuous recombinant protein expression by HEK cells was evaluated in a perfused monolithic bioreactor. Highly porous synthetic cryogel scaffolds (10 ml bed volume) were characterised by scanning electron microscopy and tested as cell substrates. Efficient seeding was achieved (94% inoculum retained, with 91-95% viability). Metabolite monitoring indicated continuous cell growth, and endpoint cell density was estimated by genomic DNA quantification to be 5.2 x 10(8), 1.1 x 10(9) and 3.5 x 10(10) at day 10, 14 and 18. Culture of stably transfected cells allowed continuous production of the Drosophila cytokine Spätzle by the bioreactor at the same rate as in monolayer culture (total 1.2mg at day 18) and this protein was active. In transient transfection experiments more protein was produced per cell compared with monolayer culture. Confocal microscopy confirmed homogenous GFP expression after transient transfection within the bioreactor. Monolithic bioreactors are thus a flexible and powerful tool for manufacturing recombinant proteins.

    更新日期:2019-11-01
  • Plant Biotechnology: Green for Good IV.
    New Biotechnol. (IF 3.739) Pub Date : 2018-05-31
    Petr Tarkowski,Ivo Frébort

    更新日期:2019-11-01
  • Optimized synthesis of 1,3-dioleoyl-2-palmitoylglycerol-rich triacylglycerol via interesterification catalyzed by a lipase from Thermomyces lanuginosus.
    New Biotechnol. (IF 3.739) Pub Date : 2009-11-03
    Jeung Hee Lee,Jeoung Mae Son,Casimir C Akoh,Mee Ree Kim,Ki-Teak Lee

    1,3-Dioleoyl-2-palmitoylglycerol (OPO)-rich human milk fat substitute (HMFS) was synthesized from tripalmitin-rich fraction and ethyl oleate by a lipase-catalyzed interesterification. Response surface methodology was employed to optimize its OPO content and acyl migration with reaction factors - substrate mole ratio of PPP-rich fraction to ethyl oleate (1:4, 1:5 and 1:6), reaction temperature (50, 55 and 60 degrees C) and time (3, 7.5 and 12 hours). The predictive models for OPO content and acyl migration were adequate and reproducible. The OPO content increased with substrate ratio, and decreased with reaction time and temperature, whereas acyl migration increased with temperature and time. The optimal conditions for HMFS synthesis while maximizing OPO content (31.43% OPO) and minimizing acyl migration (6.07%) were predicted at the reaction combination of 50 degrees C, three hours and 5.5 substrate ratio. HMFS was resynthesized under the same condition, and no significant difference between the observed and predicted values was found. Further, the major fatty acid of HMFS was palmitic acid (80.6%) at sn-2 position, and oleic acid (64.9%) at sn-1,3 position.

    更新日期:2019-11-01
  • Fluorescent sperm marking to improve the fight against the pest insect Ceratitis capitata (Wiedemann; Diptera: Tephritidae).
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Francesca Scolari,Marc F Schetelig,Sabrina Bertin,Anna R Malacrida,Giuliano Gasperi,Ernst A Wimmer

    The Sterile Insect Technique (SIT) involving area-wide release of mass-reared and sterilized pest insects has proven successful to reduce, control and eradicate economically important pest species, such as the Mediterranean fruit fly (medfly). For the efficient application, effective monitoring to assess the number and mating success of the released medflies is essential. Here, we report sperm-specific marking systems based on the spermatogenesis-specific Ceratitis capitata beta2-tubulin (Ccbeta2t) promoter. Fluorescent sperm can be isolated from testes or spermathecae. The marking does not cause general disadvantages in preliminary laboratory competitiveness assays. Therefore, transgenic sperm marking could serve as a major improvement for monitoring medfly SIT programs. The use of such harmless transgenic markers will serve as an ideal initial condition to transfer insect transgenesis technology from the laboratory to field applications. Moreover, effective and easily recognizable sperm marking will make novel studies possible on medfly reproductive biology which will help to further improve SIT programs.

