TY - JOUR
T1 - Isolation and Characterization of Tn5-Induced Mutants of Pseudomonas paucimobilis UT26 Defective in γ-Hexachlorocyclohexane Dehydrochlorinase (LinA)
AU - Nagata, Yuji
AU - Imai, Ryozo
AU - Sakai, Akira
AU - Fukuda, Masao
AU - Yano, Keiji
AU - Takagi, Masamichi
N1 - Funding Information:
Acknowledgments. We thank H. Wada and K. Senoo for their advice and Chie Matsumoto for critical reading of the manuscript. This work was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, and Culture of Japan.
PY - 1993
Y1 - 1993
N2 - Pseudomonas paucimobilis UT26 grows on γ-hexachlorocyclohexane (γ-HCH) as a sole source of carbon and energy. Tn5 mutation was introduced into UT26, and two kinds of mutants defective in γ-HCH degradation were phenotypically isolated; one (UT64) completely lacked the activity to degrade γ-HCH, while the other (UT61) retained a very low level of activity. Tagging and sequencing analysis showed that both mutants had a Tn5 insertion at the same site of the linA (γ-HCH dehydrochlorinase encoding) gene. However, UT61 had an additional rearrangement, which could be the cause of its retaining a low level of activity. An in vitro complementation test with a crude extract from UT64 plus partially purified LinA protein showed that LinA was essential not only for the first-step reaction (γ-HCH to γ-pentachlorocyclohexene; γ-PCCH), but also for the second-step reaction (γ-PCCH to compound B) of γ-HCH degradation in UT26.
AB - Pseudomonas paucimobilis UT26 grows on γ-hexachlorocyclohexane (γ-HCH) as a sole source of carbon and energy. Tn5 mutation was introduced into UT26, and two kinds of mutants defective in γ-HCH degradation were phenotypically isolated; one (UT64) completely lacked the activity to degrade γ-HCH, while the other (UT61) retained a very low level of activity. Tagging and sequencing analysis showed that both mutants had a Tn5 insertion at the same site of the linA (γ-HCH dehydrochlorinase encoding) gene. However, UT61 had an additional rearrangement, which could be the cause of its retaining a low level of activity. An in vitro complementation test with a crude extract from UT64 plus partially purified LinA protein showed that LinA was essential not only for the first-step reaction (γ-HCH to γ-pentachlorocyclohexene; γ-PCCH), but also for the second-step reaction (γ-PCCH to compound B) of γ-HCH degradation in UT26.
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U2 - 10.1271/bbb.57.703
DO - 10.1271/bbb.57.703
M3 - Article
C2 - 7686793
AN - SCOPUS:0027598830
SN - 0916-8451
VL - 57
SP - 703
EP - 709
JO - Bioscience, Biotechnology and Biochemistry
JF - Bioscience, Biotechnology and Biochemistry
IS - 5
ER -