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At least 253 records · Page 14Linked to original sources
PubMed1992

The implantation of life on Mars: feasibility and motivation.

Environmental conditions on Mars are extremely hostile, and would be destructive to any organisms which might arrive there unprotected to-day. However, it is a biocompatible planet. Its unalterable astrophysical parameters would allow the maintenance of a much thicker, warmer carbon dioxide atmosphere than that which currently exists. Though very cold (averaging about -60 degrees C), highly oxidizing and desiccated, Mars may possess substantial quantities of the materials needed to support life--in particular, water and carbon dioxide. A general scenario for implanting life on Mars would include three main phases: (1) robotic and human exploration to determine whether sufficiently large and accessible volatile inventories are available; (2) planetary engineering designed to warm the planet, release liquid water and produce a thick carbon dioxide atmosphere; and (3) if no indigenous Martian organisms emerge as liquid water becomes available, a program of biological engineering designed to construct and implant pioneering microbial communities able to proliferate in the newly clement, though still anaerobic, Martian environment. The process of establishing an ecosystem, or biosphere, on a lifeless planet is best termed 'ecopoiesis.' This new word, derived from Greek, means 'the making of an abode for life.' It is by no means clear whether ecopoiesis on Mars is scientifically possible or technologically achievable. Thus we urge that it be one of the objectives of space research during the next century to assess the feasibility of ecopoiesis on Mars.

Atmosphere↗
PubMed2005

Model of central and trimethylammonium metabolism for optimizing L-carnitine production by E. coli.

The application of metabolic engineering principles to the rational design of microbial production processes crucially depends on the ability to make quantitative descriptions of the systemic ability of the central carbon metabolism to redirect fluxes to the product-forming pathways. The aim of this work was to further our understanding of the steps controlling the biotransformation of trimethylammonium compounds into L-carnitine by Escherichia coli. Despite the importance of L-carnitine production processes, development of a model of the central carbon metabolism linked to the secondary carnitine metabolism of E. coli has been severely hampered by the lack of stoichiometric information on the metabolic reactions taking place in the carnitine metabolism. Here we present the design and experimental validation of a model which, for the first time, links the carnitine metabolism with the reactions of glycolysis, the tricarboxylic acid cycle and the pentose-phosphate pathway. The results demonstrate a need for a high production rate of ATP to be devoted to the biotransformation process. The results demonstrate that ATP is used up in a futile cycle, since both trimethylammonium compound carriers CaiT and ProU operate simultaneously. To improve the biotransformation process, resting processes as well as CaiT or ProU knock out mutants would yield a more efficient system for producing L-carnitine from crotonobetaine or D-carnitine.

Carnitine↗
PubMed2001

Applications of DNA microarrays in microbial systems.

DNA microarray technology allows a parallel analysis of RNA abundance and DNA homology for thousands of genes in a single experiment. Over the past few years, this powerful technology has been used to explore transcriptional profiles and genome differences for a variety of microorganisms, greatly facilitating our understanding of microbial metabolism. With the increasing availability of complete microbial genomes, DNA microarrays are becoming a common tool in many areas of microbial research, including microbial physiology, pathogenesis, epidemiology, ecology, phylogeny, pathway engineering and fermentation optimization.

DNA, Bacterial↗
PubMed2000

Environmental biotechnology.

There is an increasing interest in environmental biotechnology owing to a worldwide need to feed the world's growing population and to maintain clean soil, air and water. The major technological developments are in plant and microbial biology. Plants can be more readily engineered for resistances that enhance yield or produce new products whereas microorganisms are exploited for their catalytic diversity and ease of genetic engineering.

Biotechnology↗
PubMed1999

The domain-specific probe EUB338 is insufficient for the detection of all Bacteria: development and evaluation of a more comprehensive probe set.

