This paper focuses on the research of the bioconversion of 1,3-propanediol by Klebsiella pneumoniae. The linear correlation of cell growth and 1,3-propanediol synthesis was found. An equation of the relationship betwe...
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This paper focuses on the research of the bioconversion of 1,3-propanediol by Klebsiella pneumoniae. The linear correlation of cell growth and 1,3-propanediol synthesis was found. An equation of the relationship between cell growth and biocatalysis was *** the analysis of metabolism, it was discovered that the cell regulated the NADH production by cell growth in order to supply enough reductive equivalent for enzyme catalysis. A conclusion was drawn that the cell growth was coupled with the reactivation of a key-enzyme which catalyzes 1,3-propanediol production in Klebsiella pneumoniae.
Catechol 2,3-dioxygenase(C23O)is the key enzyme of aromatic substance degradation by Pseudomonas sp..In order to establish a simple assay of C23O activity during the whole-cell catalysis of Pseudomonas putida mt-2,C23...
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Catechol 2,3-dioxygenase(C23O)is the key enzyme of aromatic substance degradation by Pseudomonas sp..In order to establish a simple assay of C23O activity during the whole-cell catalysis of Pseudomonas putida mt-2,C23O was induced by utilizing sodium benzoate acid as the sole carbon source,and its activity was determined in whole cells by the amended protocol of pure enzyme *** suspending the cells with potassium phosphate buffer,the substrate was added and the accumulation of 2-hydroxymuconic semialdehyde was measured by a UV757CRT spectrophotometer at 375 *** activity of C23O was evaluated by the climbing slope of time course curve of the UV *** this means,the Km for catechol and C23O in whole cells was 34.67 μmol·L-1,while Vmax was 0.29 μmol·min-1·(mg dry cell)-1,both of which differed from those for pure enzyme by 2—3 orders of *** eliminate the cell wall barrier for substrate permeation,a cationic surfactant,n-dodecyltrimethylammonium bromide,was used to pre-treat the *** 0.1 g·L-1 dodecyl trimethyl ammonium bromide(DTAB) treated for 30 min,the maximum C23O activity could be achieved,which was consistent with the result of treated cells by beads *** the present study,a feasible and simple method was put forward for the apparent enzyme activity assay intracells which could be conveniently applied to the whole-cell biocatalysis or to environmental bioremediation.
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