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德氏乳杆菌保加利亚亚种胱硫醚β-裂合酶编码基因patC的分离及菌株间酶生物合成变异性的分子分析

Isolation of the patC gene encoding the cystathionine beta-lyase of Lactobacillus delbrueckii subsp. bulgaricus and molecular analysis of inter-strain variability in enzyme biosynthesis.

作者信息

Aubel Dominique, Germond Jacques Edouard, Gilbert Christophe, Atlan Danièle

机构信息

Unité de Microbiologie et Génétique, UMR 5122, Université Claude Bernard-Lyon 1, bât. Lwoff, 10 rue Dubois, F-69622 Villeurbanne Cedex, France1.

Nestlé Research Centre, Nestec Ltd, Vers-chez-les-Blancs, CH-1000 Lausanne 26, Switzerland2.

出版信息

Microbiology (Reading). 2002 Jul;148(Pt 7):2029-2036. doi: 10.1099/00221287-148-7-2029.

Abstract

The patC gene encoding the cystathionine beta-lyase (CBL) of Lactobacillus delbrueckii subsp. bulgaricus NCDO 1489 was cloned and expressed in Escherichia coli. Overexpression of CBL complemented the methionine auxotrophy of an E. coli metC mutant, demonstrating in vivo that this enzyme functions as a CBL. However, PatC is distinguishable from the MetC CBLs by a low identity in amino acid sequence, a sensitivity to iodoacetic acid, greater thermostability and a lower substrate affinity. Homologues of patC were detected in the 13 Lb. delbrueckii strains studied, but only seven of them showed CBL activity. In constrast to CBL(+) strains, all CBL-deficient strains analysed were auxotrophic for methionine. This supports the hypothesis that CBLs from lactobacilli are probably involved in methionine biosynthesis. Moreover, the results of this study suggest that post-transcriptional mechanisms account for the differences in CBL activities observed between strains of Lb. delbrueckii.

摘要

编码德氏乳杆菌保加利亚亚种NCDO 1489胱硫醚β-裂解酶(CBL)的patC基因在大肠杆菌中被克隆并表达。CBL的过表达弥补了大肠杆菌metC突变体的甲硫氨酸营养缺陷,在体内证明了这种酶作为CBL发挥作用。然而,PatC与MetC CBLs不同,其氨基酸序列同一性较低,对碘乙酸敏感,热稳定性更高且底物亲和力更低。在所研究的13株德氏乳杆菌菌株中检测到了patC的同源物,但其中只有7株表现出CBL活性。与CBL(+)菌株相比,所有分析的CBL缺陷菌株对甲硫氨酸均为营养缺陷型。这支持了乳杆菌中的CBL可能参与甲硫氨酸生物合成的假说。此外,本研究结果表明转录后机制是造成德氏乳杆菌菌株间观察到的CBL活性差异的原因。

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