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  • 1
    Electronic Resource
    Electronic Resource
    Production and biochemical characterization of an α-amylase from the moderate halophile Halomonas meridiana (2000)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 183 (2000), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Extracellular amylase production by the moderate halophile Halomonas meridiana was optimized and the enzyme was characterized biochemically. The highest amylase production was achieved by growing H. meridiana cultures in media with 5% salts and starch, in the absence of glucose until the end of the exponential phase. The amylase exhibited maximal activity at pH 7.0, being relatively stable in alkaline conditions. Optimal temperature and salinity for activity were 37°C and 10% NaCl, respectively. Moreover, activity at salinity as high as 30% salts was detected. Maltose and maltotriose were the main end products of starch hydrolysis, indicating an α-amylase activity.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.2000.tb08935.x
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  • 2
    Electronic Resource
    Electronic Resource
    Expression in Escherichia coli of a cloned β-glucanase gene from Bacillus amyloliquefaciens (1985)
    Springer
    Applied microbiology and biotechnology 22 (1985), S. 63-71 
    Details
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The recombinant phage λG1 has been identified by screening 700 plaques of a Charon 4A library, containing DNA of Bacillus amyloliquefaciens, for phage clones directing the hydrolysis of lichenan in Escherichia coli, as indicated by haloes surrounding plaques on lichenan agar. The gene coding for an endo-β-1.3–1.4-glucanase was recloned within a 3.6 kb EcoRI fragment into the EcoRI site of plasmid pBR322, in both orientations. The location and extent of the bgl gene on the 3.6 kb Bacillus DNA insert was estimated by insertion mutagenesis with transposon Tn5 and restriction mapping of Tn5 insertions within or near to the bgl gene. The β-glucanase synthesized by E. coli harbouring plasmids pEG1 or pEG2 was shown to accumulate mainly in the periplasmic space but β-glucanase activities were also detected extracellulary and in the cytoplasm. The molecular weight of the enzyme synthesized in E. coli harbouring pEG1 was estimated by SDS-polyacrylamide gel electrophoresis to be about 24000. It was shown that the level of bgl gene expression in E. coli varies about 10-fold, depending on the orientation of the 3.6 kb DNA-fragment cloned within the EcoRI site of pBR322. After insertion of HindIII-DNA fragments from phage into the HindIII site of the β-glucanase-high-expression plasmid pEG1, we obtained clones also showing an approximately 10-fold reduction in β-glucanase activites. It was thus concluded that on plasmid pEG1 the leftward acting Apr (PI) promotor of plasmid pBR322 strongly increases the expression in E. coli of the cloned B. amyloliquefaciens bgl gene.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00252158
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  • 3
    Electronic Resource
    Electronic Resource
    DNA repair in Proteus mirabilis (1979)
    Springer
    Molecular genetics and genomics 176 (1979), S. 265-273 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The expression of plasmid R46-mediated recovery and mutagenic function (s) was studied in P. mirabilis, which is normally either weakly or nonmutable after UV exposure. The plasmid was found to confer on P. mirabilis enhanced UV resistance as well as UV-induced mutability for various types of forward mutations and reversion of the thr273 mutation. The plasmid enhanced survival of UV-irradiated phages in P. mirabilis both in unirradiated host cells and with increased efficiency after UV-exposure of host cells, as is characteristic of UV-inducible phage reactivation. Spontaneous mutability of P. mirabilis harboring R46 was about 2 to 7 times higher than that of cells without plasmid, depending on the marker, repair type, and plating density of the cells used. All of these R46-mediated rescue and mutagenic functions require the rec672+ gene function. It is assumed that the plasmid R46 adds functions to P. mirabilis comparable to those deficient in umuC and uvm mutants of E. coli (Kato and Shinoura, 1977; Steinborn, 1978) and that P. mirabilis possesses functions homologous to those controlled in E. coli by the recA + and lexA + genes. The significance of plasmid-mediated rescue and mutagenic functions for bacteria which lack the misrepair branch of mutagenesis, is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00273221
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  • 4
    Electronic Resource
    Electronic Resource
    Functional substitution of the recE gene of Bacillus subtilis by the recA gene of Proteus mirabilis (1984)
    Springer
