Abstract:
:Optochin-resistant mutant and wild-type diaphorases were purified approximately 300-fold by a combination of batch adsorption and column chromatography with diethylaminoethyl cellulose, and were characterized with regard to their pH optima, sensitivity to optochin inhibition and heat inactivation, Michaelis constants with flavine mononucleotide (FMN) and reduced nicotinamide adenine dinucleotide (NADH), and inhibition constants with optochin hydrochloride. The pH optima of the purified diaphorases were similar, but the purified diaphorases from the optochin-resistant strains were approximately four to five times more resistant to heat inactivation at 45 C than was the wild-type diaphorase. Purified diaphorase preparations from the optochin-resistant pneumococci had greater activities per milligram of protein and were more resistant to optochin inhibition than the preparation from the optochin-sensitive pneumococcus. Michaelis constants for FMN and NADH were similar; however, the inhibition constants of the optochin-resistant diaphorases were four to eight times greater than that of the optochin-sensitive diaphorase. Optochin hydrochloride produced a noncompetitive type of inhibition with FMN as substrate but a competitive type of inhibition with NADH as substrate. Optochin hydrochloride produced an approximately 10-fold increase in the Michaelis constant for NADH. The concentration of drug required to produce this effect was, however, greater with the mutant diaphorases than with the wild-type diaphorase. Optochin hydrochloride quenched the fluorescence of riboflavine. This phenomenon did not appear to be related to the diaphorase-inhibitory activity of the drug, however, since the pH requirements of the two reactions were different. Quenching of riboflavine fluorescence by optochin hydrochloride increased with a rise in pH, whereas inhibition of diaphorase activity by optochin hydrochloride was greater at pH 6.8 than at pH 7.6.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Kreger AS,Olsen RHdoi
10.1128/JB.96.4.1029-1036.1968subject
Has Abstractpub_date
1968-10-01 00:00:00pages
1029-36issue
4eissn
0021-9193issn
1098-5530journal_volume
96pub_type
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doi:10.1128/JB.107.2.499-504.1971
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pub_type: 杂志文章
doi:10.1128/JB.117.3.1002-1009.1974
更新日期:1974-03-01 00:00:00
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更新日期:1992-01-01 00:00:00
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pub_type: 杂志文章
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更新日期:1989-05-01 00:00:00
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更新日期:1972-03-01 00:00:00
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更新日期:1972-01-01 00:00:00
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更新日期:1996-08-01 00:00:00
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更新日期:1984-08-01 00:00:00
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更新日期:2011-11-01 00:00:00
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更新日期:2004-02-01 00:00:00
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更新日期:1999-10-01 00:00:00
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更新日期:1976-04-01 00:00:00
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更新日期:2003-08-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.85.5.1094-1103.1963
更新日期:1963-05-01 00:00:00
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更新日期:2016-04-14 00:00:00