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Effect of insertional mutations in the pueA and pueB genes encoding two polyurethanases in Pseudomonas chlororaphis contained within a gene cluster
Author(s) -
Howard G.T.,
Mackie R.I.,
Cann I.K.O.,
OheneAdjei S.,
Aboudehen K.S.,
Duos B.G.,
Childers G.W.
Publication year - 2007
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/j.1365-2672.2007.03447.x
Subject(s) - mutant , polyurethane , gene knockout , wild type , gene , chemistry , biology , genetics , microbiology and biotechnology , biochemistry , organic chemistry
Aims: To better understand the role of PueA and PueB from Pseudomonas chlororaphis in polyurethane degradation, the present study was conducted to create insertional mutants in their respective genes. Methods and Results: Growth kinetic studies showed that the pue A knockout mutant had a greater effect than the pue B knockout mutant. The pue A mutant had an 80% decrease in cell density from that of the wild type, while the pue B mutant had an 18% decrease in cell density. Polyurethane utilization followed Michaelis‐Menten kinetics. The pue A and pue B mutants exhibited a 17% and 10% decrease respectively in growth rate using polyurethane when compared with the wild type. Conclusions: In this present study, pue A and pue B, are shown to be part of an ABC transporter gene cluster that consists of seven open reading frames. Mutational analysis results suggest that PueA may play a more major role in polyurethane degradation than PueB based on cell density and growth rates. Significance and Impact of the Study: The results from this study provide a starting point for the eventual enhancement and bioremediation of polyurethane waste. Understanding the role of polyurethane‐degrading enzymes is useful for the creation of strains for this purpose.