    更新日期:2019-11-01
  • Novel technique for scaling up of micropropagated Ruta graveolens shoots using liquid culture systems: a step towards commercialization.
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Renuka Diwan,Nutan Malpathak

    Wide applications of Ruta graveolens L. in pharmaceutical industry has led to increased interest in large-scale plant production, with emphasis on use of in vitro cultures. Earlier reports describe use of in vitro germinated seedlings for raising shoot cultures and not regeneration. There is only a single regeneration protocol of R. graveolens; however, it employs conventional labour intensive techniques deterring automation. The aim of present investigation was to establish a cost effective protocol for large-scale plant production. We report for the first time a one-step protocol with improved regeneration efficiency for multiple shoots induction employing liquid culture systems. Effect of polyamines (putrescine and spermine) on growth and furanocoumarin was studied. Addition of spermine enhanced the number of multiple shoots formed (2.5-fold) and reduced the time taken by half. Spermine addition resulted in 1.47-fold in furanocoumarin production. The selected shoot line, RS2 was successfully scaled up to 5L in culture vessels, with 1.53-fold increase in biomass without affecting the productivity of these cultures. This proves to be a commercially feasible alternative to bioreactors for large-scale biomass and furanocoumarin production.

    更新日期:2019-11-01
  • Shaken helical track bioreactors: Providing oxygen to high-density cultures of mammalian cells at volumes up to 1000 L by surface aeration with air.
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Xiaowei Zhang,Matthieu Stettler,Oscar Reif,Andreas Kocourek,Maria Dejesus,David L Hacker,Florian M Wurm

    A new scalable reactor was developed by applying a novel mixing principle that allows the large-scale cultivation of mammalian cells simply with surface aeration using air owing to increased liquid-gas transfer compared to standard stirred-tank bioreactors. In the cylindrical vessels (50 mL-1500 L) with a helical track attached to the inside wall, the liquid moved upward onto the track as the result of orbital shaking to increase the liquid-gas interface area significantly. This typically resulted in a 5-10-fold improvement in the volumetric mass transfer coefficient (k(L)a). In a 1500-L helical track vessel with a working volume of 1000 L, a k(L)a of 10h(-1) was obtained at a shaking speed of 39 rpm. Cultivations of CHO cells in a shaken 55-L helical track bioreactor resulted in improved cell growth profiles compared to control cultures in standard systems. These results demonstrated the possibility of using these new bioreactors at scales of 1000 L or more.

    更新日期:2019-11-01
  • Space and time-resolved gene expression experiments on cultured mammalian cells by a single-cell electroporation microarray.
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    S Vassanelli,L Bandiera,M Borgo,G Cellere,L Santoni,C Bersani,M Salamon,M Zaccolo,L Lorenzelli,S Girardi,M Maschietto,M Dal Maschio,A Paccagnella

    Single-cell experiments represent the next frontier for biochemical and gene expression research. Although bulk-scale methods averaging populations of cells have been traditionally used to investigate cellular behavior, they mask individual cell features and can lead to misleading or insufficient biological results. We report on a single-cell electroporation microarray enabling the transfection of pre-selected individual cells at different sites within the same culture (space-resolved), at arbitrarily chosen time points and even sequentially to the same cells (time-resolved). Delivery of impermeant molecules by single-cell electroporation was first proven to be finely tunable by acting on the electroporation protocol and then optimized for transfection of nucleic acids into Chinese Hamster Ovary (CHO-K1) cells. We focused on DNA oligonucleotides (ODNs), short interfering RNAs (siRNAs), and DNA plasmid vectors, thus providing a versatile and easy-to-use platform for time-resolved gene expression experiments in single mammalian cells.

    更新日期:2019-11-01
  • SRP and Sec pathway leader peptides for antibody phage display and antibody fragment production in E. coli.
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Holger Thie,Thomas Schirrmann,Matthias Paschke,Stefan Dübel,Michael Hust

    Antibody phage display is a key technology for the generation of recombinant (human) antibodies for research, diagnostics and therapy. Most antibody fragments can only be folded correctly in the oxidizing environment of the periplasm of Escherichia coli. A multitude of leader peptides has been used for secretion of antibody::pIII fusion proteins into the periplasm, but a systematic study of their impact on the performance of antibody phage display systems has not been reported so far. In this work we have analysed the influence of various leader peptides on antibody phage display efficiency and production yields of soluble antibody fragments. Four leader peptides using the Sec pathway (PelB, OmpA, PhoA and pIII) and three using the SRP pathway (DsbA, TorT and TolB) were compared. Both pathways are compatible with antibody phage display and the production of soluble antibody fragments. The applicability of the SRP pathway to antibody phage display and the production of functional scFvs is shown here for the first time.

