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  • 1
    Electronic Resource
    Electronic Resource
    Copenhagen : International Union of Crystallography (IUCr)
    Acta crystallographica 50 (1994), S. 225-227 
    ISSN: 1399-0047
    Source: Crystallography Journals Online : IUCR Backfile Archive 1948-2001
    Topics: Chemistry and Pharmacology , Geosciences , Physics
    Notes: Lipase from Chromobacterium viscosum has been purified to homogeneity and crystallized in a form suitable for X-ray diffraction analysis from 10-14% polyethylene glycol 4000 and 10-14% 2-methyl-2,4-pentane diol at pH 6.4 in the presence of 0.25%(w/v) n-octyl-β-D-glucopyranoside. These crystals belong to space group P21212 with refined lattice constants a = 41.1 Å, b= 156.8, c = 43.6 Å, indicating a cell content of one monomer per asymmetric unit of the crystal. The crystals diffract to a resolution of 2.2 Å.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Clinica Chimica Acta 75 (1977), S. 313-319 
    ISSN: 0009-8981
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Medicine
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 799 (1996), S. 0 
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    World journal of microbiology and biotechnology 9 (1993), S. 243-247 
    ISSN: 1573-0972
    Keywords: Bacterial immobilized cells ; liquid-drying ; preservation ; microbial biosensors ; xenobiotics detection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A simple and effective method for the drying of immobilized bacterial cells to be used directly in a microbial biosensor for measurement of activity is reported. As a case example, plasmid-bearing cells of Alcaligenes eutrophus JMP 134, DSM 4058 were immobilized on various carriers and liquid-dried. The dried cell-matrix was used directly after rehydration/reactivation as the biological component of a biosensor for determining the concentration of xenobiotic compounds in the environment. Good viability results were obtained after long-term storage and cells exhibited no loss of plasmids responsible for the 2,4-dichlorophenoxyacetic acid (2,4-D) degradation. The activity of the cells for 2,4-D was proved using a respiration electrode. No time-consuming, repeated cell cultivation and harvesting was required, as the cells preserved from a single batch served as a continuous source for activity measurements. Many other microbial cultures can be preserved by this method and the cells preserved in the form of immobilized dried cell-matrix can be used directly to perform enzymatic tests, complex biochemical conversions and for production in the reactors. The dried cell-matrix can serve as a stable interchangeable component for a multipurpose biosensor.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Fresenius' journal of analytical chemistry 366 (2000), S. 769-779 
    ISSN: 1432-1130
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract In parallel to the continuous development of increasingly more sophisticated physical and chemical analytical technologies for the detection of environmental pollutants, there is a progressively more urgent need also for bioassays which report not only on the presence of a chemical but also on its bioavailability and its biological effects. As a partial fulfillment of that need, there has been a rapid development of biosensors based on genetically engineered bacteria. Such microorganisms typically combine a promoter-operator, which acts as the sensing element, with reporter gene(s) coding for easily detectable proteins. These sensors have the ability to detect global parameters such as stress conditions, toxicity or DNA-damaging agents as well as specific organic and inorganic compounds. The systems described in this review, designed to detect different groups of target chemicals, vary greatly in their detection limits, specificity, response times and more. These variations reflect on their potential applicability which, for most of the constructs described, is presently rather limited. Nevertheless, present trends promise that additional improvements will make microbial biosensors an important tool for future environmental analysis.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Fresenius' journal of analytical chemistry 364 (1999), S. 584-589 
    ISSN: 1432-1130
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract A sensitive dipstick assay for 2,4-dichlorophenoxyacetic acid (2,4-D) detection was developed. The assay was based on the competitive reaction of 2,4-D and enzyme tracer with monoclonal antibodies immobilised on an Ultrabind® membrane. The binding of enzyme tracer on the test strip was determined by a simple, portable reflectometer as remission at 657 nm. Using this technique, 2,4-D could be detected in a concentration range of 0.5 μg/L to 100 μg/L. The center point of the 2,4-D test was found at a concentration of 6 μg/L. Cross-reactivity with 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) as determined by this dipstick assay was 2.5% and 3% by standard ELISA technique using microtiter plates. The assay was applied in the detection of 2,4-D in real water samples, and sensitivity was comparable to spiked water samples. If combined with an effective extraction procedure that results in recovery rates of 90%, the dipstick assay can be used to monitor human exposure to 2,4-D from contamination in water, from oranges and in testing urine samples.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Applied microbiology and biotechnology 45 (1996), S. 600-606 
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract  Geotrichum candidum was found to produce a lactate oxidase. The enzyme was purified by gel filtration and ion-exchange chromatography. The purified lactate oxidase showed a molecular mass of 50 kDa under denaturing and about 400 kDa under non-denaturing conditions. Transmission electron micro-scopy analysis confirmed an octameric structure. FMN was found to be a cofactor for this enzyme. Polarographic studies confirmed an oxygen uptake by the lactate oxidase. The enzyme showed specificity towards the L isomer of lactate and did not oxidise pyruvate, fumarate, succinate, maleate and ascorbate. It was stable at alkaline pH and also for 15 min at 45°C. The addition of glycerol and dextran 500 000 to the enzyme sample enhanced storage stability.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Applied microbiology and biotechnology 49 (1998), S. 405-410 
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract An efficient expression system for the previously only weakly expressed thermophilic lipase BTL2 (Bacillus thermocatenulatus lipase 2) was developed for the production of large amounts of lipase in Escherichia coli. Therefore, the gene was subcloned in the pCYTEXP1 (pT1) expression vector downstream of the temperature-inducible λ promoter PL. Three different expression vectors were constructed: (i) pT1-BTL2 containing the mature lipase gene, (ii) pT1-preBTL2 containing the prelipase gene and (iii) pT1-OmpABTL2 containing the mature lipase gene fused to the signal peptide of the OmpA protein, the major outer membrane protein of E. coli. With pT1-BTL2 and pT1-preBTL2, comparable expression levels of 7000–9000 U/g cells were obtained independently of the E. coli host. In contrast, with E. coli JM105 harbouring pT1-OmpABTL2, 660 000 soluble lipase U/g cells was produced, whereas, with E. coli DH5α and BL321, production levels of 30 000 U/g cells were achieved. However, most of the lipase remained insoluble but active after cell breakage because of the unprocessed OmpA signal peptide. A simple cholate extraction followed by proteinase K cleavage and ultrafiltration allowed the isolation of 1.15 × 106 units of 90% pure mature lipase/wet cells.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A disposable-type microbial sensor was developed for the determination of both the biochemical oxygen demand for nitrification (N-BOD) and inhibiting effects on nitrifying bacteria. The sensor was based on the respiratory activity of nitrifying bacteria immobilized on a miniature oxygen electrode. Typical response times for measuring N-BOD of ammonium standard solutions as well as of wastewater samples were in the range of 6–12 min. A dynamic evaluation of the signals after a measuring time of 120 s also resulted in good reproducibility and sensitivity. A daily profile of a municipal sewage plant was recorded, comparing the biosensor data with two standard methods. For the measurement of nitrification-inhibiting effects a 120-s dynamic signal evaluation was preferred to a steady-state method because of the long recovery times resulting from extended exposure to inhibitors. However, steady-state measurement techniques allowed allylthiourea detection with a ten times higher sensitivity. Because of the advantages of this miniaturized electrode, e.g. short response time, simple measuring procedure and low costs of production, this sensor system is considered to be suitable for commercial application in environmental analysis.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Naturwissenschaften 81 (1994), S. 323-325 
    ISSN: 1432-1904
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Natural Sciences in General
    Type of Medium: Electronic Resource
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