    Molecular genetics and genomics 195 (1984), S. 516-522 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Rec mutants of Bacillus subtilis have been tested for complementation by the recA gene of Proteus mirabilis (recApm) which was introduced into B. subtilis via the plasmid pHP334. In the recE4 mutant of B. subtilis the plasmid pHP334 restored significantly the defects in RecE functions tested: UV-sensitivity, homologous recombination (transduction and transformation) and prophage induction. Although serological methods to detect the presence of RecApm protein in B. subtilis have been unsuccessful, our results strongly indicate that the recE function of B. subtilis is analogous to the recA function of P. mirabilis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00341456
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  • 5
    Electronic Resource
    Electronic Resource
    A T7 promoter-specific, inducible protein expression system forBacillus subtilis (1996)
    Springer
    Molecular genetics and genomics 250 (1996), S. 230-236 
    Details
    ISSN: 1617-4623
    Keywords: Bacillus subtilis ; T7 RNA polymerase ; Promoter specificity ; Gene expression ; Protein secretion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The adaptation and application of theEscherichia coli T7 RNA polymerase system for regulated and promoter-specific gene expression inBacillus subtilis is reported. The expression cassette used inBacillus subtilis was tightly regulated and T7 RNA polymerase (T7 RNAP) appeared 30 min after induction. The efficiency of T7 promoter-specific gene expression inB. subtilis was studied using one secretory and two cytosolic proteins of heterologous origin. The accumulation ofE. coli β-galactosidase, as well as a 1,4-β-glucosidase fromThermoanaerobacter brockii inB. subtilis after T7 RNAP induction was strongly enhanced by rifampicin inhibition of host RNAP activity. Theα-amylase ofThermoactinomyces vulgaris, a secretory protein, was found to accumulate in the culture supernatant up to levels of about 70 mg/l 10–20 h after T7 RNAP induction, but was also deposited in cellular fractions. The addition of rifampicin inhibitedα-amylase secretion, but unexpectedly, after a short period, also prevented its further (intra)cellular accumulation
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02174183
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  • 6
    Electronic Resource
    Electronic Resource
    DNA repair in Proteus mirabilis (1977)
    Springer
    Molecular genetics and genomics 154 (1977), S. 35-41 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Post-irradiation DNA degradation in P. mirabilis rec + strains after UV irradiation is found to be more extensive in starvation buffer than in growth medium. In growth medium restriction of protein synthesis, but not DNA synthesis, largely prevents the expression of “breakdown limitation”. By the addition of chloramphenicol during post-irradiation incubation in growth medium the expression of break-down limitation was followed and found to occur 20 to 40 min after UV irradiation. Pre-irradiation by a low dose of UV leads after a corresponding time of post-irradiation incubation to breakdown limitation even in starvation buffer after a second UV exposure. Post-irradiation DNA degradation is presumed to be initiated at the sites of DNA lesions which arise at replication points damaged by UV. While pre-starvation restricts the efficiency of postirradiation DNA degradation by the reduction of the number of replication points active at the time of irradiation, caffeine as well as 2,4-dinitrophenol inhibit DNA degradation even in rec - cells probably by the interference with nicking or exonucleoltytic events initiated at those sites in the absence of breakdown limitation. Breakdown limitation is postulated to be due to inducible derepression of REC-functions which lead to the protection and, probably, repair of DNA lesions arising at the replication points following UV exposure.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00265574
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  • 7
    Electronic Resource
    Electronic Resource
    Mathematische Auswertung von Dichtegradienten zur Molekulargewichtsbestimmung von DNS mit Hilfe der EDV 1 (1977)
    Springer
    Genetic resources and crop evolution 25 (1977), S. 267-277 
    Details
    ISSN: 1573-5109
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Description / Table of Contents: Summary A computer programm for the desk-top calculator HP 9810A to analyse sedimentation profiles and to calculate the weight average (Mw) and the number-average molecular weight (Mn) of DNA as well as of their breakage parameters (Nw andNn) is presented. The calculations are based on the radioactivity profiles of tritium-labelled DNA sedimented in alkaline sucrose gradients using a liquid scintillation spectrometer. With the aim of the approximation formula fromBurgi undHershey (1963) the exponentK has been estimated from the sedimentation of standard DNA with standard conditions and yields to 0.353. The data are printed out by mean of a computer-controlled typewriter or as a plot by mean of a computer-controlledX- Y plotter. The drawing program is adapted for different modes of standardisation. The most significant capabilities of the program are the estimation of the molecular weight and breakage number of any fraction or region of the sedimentation profile. In addition the graphic plot permits by use of different standardisation the estimation of quantitative changes under the experimental conditions.