    更新日期:2019-11-01
  • Ligation-based molecular tools for lab-on-a-chip devices.
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Jonas Melin,Jonas Jarvius,Chatarina Larsson,Ola Söderberg,Ulf Landegren,Mats Nilsson

    Molecular diagnostics can offer early detection of disease, improved diagnostic accuracy, and qualified follow-up. Moreover, the use of microfluidic devices can in principle render these analyses quickly and user-friendly, placing them within the reach of the general practitioner and maybe even in households. However, the progress launching such devices has been limited so far. We propose that an important limiting factor has been the difficulty of establishing molecular assays suitable for microfabricated formats. The assays should be capable of monitoring a wide range of molecules, including genomic DNA, RNA and proteins with secondary modifications and interaction partners, and they must exhibit excellent sensitivity and specificity. We discuss these problems and describe a series of molecular tools that may present new opportunities for lab-on-a-chip devices at the point-of-care.

    更新日期:2019-11-01
  • Company profile: Spider stories.
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Andrea Giuliani

    SpiderBiotech is a biotech company that has carried out extensive research and development on peptide-based anti-infectives, with five people involved in R&D activities and a strong network of industrial and academic partners experienced in the field of anti-infectives. SpiderBiotech has also created a proprietary library of bioactive peptides and lipopeptides (both linear and dendrimeric) active against bacterial and viral infections. At the moment they have two ongoing projects: the most advanced is focused on the development of a panel of peptide based antibiotics. The second project is related to novel antiviral drugs to treat acyclovir resistant Herpes virus infections.

    更新日期:2019-11-01
  • Challenging lifelong learning in biotechnology.
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Simone Maccaferri

    更新日期:2019-11-01
  • Life on top-working@ the Wellcome Trust Centre for Human Genetics: Scotland Yard for DNA detectives.
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Sreeram V Ramagopalan

    The Wellcome Trust Centre for Human Genetics (WTCHG) was established in 1994 to undertake research into the genetic basis of common diseases. Since June 1999 the centre has been located in the Henry Wellcome Building of Genomic Medicine, University of Oxford. The scientific objective of the centre is to explore all aspects of the genetic susceptibility of disease including the localisation of genes involved in common diseases, characterization of the variants responsible for susceptibility, the understanding of how these DNA variants may contribute to risk of disease in the population and finally, the understanding of how such genetic factors contribute biologically to a disease process. The centre houses multidisciplinary research teams in human genetics, functional genomics, bioinformatics, statistical genetics and structural biology.

    更新日期:2019-11-01
  • Interview with Philippe Ory of the EPFL (Ecole Polytechnique Federale de Lausanne) Career Centre. Interviewed by Debora Keller.
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Philippe Ory

    EPFL's Career Centre was created in 2007 with the mission to be a bridge between EPFL's young graduates and industry, in order to facilitate the transition to active scientific life. Through courses, workshops and personalised advice, they help graduates to set up their application documents (CV, motivation letter), prepare for job interviews and manage their careers. The Centre also offers its services to companies by organising on-campus recruitment days, actively searching for fitting profiles or posting or mailing job adverts to the EPFL graduate community. The Career Centre's goal is to be the platform for the EPFL graduates to build their careers.

    更新日期:2019-11-01
  • Interview with professor Fotis C. Kafatos, president of the European Research Council. Interviewed by Francesco Lescai.
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Fotis C Kafatos

    The European Research Council (ERC) is the first European funding body set up to support investigator-driven frontier research. Its main aim is to stimulate scientific excellence by supporting and encouraging the very best, truly creative scientists, scholars and engineers to be adventurous and take risks in their research. The scientists should go beyond the established frontiers of knowledge and the boundaries of disciplines. Being 'investigator-driven', or 'bottom-up', in nature, the ERC approach allows researchers to identify new opportunities and directions in any field of research. By challenging Europe's brightest minds, the ERC expects to bring about new and unpredictable scientific and technological discoveries-the kind that can form the basis of new industries, markets and broader social innovations of the future. Ultimately, the ERC aims to make the European research base more prepared to respond to the needs of a knowledge-based society and provide Europe with the capabilities in frontier research necessary to meet global challenges.

