从恶臭假单胞菌OU83中纯化2,3-二羟基联苯1,2-双加氧酶及该基因（bphC）的特性分析
Purification of 2,3-dihydroxybiphenyl 1,2-dioxygenase from Pseudomonas putida OU83 and characterization of the gene (bphC).
作者信息
Khan A A, Wang R F, Nawaz M S, Cao W W, Cerniglia C E
机构信息
Microbiology Division, Food and Drug Administration, Jefferson, Arkansas 72079, USA.
出版信息
Appl Environ Microbiol. 1996 May;62(5):1825-30. doi: 10.1128/aem.62.5.1825-1830.1996.
Abstract
The 2,3-dihydroxybiphenyl 1,2-dioxygenase (2,3-DBPD) of Pseudomonas putida OU83 was constitutively expressed and purified to apparent homogeneity. The apparent molecular mass of the native enzyme was 256 kDa, and the subunit molecular mass was 32 kDa. The data suggested that 2,3-DBPD was an octamer of identical subunits. The nucleotide sequence of a DNA fragment containing the bphC region was determined. The deduced protein sequence for 2,3-DBPD consisted of 292 amino acid residues, with a calculated molecular mass of 31.9 kDa, which was in agreement with data for the purified 2,3-DBPD. Nucleotide and amino acid sequence analyses of the bphC gene and its product, respectively, revealed that there was a high degree of homology between the OU83 bphC gene and the bphC genes of Pseudomonas cepacia LB400 and Pseudomonas pseudoalcaligenes KF707.
摘要
恶臭假单胞菌OU83的2,3-二羟基联苯1,2-双加氧酶（2,3-DBPD）组成型表达并纯化至表观均一。天然酶的表观分子量为256 kDa，亚基分子量为32 kDa。数据表明2,3-DBPD是由相同亚基组成的八聚体。测定了包含bphC区域的DNA片段的核苷酸序列。推导的2,3-DBPD蛋白质序列由292个氨基酸残基组成，计算分子量为31.9 kDa，这与纯化的2,3-DBPD的数据一致。对bphC基因及其产物分别进行核苷酸和氨基酸序列分析，结果显示OU83 bphC基因与洋葱伯克霍尔德菌LB400和类产碱假单胞菌KF707的bphC基因之间存在高度同源性。
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