    Abstract: Abstract Описывается програм ма эффективной обраб отки опытных данных для ЭВМHP 9810A. Вычисляются характ еристики (Mw, Mn и дополнит ельно Nw, Nn) для седиментации ДНК в (щелочном) градиенте сахарозы. Предпосылкой этому являются, с одно й стороны, профили радиоактивности, пол ученные при помощи сцинтилляцио нного счётчика, с друг ой —нормирование градиента с помощью стандартных ДНК на основе приближ енной формулы бурги и хершей (1963). В качестве по казателя К в программ е взята полученная в нашей нормировке вел ичина 0,353. Вывод данных и резуль татов осуществляетс я на автоматическом печа тающем устройстве. Кроме тог о предусмотрено граф ическое изображение с помощь ю автоматического рис ующего устройства. Гр афическая часть программы позволяет сопостави ть в количественном о тношении варианты опыта с разл ичной нормировкой, что треб уется для эксперимен тов по репарации ДНК.
    Notes: Zusammenfassung Es wird ein EDV-Programm für den HP 9810 A zur rationellen Auswertung des Sedimentationsverhaltens von DNS im alkalischen Saccharosegradienten, insbesondere zur Berechnung wichtiger Kennzahlen für das Molekulargewicht und für die Bruchhäufigkeiten von DNS beschrieben. Voraussetzung dafür sind Radioaktivitätsprofile und Eichung des Gradienten mit Hilfe von Standard-DNS ausgehend von der Approximationsformel vonBurgi undHershey (1963). Im Programm hat der ExponentK den in unseren Eichläufen ermittelten Wert 0,353. Die Ausgabe der Daten und der Ergebnisse erfolgt als Ausdruck über eine rechnergesteuerte Schreibmaschine. Außerdem ist die graphische Wiedergabe über den rechnergesteuertenx- y-Schreiber vorgesehen. Der graphische Teil des Programms erlaubt es, die Versuchsvarianten (Gradienten) in quantitativer Relation mit unterschiedlicher Normierung auszuwerten, wie es speziell in Experimenten zur DNS-Reparatur notwendig ist.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02014812
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  • 8
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    Antimicrobial Activities and Matrix-Assisted Laser Desorption / Ionization Mass Spectrometry of Bacillus Isolates from the Marine Sponge Aplysina aerophoba (2003)
    Springer
    In:  Marine Biotechnology, 5 (5). pp. 424-434.
    Details
    Publication Date: 2015-08-27
    Description: The aim of this study was to isolate bacteria that are resistant to the strong antimicrobial metabolites characteristic of Aplysina aerophoba. For this purpose, bacterial isolation was performed on agar plates to which sponge tissue extract had been added. Following screening for antifungal and antimicrobial activities, 5 strains were chosen for more detailed analyses. 16S ribosomal DNA sequencing revealed that all isolates belonged to the genus Bacillus, specifically B. subtilis and B. pumilus. Using a combination of matrix-assisted laser desorption/ ionization mass spectrometry typing of whole cells and antimicrobial bioassays against selected reference strains, the bioactive metabolites were identified as lipopeptides.
    Type: Article , PeerReviewed
    Format: text
    DOI: 10.1007/s10126-002-0088-8
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    OceanRep: Article in a Scientific Journal - peer-reviewed
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