    更新日期:2019-11-01
  • The Young European Biotech Network (YEBN).
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Francesco Lescai

    更新日期:2019-11-01
  • EAGLES report on HIV/AIDS research.
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Robert Walgate,Jens Degett,

    更新日期:2019-11-01
  • Life sciences must focus on the poor.
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Jens Degett,David McConnell

    Following Serageldin, the members of EAGLES believe that the solution to the great world problems 'requires our joint commitment as scientists to work for the benefit of the entire human family, not just the privileged minority who are lucky enough to live in the most advanced industrial societies. These tasks are enormous. But the longest journey starts with a single step. So let us start. If not us, who? If not now, when?' Now is the time for European life sciences to focus their attention on the most pressing problems facing humanity today and accept the fact that these problems are not in Europe.

    更新日期:2019-11-01
  • Innovation in ScanBalt BioRegion.
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Peter Frank,Elise Kvarnström,Frank Graage,Wolfgang Blank,Anna Pytko,Tuula Palmen,Andrew Munk,Mairita Coneva,Jaanus Pikani

    更新日期:2019-11-01
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  • The ESF's EuroBioFund: sustainable research financing? Start with networking!
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Esther Thole,Wouter Spek

    EuroBioFund was set up by the European Science Foundation in 2006, with financial support from the European Commission to catalyse the development of large scale pan-European life sciences research programmes by engaging those involved in planning and funding research.

    更新日期:2019-11-01
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  • Biofuels are dead: long live biofuels(?) - Part one.
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Andrew Moore

    Beleaguered by criticisms, and abused by politicians for ecological target-setting, biofuels are in their darkest hour. But their bringing to trial should remind us - yet again - of something else: the highly questionable sustainability of most of modern agriculture. Is this the end of biofuels? Probably not, but it is certainly the end of a cheap solution to the problem of sustainable portable fuels. Part one of this two-part article focuses on the political and agricultural dimensions of the topic.

    更新日期:2019-11-01
  • Healthy human ageing.
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Brian F C Clark

    更新日期:2019-11-01
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  • New Biotechnology and the European Federation of Biotechnology.
    New Biotechnol. (IF 3.739) Pub Date : 2008-05-28
    Michael J Taussig,Christian Suojanen,Brian F C Clark

    更新日期:2019-11-01
  • Production of high amounts of 3-hydroxypropionaldehyde from glycerol by Lactobacillus reuteri with strongly increased biocatalyst lifetime and productivity.
    New Biotechnol. (IF 3.739) Pub Date : 2011-07-07
    Hendrik Krauter,Thomas Willke,Klaus-Dieter Vorlop

    3-hydroxypropionaldehyde (3HPA) is a promising versatile substance derived from the renewable feedstock glycerol. It is a product of glycerol metabolism in Lactobacillus reuteri. Because of toxic effects, the biotechnological production is poor. In this work the biocatalyst lifetime and product formation could be drastically increased. In the established two-step process already applied, cells are grown in the first step under anaerobic conditions, and in the second step the immobilised or suspended biocatalyst is used for 3HPA-production under strict anaerobic conditions. In the first step it was possible to reach a biomass concentration of 5.5g CDW/L (OD(600)≈23.4). In the second step, normally, 3HPA accumulates to a toxic concentration and the reaction stops in less than 60min because of the interaction of 3HPA with cell components. To prevent this, the toxic product is bound to the newly found scavenger carbohydrazide to form the hydrazone. For the first time it was possible to recycle the immobilised biocatalyst for at least ten cycles (overall life time>33hours) in a repeated batch biotransformation with an overall production of 67g 3HPA. The optimal pH-value was between 6.8 and 7.2 at an optimal temperature of 40-45°C. In a single batch biotransformation with suspended resting cells it was possible to produce 150g/L 3HPA as carbohydrazone at an overall productivity of 10.7gL(-1)hours(-1). In a single fed-batch biotransformation at 45°C 138g/L glycerol was converted into 108g/L 3HPA with an overall productivity of 21.6gL(-1)hours(-1). This is the highest 3HPA concentration and productivities reported so far for the microbial production of 3HPA from glycerol.