In situ hybridization with rRNA-targeted oligonucleotide probes has become a widely applied tool for direct analysis of microbial population structures of complex natural and engineered systems. In such studies probe EUB338 (AMANN et al., 1990) is routinely used to quantify members of the domain Bacteria with a sufficiently high cellular ribosome content. Recent reevaluations of probe EUB338 coverage based on all publicly available 16S rRNA sequences, however, indicated that important bacterial phyla, most notably the Planctomycetales and Verrucomicrobia, are missed by this probe. We therefore designed and evaluated two supplementary versions (EUB338-II and EUB338-III) of probe EUB338 for in situ detection of most of those phyla not detected with probe EUB338. In situ dissociation curves with target and non-target organisms were recorded under increasing stringency to optimize hybridization conditions. For that purpose a digital image software routine was developed. In situ hybridization of a complex biofilm community with the three EUB338 probes demonstrated the presence of significant numbers of probe EUB338-II and EUB338-III target organisms. The application of EUB338, EUB338-II and EUB338-III should allow a more accurate quantification of members of the domain Bacteria in future molecular ecological studies.

Bacteria↗
PubMed2001

Biogeochemical and ecological considerations in designing wetland treatment systems in post-mining landscapes.

Although wetlands have gained acceptance as important components of ecosystems in post-mining landscapes in the past decade, their roles in contaminant retention/removal have not been well integrated into the designing of restoration programs. This paper describes the integration of sediment microbial activities and natural precipitation processes, along with approaches to defining the contaminant load from the mine wastes. The contaminant removal rates, which can be expected by a wetland sediment, are summarized and how they need to be reflected in the wetland size required, and the carbon supply which is needed. Contaminant loading from mining wastes can be balanced by wetland ecological processes, including wetland primary production and microbial mineralization in the sediment. This ecological engineering approach is demonstrated using case studies on hard-rock mining waste in Canada.

Canada↗
PubMedDate not supplied

Affinity purification and characterization of anti-Tac(Fv)-C3-PE38KDEL: A highly potent cytotoxic agent specific to cells bearing IL-2 receptors.

A chimeric, single chain antibody fused immunotoxin, denoted anti-Tac(Fv)-C3-PE38KDEL, was engineered and expressed in Escherichia coli. The microbially expressed anti-Tac(Fv)-C3-PE38KDEL was solubilized from inclusion bodies using guanidine hydrochloride, and subsequently refolded in a redox buffer via thiol/disulfide exchange. The recombinant immunotoxin from the crude extract was purified employing receptor-affinity chromatography, which is based upon biological function and involved the immobilized p55 subunit of human IL-2 receptor. The cytotoxic activity of this immunotoxin was measured by the IL-2 dependent phytohemagglutinin (PHA) blast proliferation inhibition and HUT-102 protein synthesis inhibition assays, in which the IC50 values were 41.5 and 0.8 pM, respectively. The biochemical homogeneity and authenticity of the purified material were determined by gel permeation chromatography, amino acid composition and N-terminal sequence analyses, SDS-PAGE, isoelectric focusing, and Western blotting. The receptor-affinity-purified immunotoxin was shown to be highly effective in specifically killing cells bearing IL-2 receptors. Anti-Tac(Fv)-C3-PE38KDEL is a powerful immunosuppressant which may be a potentially useful therapeutic agent in the prevention of allograft rejection and in the treatment of autoimmune diseases. Another anticipated application of this fusion protein is as a chemotoxin in the treatment of some forms of cancer.

ADP Ribose Transferases↗
PubMed1992

Sequence-specific cleavage of protein fusions using a recombinant Neisseria type 2 IgA protease.

Sequence-specific enzymatic cleavage of protein fusions is an important application in recombinant protein technology. We have used the Neisseria type 2 IgA protease (EC 3.4.24.13), produced and secreted by Escherichia coli host cells, for efficiently processing polypeptides at authentic or engineered target sites. In different substrates, the microbial protease specifically cleaves the peptide bond distal to the second Pro residue of the sequence Yaa-Pro-/-Xaa-Pro, where Yaa stands for Pro (or rarely for Pro in combination with Ala, Gly or Thr) and Xaa stands for Thr, Ser or Ala. Highly specific proteolysis has been obtained not only with soluble and purified protein fusions but also with insoluble aggregates derived from cytoplasmic inclusion bodies. The sequence-specificity and simple production of the recombinant IgA protease make it a versatile tool for the in vitro processing of recombinant proteins.

Amino Acid Sequence↗
PubMed2006

Inhibitors of NF-kappaB signaling: 785 and counting.

