THE INVIVO CONSTRUCTION OF 4-CHLORO-2-NITROPHENOL ASSIMILATORY BACTERIA
THE INVIVO CONSTRUCTION OF 4-CHLORO-2-NITROPHENOL ASSIMILATORY BACTERIA
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DOI：
10.1007/bf00425158
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发表时间：
1988-06-01
影响因子：
2.8
通讯作者：
KNACKMUSS, HJ
中科院分区：
文献类型：
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作者：
BRUHN, C;BAYLY, RC;KNACKMUSS, HJ
假单胞菌N31是从土壤中分离到的一株分别以3-硝基苯酚和琥珀酸为唯一碳源和氮源的菌株。该菌株表达硝基苯酚加氧酶，并且可以使用2-硝基苯酚或4-氯-2-硝基苯酚作为氮源，消除亚硝酸盐，并分别积累邻苯二酚和3-氯邻苯二酚。儿茶酚没有进一步降解。通过将来自假单胞菌B13或真养产碱杆菌JMP 134（pJP 4）的卤代芳香族降解序列转移到菌株N31，构建了能够利用4-氯-2-硝基苯酚作为唯一碳源和氮源的菌株。使用JMP 134作为供体菌株构建的接合子菌株在3-氯苯甲酸盐上生长，但在2，4-二氯苯氧基乙酸盐上不生长。这是由于2，4-二氯苯氧乙酸单加氧酶和2，4-二氯苯酚羟化酶的非诱导。将质粒从2，4-二氯苯氧乙酸盐阴性转移接合子菌株转移到JMP 134的治愈菌株中，产生也具有相同表型的菌株。这表明在pJP 4中发生了突变以阻止2，4-二氯苯氧乙酸单加氧酶和2，4-二氯苯酚羟化酶的表达。
Pseudomonas sp. N31 was isolated from soil using 3-nitrophenol and succinate as sole source of nitrogen and carbon respectively. The strain expresses a nitrophenol oxygenase and can use either 2-nitrophenol or 4-chloro-2-nitrophenol as a source of nitrogen, eliminating nitrite, and accumulating catechol and 3-chlorocatechol, respectively. The catechols were not degraded further. Strains which are able to utilize 4-chloro-2-nitrophenol as a sole source of carbon and nitrogen were constructed by transfer of the haloaromatic degrading sequences from either Pseudomonas sp. B13 or Alcaligenes eutrophus JMP134 (pJP4) to strain N31. Transconjugant strains constructed using JMP134 as the donor strain grew on 3-chlorobenzoate but not on 2,4-dichlorophenoxyacetate. This was due to the non-induction of 2,4-dichlorophenoxyacetate monooxygenase and 2,4-dichlorophenol hydroxylase. Transfer of the plasmid from the 2,4-dichlorophenoxyacetate negative transconjugant strains to a cured strain of JMP134 resulted in strains which also had the same phenotype. This indicates that a mutation has occurred in pJP4 to prevent the expression of 2,4-dichlorophenoxyacetate monooxygenase and 2,4-dichlorophenol hydroxylase.