    更新日期:2019-11-01
  • Optimization of Brassica napus (canola) explant regeneration for genetic transformation.
    New Biotechnol. (IF 3.739) Pub Date : 2011-07-05
    Priti Maheshwari,Gopalan Selvaraj,Igor Kovalchuk

    Brassica napus (canola) is the second largest oilseed crop in the world. It is among the first crops to be genetically transformed, and genetically modified cultivars are in commercial production at very significant levels. Despite the early lead with respect to transgenesis, there remain cultivars that are recalcitrant to transformation. To address this, we have conducted an elaborate investigation of the conditions for regenerating shoots from hypocotyl explants from four genetic lines: Invigor 5020, Westar and Topas as well as a microspore culture derived line of Topas (Line 4079). We analyzed the effect of hormonal combinations in regeneration medium, donor plant age and explant type on the regeneration capacity of these plants. The analysis showed that hypocotyls of eight-day-old seedlings grown on media supplemented with 1mg/L dinitrophenylhydrazine (2,4-D) produced the most shoots. Globular somatic embryos emerged following two weeks of 2,4-D treatment. When transferred to the medium containing 5mg/L benzyladenine (BA), approximately 82% of embryos produced shoots within six weeks. Invigor plants were shown to regenerate more efficiently than Topas; the number of plantlets regenerated from Invigor was approximately 40-50% more as compared to Topas or Line 4079. When hypocotyl explants were co-cultivated with the Agrobacterium strain GV3101 harboring a binary vector carrying a firefly luciferase reporter gene (LUC), significant numbers of plantlets were LUC-positive in a luciferase assay. Frequency of such plants were: Invigor 5020 (54.2 ± 2.5%), Westar (53.7 ± 5.3), Topas (16.0 ± 0.24) and Line 4079 (13.4 ± 4).

    更新日期:2019-11-01
  • Granular activated carbon based microbial fuel cell for simultaneous decolorization of real dye wastewater and electricity generation.
    New Biotechnol. (IF 3.739) Pub Date : 2011-07-02
    Shafeer Kalathil,Jintae Lee,Moo Hwan Cho

    Decolorization of dye wastewater before discharge is pivotal because of its immense color and toxicities. In this study, a granular activated carbon based microbial fuel cell (GACB-MFC) was used without using any expensive materials like Nafion membrane and platinum catalyst for simultaneous decolorization of real dye wastewater and bioelectricity generation. After 48 hours of GACB-MFC operation, 73% color was removed at anode and 77% color was removed at cathode. COD removal was 71% at the anode and 76% at the cathode after 48 hours. Toxicity measurements showed that cathode effluent was almost nontoxic after 24 hours. The anode effluent was threefold less toxic compared to original dye wastewater after 48 hours. The GACB-MFC produced a power density of 1.7 W/m(3) with an open circuit voltage 0.45 V. One of the advantages of the GACB-MFC system is that pH was automatically adjusted from 12.4 to 7.2 and 8.0 at the anode and cathode during 48 hours operation.

    更新日期:2019-11-01
  • Biotransformation of the organochlorine pesticide trans-chlordane by wood-rot fungi.
    New Biotechnol. (IF 3.739) Pub Date : 2011-07-02
    Pengfei Xiao,Toshio Mori,Ryuichiro Kondo

    There is very limited information on the biotransformation of organochlorine pesticide chlordane by microorganisms, and no systematic study on the metabolic products and pathways for chlordane transformation by wood-rot fungi has been conducted. In this study, trans-chlordane was metabolized with the wood-rot fungi species Phlebia lindtneri, Phlebia brevispora and Phlebia aurea, which are capable of degrading polychlorinated dibenzo-p-dioxin and heptachlor epoxide. At the end of 42 days of incubation, over 50% of trans-chlordane was degraded by the fungal treatments in pure cultures. These fungi transformed trans-chlordane to at least eleven metabolites including a large amount of hydroxylated products such as 3-hydroxychlordane, chlordene chlorohydrin, heptachlor diol, monohydroxychlordene and dihydroxychlordene. P. lindtneri particularly can metabolize oxychlordane, a recalcitrant epoxide product of chlordane, into a hydroxylated product through substitution of chlorine atom by hydroxyl group. The present results suggest that hydroxylation reactions play an important role in the metabolism of trans-chlordane by these Phlebia species. Additionally, transformation of trans-chlordane and production of hydroxylated metabolites were efficiently inhibited by the addition of cytochrome P450 inhibitors, piperonyl butoxide and 1-aminobenzotriazole, demonstrating that fungal cytochrome P450 enzymes are involved in some steps of trans-chlordane metabolism, particularly in the hydroxylation process.