Nuclear factor kappa B (NF-kappaB) transcription factors regulate several important physiological processes, including inflammation and immune responses, cell growth, apoptosis, and the expression of certain viral genes. Therefore, the NF-kappaB signaling pathway has also provided a focus for pharmacological intervention, primarily in situations of chronic inflammation or in cancer, where the pathway is often constitutively active and plays a key role in the disease. Now that many of the molecular details of the NF-kappaB pathway are known, it is clear that modulators of this pathway can act at several levels. As described herein, over 750 inhibitors of the NF-kappaB pathway have been identified, including a variety of natural and synthetic molecules. These compounds include antioxidants, peptides, small RNA/DNA, microbial and viral proteins, small molecules, and engineered dominant-negative or constitutively active polypeptides. Several of these molecules act as general inhibitors of NF-kappaB induction, whereas others inhibit specific pathways of induction. In addition, some compounds appear to target multiple steps in the NF-kappaB pathway. Compounds designed as specific NF-kappaB inhibitors are not yet in clinical use, but they are likely to be developed as treatments for certain cancers and neurodegenerative and inflammatory diseases. Moreover, the therapeutic and preventative effects of many natural products may, at least in part, be due to their ability to inhibit NF-kappaB.

Animals↗
PubMed2003

Dispelling the myths--biocatalysis in industrial synthesis.

Biocatalysis has emerged as an important tool in the industrial synthesis of bulk chemicals, pharmaceutical and agrochemical intermediates, active pharmaceuticals, and food ingredients. However, the number and diversity of the applications are modest, perhaps in part because of perceived or real limitations of biocatalysts, such as limited enzyme availability, substrate scope, and operational stability. Recent scientific breakthroughs in genomics, directed enzyme evolution, and the exploitation of biodiversity should help to overcome these limitations. As a result, we expect many new industrial applications of biocatalysis to be realized, from single-step enzymatic conversions to customized multistep microbial synthesis by means of metabolic pathway engineering.

Biotechnology↗
PubMed1983

[Can extrachromosomal genetic elements be recognized as inventions?].

The legal bases for acknowledgement of plasmids (vectors) as inventions, in particular within the classes of the "substance" and "microbial strain" are discussed. There is a definite similarity in a number of features between phages recognized as inventions and plasmids. Therefore, in principle their acknowledgement as inventions may be considered possible, since it raises no new problems as those occurring in legal protection of microbial strains. At the same time there is no pressing need in acknowledgement of plasmids (vectors) as inventions and their legal protection should be rather of the prestige character.

Extrachromosomal Inheritance↗
PubMed1999

Biosynthesis and regulation of microbial polyunsaturated fatty acid production.

Growing interest in polyunsaturated fatty acid (PUFA) applications in various fields coupled with their significance in health and dietary requirements has focused attention on the provision of suitable sources of these compounds. Isolation of highly efficient oleaginous microorganisms has led to the development of fermentation technologies as an alternative to agricultural and animal processes. Particularly active in PUFA synthesis are the Zygomycetes fungi and certain microalgae. Emphasis is placed on increasing the product value by employing new biotechnological strategies (e.g. mutation techniques, molecular engineering and biotransformations) which allow the regulation of microbial PUFA formation with satisfactory yield in order to be competitive with other sources. Comparative successes in fungal PUFA production demonstrate microbial potential to synthesize high-value oils and provide the main stimulus for their applications.

Journal Article↗
PubMed2006

Food-processing enzymes from recombinant microorganisms--a review.

Enzymes are commonly used in food processing and in the production of food ingredients. Enzymes traditionally isolated from culturable microorganisms, plants, and mammalian tissues are often not well-adapted to the conditions used in modern food production methods. The use of recombinant DNA technology has made it possible to manufacture novel enzymes suitable for specific food-processing conditions. Such enzymes may be discovered by screening microorganisms sampled from diverse environments or developed by modification of known enzymes using modern methods of protein engineering or molecular evolution. As a result, several important food-processing enzymes such as amylases and lipases with properties tailored to particular food applications have become available. Another important achievement is improvement of microbial production strains. For example, several microbial strains recently developed for enzyme production have been engineered to increase enzyme yield by deleting native genes encoding extracellular proteases. Moreover, certain fungal production strains have been modified to reduce or eliminate their potential for production of toxic secondary metabolites. In this article, we discuss the safety of microorganisms used as hosts for enzyme-encoding genes, the construction of recombinant production strains, and methods of improving enzyme properties. We also briefly describe the manufacture and safety assessment of enzyme preparations and summarize options for submitting information on enzyme preparations to the US Food and Drug Administration.

