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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 5 (1963), S. 147-165 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A number of factors was found to affect the cup plate assay for lysine with Streptococcus faecalis ATCC 6057. Hydroxylysine, aspartic acid, and Tween 80 were found to be among the essential components of the assay medium. Removal of any of these three components produced diffuse growth zones or otherwise unsatisfactory plates. The inclusion of asparagine and additional dipotassium phosphate to the medium was found to be advantageous but not an absolute requirement. It was important not to autoclave the hydroxylysine and the additional dipotassium phosphate together with the rest of the assay medium, but to add these components to the sterilized medium immediately before pouring the plates. With certain concentrations of hydroxylysine in the medium, the addition of L-glutamine was found to potentiate the effect of hydroxylysine and thereby improve the lysine plates. The effect of graded amounts of the above mentioned medium components and of the pH of the assay medium was also studied. Increasing the sterilization time of the medium from 5 to 10 min. at 120°C. improved the readability of the lysine plates and led to more easily reproducible results. Factors influencing the preparation of the inoculum for the lysine plates were also studied: the age of the stab culture, the amount of inoculum used per plate, and the influence of the nature of the suspension medium used for washing the inoculum.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 5 (1963), S. 131-146 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A sensitive cup plate assay method for lysine on large plates using S. faecalis ATCC 6057 as the test organism is described in detail. Well-defined zones of dense growth were obtained in a rectilinear dose - response range from 5 to 3000 μg. of L-lysine/ml. Comparative assays on twelve different samples were carried out using the turbidimetric tube assay method with Leuc. mesenteroides ATCC 8042 and the cup plate method with S. faecalis ATCC 6057. Good agreement between the two assay methods was obtained for five samples. For the remaining seven samples the tube assay values were appreciably higher than those obtained by the plate method. This discrepancy is discussed. Pure lysine added to various sample extracts could be quantitatively recovered. The specificity of the lysine requirement of S. faecalis ATCC 6057 in the plate assay was also investigated. Under certain experimental conditions, a weak and variable response was obtained with D-lysine and the polyamines cadaverine, spermine, and spermidine. There was generally no difficulty in distinguishing the D-lysine and polyamine growth zones from the L-lysine growth zones. This growth effect of D-lysine and polyamines could be diminished and even completely abolished by increasing the DL-hydroxylysine hydrochloride level in the plate medium. The lysine peptides glycyl-L-lysine and L-lysylglycine gave excellent growth zones of the same appearance as those produced by L-lysine. The activity of these two peptides in cup tests were on a molar basis about 70% of that of L-lysine.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    Library Location Call Number Volume/Issue/Year Availability
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