    更新日期:2019-11-01
  • Effect of 2,4,6-trichlorophenol on the microbial activity of adapted anaerobic granular sludge bioaugmented with Desulfitobacterium strains.
    New Biotechnol. (IF 3.739) Pub Date : 2011-07-02
    D Puyol,A F Mohedano,J J Rodriguez,J L Sanz

    The anaerobic degradation of 2,4,6-trichlorophenol (246TCP) has been studied in batch experiments. Granular sludges previously acclimated to 2,4-dichlorophenol (24DCP) and then adapted to at a load of 330 μM 246TCPd(-1) in two expanded granular sludge bed (EGSB) reactors were used. One of the reactors had been bioaugmented with Desulfitobacterium strains whereas the other served as control. 246TCP was tested at concentrations between 250 and 760 μM. The study focused on the fate of both fermentation products and chlorophenols derived from dechlorination of 246TCP. This compound mainly affected the biodegradation of acetate and propionate, which were inhibited at 246TCP concentrations above 380 μM. Lactate and ethanol were also accumulated at 760 μM 246TCP. Methanogenesis was strongly inhibited at 246TCP concentrations higher than 380 μM. A diauxic production of methane was observed, which can be described by a kinetic model in which acetoclastic methanogenesis was inhibited, whereas hydrogenotrophic methanogenesis was hardly affected by 246TCP. The similarity of the kinetic parameters obtained for the control and the bioaugmented sludges (K(i)=175-200 μM 246TCP and n=7) suggests that methanogenesis is not affected by the bioaugmentation. Moreover, the 246TCP dechlorination occurred mainly at ortho position, successively generating 24DCP and 4-chlorophenol (4CP), which was identified as final product. The bioaugmentation does not significantly improve the anaerobic biodegradation of 246TCP. It has been shown that the active biomass is capable of bioaccumulating 246TCP and products from dechlorination, which are subsequently excreted to the bulk medium when the biomass becomes active again. A kinetic model is proposed which simultaneously explains 246TCP and 24DCP reductive dechlorinations and includes the 246TCP bioaccumulation. The values of the kinetic parameters for 246TCP dechlorination were not affected by bioaugmentation (V(max)=5.3 and 5.1 μM h(-1) and K(s)=5.8 and 13.1 μM for control and bioaugmented sludges, respectively).

    更新日期:2019-11-01
  • Influence of temperature, pH and dissolved oxygen concentration on enhanced biological phosphorus removal under strictly aerobic conditions.
    New Biotechnol. (IF 3.739) Pub Date : 2011-07-02
    Tadashi Nittami,Hiroshi Oi,Kanji Matsumoto,Robert J Seviour

    Previous research has suggested that enhanced biological phosphorus removal (EBPR) from wastewater can be achieved under continuous aerobic conditions over the short term. However, little is known how environmental conditions might affect aerobic EBPR performance. Consequently we have investigated the impact of temperature, pH and dissolved oxygen (DO) concentrations on EBPR performance under strictly aerobic conditions. A sequencing batch reactor (SBR) was operated for 108 days on a six-hour cycle (four cycles a day). The SBR ran under alternating anaerobic-aerobic conditions as standard and then operated under strictly aerobic conditions for one cycle every three or four days. SBR operational temperature (10, 15, 20, 25 and 30°C), pH (6, 7, 8 and 9) and DO concentration (0.5, 2.0 and 3.5mg/L) were changed consecutively during the aerobic cycle. Recorded increases in mixed liquor phosphorus (P) concentrations during aerobic carbon source uptake (P release) were affected by the biomass P content rather than the imposed changes in the operational conditions. Thus, P release levels increased with biomass P content. By contrast, subsequent aerobic P assimilation (P uptake) levels were both affected by changes in operational temperature and pH, and peaked at 20-25°C and pH 7-8. Highest P uptake detected under these SBR operating conditions was 15.4 mg Pg-MLSS(-1) (at 25°C, pH 7 and DO 2.0mg/L). The ability of the community for linked aerobic P release and P uptake required the presence of acetate in the medium, a finding which differs from previous data, where these are reported to occur in the absence of any exogenous carbon source. Fluorescence in situ hybridization was performed on samples collected from the SBR, and Candidatus 'Accumulibacter phosphatis' cells were detected with PAOmix probes through the operational periods. Thus, Candidatus 'Accumulibacter phosphatis' seemed to perform P removal in the SBR as shown in previous studies on P removal under strictly aerobic conditions.

    更新日期:2019-11-01
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