Animals↗
PubMed2001

Metal and radionuclide bioremediation: issues, considerations and potentials.

Recent demonstrations of the removal and immobilization of inorganic contaminants by microbial transformations, sorption and mineralization show the potential of both natural and engineered microbes as bioremedial tools. Demonstrations of microbe-mediated mineral formation in biofilms implicate this mode of microbial life in geological evolution and remediation of inorganic contaminants.

Bacteria↗
PubMed2003

Engineering of secondary metabolite pathways.

Nature produces an astonishing wealth of secondary metabolites with important biological functions. To access this diversity of structurally complex chemical compounds for industrial and biomedical applications, cells have been engineered to produce higher levels and/or novel compounds that were previously inaccessible. Recent examples of metabolic and combinatorial engineering illustrate different strategies for the production of secondary metabolites in microbial cells.

Biopolymers↗
PubMed2006

Anti-tumor immunotherapy via antigen delivery from a live attenuated genetically engineered Pseudomonas aeruginosa type III secretion system-based vector.

Immunotherapy requiring an efficient T lymphocyte response is initiated by antigen delivery to antigen-presenting cells. Several studies have assessed the efficiency of various antigen loading procedures, including microbial vectors. Here a live strain of Pseudomonas aeruginosa was engineered to translocate a recombinant antigenic protein into mammalian cells via the type III secretion system, a bacterial device translocating effector proteins into host cells. Optimization of the vector included virulence attenuation and determination of the N-terminal sequence allowing translocation of fused antigens into cells. In vitro delivery of an ovalbumin fragment by the bacterial vector into dendritic cells induced the activation of ovalbumin-specific CD8(+) T lymphocytes. Mice injected with the ovalbumin-delivering vector developed ovalbumin-specific CD8(+) T lymphocytes and were resistant to a subsequent challenge with an ovalbumin-expressing melanoma. Moreover, in a curative assay, injection of the vaccine vector 5 and 12 days after tumor implantation led to a complete cure in five of six animals. These results highlight the utility of type III secretion system-based vectors for anti-tumor immunotherapy.

Animals↗
PubMed1992

Adaptation of model genetically engineered microorganisms to lake water: growth rate enhancements and plasmid loss.

When a genetically engineered microorganism (GEM) is released into a natural ecosystem, its survival, and hence its potential environmental impact, depends on its genetic stability and potential for growth under highly oligotrophic conditions. In this study, we compared plasmid stability and potential for growth on low concentrations of organic nutrients of strains of Pseudomonas putida serving as model GEMs. Plasmid-free and plasmid-bearing (NAH7) prototrophic isogenic strains and two amino-acid auxotrophs, all containing antibiotic resistance markers, were held physically separate from but in chemical contact with lake water containing the natural bacterium-sized microbial populations. Cells were reisolated at intervals over a 2-month period to determine the percent retaining the plasmid and the specific growth rate on various media. Plasmid stability in lake water was strongly strain specific; the NAH7 plasmid was stably maintained by the prototrophic strain for the duration of the test but was lost within 24 h by both of the auxotrophs. Specific growth rates of reisolates, compared with those of the corresponding non-lake water-exposed strains (i.e., parental strains), were not different when measured in rich medium (Luria-Bertani broth). However, specific growth rates were 42, 55, and 63% higher in reisolates of auxotrophs and the plasmid-free prototroph, respectively, when measured in 10-fold-diluted medium after exposure of 15 days or longer to lake water. Moreover, lake water-exposed strains grew actively when reintroduced into sterile lake water (28- to 33-fold increase in numbers over 7 days), while the corresponding unadapted parental strains exhibited no growth over the same period.(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptation, Biological↗
PubMed1998

Genetic manipulation of antitumor-agent biosynthesis to produce novel drugs.

Current methods of obtaining novel drugs may be complemented in the near future by the genetic engineering of antitumor-agent biosynthesis in microorganisms. Biosynthetic gene clusters from several antitumor pathways in actinomycetes are presently being characterized and expressed in order to generate novel drugs. Several novel hydroxylated and glycosylated antitumor-drug derivatives have been produced that show a relaxed substrate specificity for secondary-metabolic enzymes, which opens up the possibility of generating novel drugs by genetic manipulation.

Actinomycetales↗