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枯草芽孢杆菌乌头酸酶的纯化及其N端氨基酸序列与citB基因序列的相关性。

Purification of aconitase from Bacillus subtilis and correlation of its N-terminal amino acid sequence with the sequence of the citB gene.

作者信息

Dingman D W, Sonenshein A L

出版信息

J Bacteriol. 1987 Jul;169(7):3062-7. doi: 10.1128/jb.169.7.3062-3067.1987.

DOI:10.1128/jb.169.7.3062-3067.1987
PMID:3110133
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC212349/
Abstract

The DNA sequence for the promoter region of the Bacillus subtilis citB gene has been determined. Presumed "-10" and "-35" regions of the promoter have been identified, and transcriptional and translational start points of citB have been located. To correlate the DNA sequence of citB with the amino acid sequence of its presumed product, aconitase, it was necessary to devise a scheme for purification of this labile enzyme. This procedure relies on the ability to restore enzyme activity at each stage of purification by incubation in a reducing buffer containing a source of ferrous ions. B. subtilis aconitase appears to be a monomer with a molecular weight of approximately 120,000. The amino-terminal amino acids of aconitase fit the sequence predicted by analysis of the citB gene. Thus, citB codes for aconitase.

摘要

已确定枯草芽孢杆菌citB基因启动子区域的DNA序列。已鉴定出该启动子假定的“-10”和“-35”区域，并确定了citB的转录和翻译起始点。为了将citB的DNA序列与其假定产物乌头酸酶的氨基酸序列相关联，有必要设计一种纯化这种不稳定酶的方案。该程序依赖于在含有亚铁离子源的还原缓冲液中孵育，从而在纯化的每个阶段恢复酶活性的能力。枯草芽孢杆菌乌头酸酶似乎是一种分子量约为120,000的单体。乌头酸酶的氨基末端氨基酸与通过citB基因分析预测的序列相符。因此，citB编码乌头酸酶。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d802/212349/f55d98e2dca9/jbacter00197-0165-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d802/212349/928da52d722f/jbacter00197-0164-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d802/212349/f55d98e2dca9/jbacter00197-0165-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d802/212349/928da52d722f/jbacter00197-0164-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d802/212349/f55d98e2dca9/jbacter00197-0165-a.jpg

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