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Cloning and Expression of Vitreoscilla Hemoglobin Gene in Burkholderia sp. Strain DNT for Enhancement of 2,4‐Dinitrotoluene Degradation
Author(s) -
Patel Sangeeta M.,
Stark Benjamin C.,
Hwang KwangWoo,
Dikshit Kanak L.,
Webster Dale A.
Publication year - 2000
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp9901421
Subject(s) - strain (injury) , burkholderia , pseudomonas , microbiology and biotechnology , chemistry , cloning (programming) , biochemistry , yeast extract , recombinant dna , biology , gene expression , gene , bacteria , fermentation , genetics , anatomy , computer science , programming language
The gene ( vgb ) encoding the hemoglobin (VHb) of Vitreoscilla sp. was cloned into a broad host range vector and stably transformed into Burkholderia (formerly Pseudomonas ) sp. strain DNT, which is able to degrade and metabolize 2,4‐dinitrotoluene (DNT). Vgb was stably maintained and expressed in functional form in this recombinant strain (YV1). When growth of YV1, in both tryptic soy broth and minimal salts broth containing DNT and yeast extract, was compared with that of the untransformed strain, YV1 grew significantly better on a cell mass basis ( A 600 ) and reached slightly higher maximum viable cell numbers. YV1 also had roughly twice the respiration as strain DNT on a cell mass basis, and in DNT‐containing medium, YV1 degraded DNT faster than the untransformed strain. YV1 cells pregrown in medium containing DNT plus succinate showed the fastest degradation: 100% of the initial 200 ppm DNT was removed from the medium within 3 days.