Department of Biology, University of Konstanz, Konstanz, Germany.
J Bacteriol. 2013 Jun;195(12):2921-30. doi: 10.1128/JB.00307-13. Epub 2013 Apr 19.
Hypotaurine (HT; 2-aminoethane-sulfinate) is known to be utilized by bacteria as a sole source of carbon, nitrogen, and energy for growth, as is taurine (2-aminoethane-sulfonate); however, the corresponding HT degradation pathway has remained undefined. Genome-sequenced Paracoccus denitrificans PD1222 utilized HT (and taurine) quantitatively for heterotrophic growth and released the HT sulfur as sulfite (and sulfate) and HT nitrogen as ammonium. Enzyme assays with cell extracts suggested that an HT-inducible HT:pyruvate aminotransferase (Hpa) catalyzes the deamination of HT in an initial reaction step. Partial purification of the Hpa activity and peptide fingerprinting-mass spectrometry (PF-MS) identified the Hpa candidate gene; it encoded an archetypal taurine:pyruvate aminotransferase (Tpa). The same gene product was identified via differential PAGE and PF-MS, as was the gene of a strongly HT-inducible aldehyde dehydrogenase (Adh). Both genes were overexpressed in Escherichia coli. The overexpressed, purified Hpa/Tpa showed HT:pyruvate-aminotransferase activity. Alanine, acetaldehyde, and sulfite were identified as the reaction products but not sulfinoacetaldehyde; the reaction of Hpa/Tpa with taurine yielded sulfoacetaldehyde, which is stable. The overexpressed, purified Adh oxidized the acetaldehyde generated during the Hpa reaction to acetate in an NAD(+)-dependent reaction. Based on these results, the following degradation pathway for HT in strain PD1222 can be depicted. The identified aminotransferase converts HT to sulfinoacetaldehyde, which desulfinates spontaneously to acetaldehyde and sulfite; the inducible aldehyde dehydrogenase oxidizes acetaldehyde to yield acetate, which is metabolized, and sulfite, which is excreted.
次牛磺酸(HT;2-氨基乙磺酸)是一种已知的细菌利用物,可作为细菌生长的唯一碳源、氮源和能源;牛磺酸(2-氨基乙磺酸)也是如此;然而,相应的 HT 降解途径仍然未知。基因组测序的脱氮副球菌 PD1222 定量利用 HT(和牛磺酸)进行异养生长,并将 HT 硫释放为亚硫酸盐(和硫酸盐)和 HT 氮作为铵。细胞提取物的酶测定表明,HT 诱导的 HT:丙酮酸转氨酶(Hpa)在初始反应步骤中催化 HT 的脱氨。Hpa 活性的部分纯化和肽指纹图谱-质谱(PF-MS)鉴定了 Hpa 候选基因;它编码了典型的牛磺酸:丙酮酸转氨酶(Tpa)。通过差异 PAGE 和 PF-MS 也鉴定了相同的基因产物,以及强烈诱导的醛脱氢酶(Adh)的基因。这两个基因在大肠杆菌中均过表达。过表达、纯化的 Hpa/Tpa 显示 HT:丙酮酸转氨酶活性。鉴定出的反应产物有丙氨酸、乙醛和亚硫酸盐,但没有亚磺酰乙醛;Hpa/Tpa 与牛磺酸的反应生成稳定的磺酰乙醛。过表达、纯化的 Adh 在 NAD(+) 依赖的反应中,将 Hpa 反应中生成的乙醛氧化为醋酸盐。基于这些结果,可以描绘出 PD1222 菌株中 HT 的以下降解途径。鉴定出的转氨酶将 HT 转化为亚磺酰乙醛,后者自发脱磺化为乙醛和亚硫酸盐;诱导型醛脱氢酶将乙醛氧化为醋酸盐,然后代谢醋酸盐,将亚硫酸盐排泄出去。