Ruff Jürgen, Denger Karin, Cook Alasdair M
Department of Biology, University of Konstanz, D-78457 Konstanz, Germany.
Biochem J. 2003 Jan 15;369(Pt 2):275-85. doi: 10.1042/BJ20021455.
The facultatively anaerobic bacterium Alcaligenes defragrans NKNTAU was found to oxidize taurine (2-aminoethanesulphonate) with nitrate as the terminal electron acceptor. Taurine was transaminated to 2-sulphoacetaldehyde. This was not converted into sulphite and acetate by a "sulphoacetaldehyde sulpho-lyase" (EC 4.4.1.12), but into sulphite and acetyl phosphate, which was identified by three methods. The enzyme, which required the addition of phosphate, thiamin diphosphate and Mg(2+) ions for activity, was renamed sulphoacetaldehyde acetyltransferase (Xsc; EC 2.3.1.-). Inducible Xsc was expressed at high levels, and a three-step 11-fold purification yielded an essentially homogeneous soluble protein, which was a homotetramer in its native form; the molecular mass of the subunit was found to be between about 63 kDa (SDS/PAGE) and 65.3 kDa (matrix-assisted laser-desorption ionization-time-of-flight MS). The N-terminal and two internal amino acid sequences were determined, and PCR primers were generated. The xsc gene was amplified and sequenced; the derived molecular mass of the processed protein was 65.0 kDa. The downstream gene presumably encoded the inducible phosphate acetyltransferase (Pta) found in crude extracts. The desulphonative enzymes ("EC 4.4.1.12") from Achromobacter xylosoxidans NCIMB 10751 and Desulfonispora thiosulfatigenes GKNTAU were shown to be Xscs. We detected at least three subclasses of xsc in Proteobacteria and in Gram-positive bacteria, and they comprised a distinct group within the acetohydroxyacid synthase supergene family. Genome sequencing data revealed xsc genes in Burkholderia fungorum (80% sequence identity) and Sinorhizobium meliloti (61%) with closely linked pta genes. Different patterns of regulation for the transport and dissimilation of taurine were hypothesized for S. meliloti and B. fungorum.
兼性厌氧菌恶臭产碱菌NKNTAU被发现能以硝酸盐作为末端电子受体氧化牛磺酸(2-氨基乙磺酸盐)。牛磺酸经转氨作用生成2-磺基乙醛。2-磺基乙醛并非通过“磺基乙醛磺基裂解酶”(EC 4.4.1.12)转化为亚硫酸盐和乙酸盐,而是转化为亚硫酸盐和乙酰磷酸,这是通过三种方法鉴定出来的。该酶的活性需要添加磷酸盐、硫胺素二磷酸和镁离子,被重新命名为磺基乙醛乙酰转移酶(Xsc;EC 2.3.1.-)。可诱导的Xsc大量表达,经过三步11倍的纯化得到一种基本均一的可溶性蛋白,其天然形式为同四聚体;亚基的分子量在约63 kDa(SDS/PAGE)至65.3 kDa(基质辅助激光解吸电离飞行时间质谱)之间。测定了N端和两个内部氨基酸序列,并生成了PCR引物。扩增并测序了xsc基因;加工后蛋白质的推导分子量为65.0 kDa。下游基因可能编码粗提物中发现的可诱导的磷酸乙酰转移酶(Pta)。木糖氧化无色杆菌NCIMB 10751和硫代硫酸盐脱硫孢菌GKNTAU的脱硫酶(“EC 4.4.1.12”)被证明是Xsc。我们在变形菌门和革兰氏阳性菌中检测到至少三个xsc亚类,它们在乙酰羟酸合酶超基因家族中构成一个独特的群体。基因组测序数据显示,在真菌伯克霍尔德菌(序列同一性80%)和苜蓿中华根瘤菌(61%)中存在xsc基因,且与紧密相连的pta基因相邻。推测苜蓿中华根瘤菌和真菌伯克霍尔德菌对牛磺酸的转运和异化存在不同的调控模式。