Swetha Vandana P, Phale Prashant S
Biotechnology Group, School of Biosciences and Bioengineering, Indian Institute of Technology, Bombay, Powai, Mumbai 400 076, India.
Appl Environ Microbiol. 2005 Oct;71(10):5951-6. doi: 10.1128/AEM.71.10.5951-5956.2005.
Pseudomonas sp. strains C4, C5, and C6 utilize carbaryl as the sole source of carbon and energy. Identification of 1-naphthol, salicylate, and gentisate in the spent media; whole-cell O2 uptake on 1-naphthol, 1,2-dihydroxynaphthalene, salicylaldehyde, salicylate, and gentisate; and detection of key enzymes, viz, carbaryl hydrolase, 1-naphthol hydroxylase, 1,2-dihydroxynaphthalene dioxygenase, and gentisate dioxygenase, in the cell extract suggest that carbaryl is metabolized via 1-naphthol, 1,2-dihydroxynaphthalene, and gentisate. Here, we demonstrate 1-naphthol hydroxylase and 1,2-dihydroxynaphthalene dioxygenase activities in the cell extracts of carbaryl-grown cells. 1-Naphthol hydroxylase is present in the membrane-free cytosolic fraction, requires NAD(P)H and flavin adenine dinucleotide, and has optimum activity in the pH range 7.5 to 8.0. Carbaryl-degrading enzymes are inducible, and maximum induction was observed with carbaryl. Based on these results, the proposed metabolic pathway is carbaryl --> 1-naphthol --> 1,2-dihydroxynaphthalene --> salicylaldehyde --> salicylate --> gentisate --> maleylpyruvate.
假单胞菌属菌株C4、C5和C6利用西维因作为唯一的碳源和能源。在消耗的培养基中鉴定出1-萘酚、水杨酸盐和龙胆酸盐;全细胞对1-萘酚、1,2-二羟基萘、水杨醛、水杨酸盐和龙胆酸盐的氧气摄取;以及在细胞提取物中检测到关键酶,即西维因水解酶、1-萘酚羟化酶、1,2-二羟基萘双加氧酶和龙胆酸盐双加氧酶,这表明西维因是通过1-萘酚、1,2-二羟基萘和龙胆酸盐进行代谢的。在此,我们证明了在以西维因培养的细胞的提取物中存在1-萘酚羟化酶和1,2-二羟基萘双加氧酶活性。1-萘酚羟化酶存在于无膜的胞质部分,需要NAD(P)H和黄素腺嘌呤二核苷酸,并且在pH值7.5至8.0的范围内具有最佳活性。西维因降解酶是可诱导的,以西维因诱导时观察到最大诱导效果。基于这些结果,提出的代谢途径是西维因→1-萘酚→1,2-二羟基萘→水杨醛→水杨酸盐→龙胆酸盐→马来酰丙酮酸。