Spektrophotometrische Methode zur Bestimmung von Penicillinase

Rossmann, Irene; Siegmund, Peter; Körber, Friedrich

doi:10.1007/bf00546385

Cited by 1 publication

(4 citation statements)

References 7 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The aim of our study was to estimate F and K m of a Bacillus cereus 0-lactamase using a spectrophotometric method (15) with enzymatically hydrolysable penicillins as substrates, and to examine the influence of several inhibitors on the interaction of enzyme and substrates. shown to consist chiefly of a type I 0-lactamase with penicillinase activity, accompanied by a very small proportion of a type II 0-lactamase with cephalosporinase activity.…”

Section: Kinetische Untersuchungen Und Hemmungsstudien An Einer ß-Lacmentioning

confidence: 99%

“…The catalytic activity of the |3-lactamase was determined as described by Rossmann et al (15). The reaction of p^chlormercuribenzoate and penicilloic acid obtained from enzymatic hydrolysis of the examined penicillin was monitored spectrophoto metrically at ë = 250 nm.…”

Section: Kinetic Propertiesmentioning

confidence: 99%

“…Again measurements were carried put as described byRossmann et al (15). Again measurements were carried put as described byRossmann et al (15).…”

Section: Enzyme Inhibitionmentioning

confidence: 99%

“…Absorption coefficients of the examined penicillins were calculated as described by Rossmann et al (15). The corresponding penicilloic acid was obtained by enzymatic hydrolysis.…”

Section: Absorption Coefficientsmentioning

confidence: 99%

See 3 more Smart Citations

Kinetic and Inhibition Studies of Bacillus cereus β-Lactamase Using a Spectrophotometric Method

Münch¹,

Wombacher²,

Körber³

1981

CCLM

Self Cite

View full text Add to dashboard Cite

The use of a Spectrophotometric method is reported for the characterization of a /Mactamase (EC 3.5.2.6) from Bacillus cereus. Absorption coefficients of the mercaptids of various penicillins were determined with this method. The enzyme was kinetically characterized using penicillins.Inhibition studies with Bacillus cereus 0-lactamase and various penicillins showed a substrate type of inhibition, indicating an additional binding site for substrates without catalytic activity.The dissociation constant of this binding site was determined and the influence of this binding site upon the catalytic activity is discussed. Studies with /J-lactamase-stable penicillins as inhibitors and various penicillins showed different types of inhibition, which indicated the presence of an additional catalytically inactive binding site. Experiments with clavulanic acid, a /Mactamase inhibitor without remarkable intrinsic antibacterial activity, showed a mixed type of inhibition. Based on the hypothesis for the existence of more than one substrate binding site on the enzyme, clavulanic acid was found to be bonded to both the catalytic active and the catalytic inactive binding site. Kinetische Untersuchungen und Hemmungsstudien an einer ß-Lactamase aus Bacillus cereus mit einer spektrophoto· metrischen MethodeZusammenfassung: Wir haben eine photometrische Methode zur Bestimmung von Penicillinen für die Charakterisierung einer ß-Lactamase (EC 3.5.2.6) ms Bacillus cereus angewendet und die molaren Absorptionskoeffizienten der aus verschiedenen Penicillinen gebildeten Mercaptide bestimmt. Das kinetische Verhalten des Enzyms wurde unter Verwendung verschiedener Penicilline überprüft. Untersuchungen mit hohen Substratkqnzentrationen ergaben für die ß-Lactamase aus Bacillus cereus mit verschiedenen spaltbaren Penicillinen das Bild einer Substratüberschußhemmung. Aufgrund dieses Hemmtyps wird neben der katalytisch aktiven Stelle eine weitere, substratbindende Stelle auf der untersuchten 0-Lactamase vermutet. Die Dissoziationskonstänte dieser Bindungsstelle wird bestimmt und der Einfluß dieser nicht katalytisch aktiven Stelle auf die katalytisch aktive Stelle diskutiert. Versuche mit jS-Lactamase-festen Penicillinen als Inhibitor und verschiedenen Penicillinen ergaben unterschiedliche Hemmtypen, die unter Einbeziehung der postulierten katalytisch inaktiven substratbindenden Stelle erklärt werden. Clavulansäure, ein ß-Lactamäse-Inhibitor ohne nennenswerte eigene antimik-robieUe Aktivität, zeigte das Bild eines gemischten Hemmtyps. Unter Zugrundelegung der Mehrbindungsstellenhypothese besetzt Clavulansäure sowohl die katalytisch aktive, als auch die katalytisch inaktive Bindungsstelle. , ible penicillin from enzymatic inactivation. The inhibi-In u on tion of a Bacillus cereus 0-lactamase by oxacillin, Since the discovery of peniciUinase in 1940 by Abraham methici y in ^d nafcillin has been described as com-& Chain (1) and the recognition of the importance of pe titive (6, 7), but other observations indicate that a 0-lactamases in the resistance o...

show abstract

Section: Kinetische Untersuchungen Und Hemmungsstudien An Einer ß-Lacmentioning

confidence: 99%

Section: Kinetic Propertiesmentioning

confidence: 99%

“…Again measurements were carried put as described byRossmann et al (15). Again measurements were carried put as described byRossmann et al (15).…”

Section: Enzyme Inhibitionmentioning

confidence: 99%

“…Absorption coefficients of the examined penicillins were calculated as described by Rossmann et al (15). The corresponding penicilloic acid was obtained by enzymatic hydrolysis.…”

Section: Absorption Coefficientsmentioning

confidence: 99%

See 2 more Smart Citations