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对介导芳香族β-葡萄糖苷代谢的菊欧文氏菌arb基因的分析。

Analysis of the Erwinia chrysanthemi arb genes, which mediate metabolism of aromatic beta-glucosides.

作者信息

el Hassouni M, Chippaux M, Barras F

机构信息

Laboratoire de Chimie Bactérienne, Centre National de la Recherche Scientifique, Marseille, France.

出版信息

J Bacteriol. 1990 Nov;172(11):6261-7. doi: 10.1128/jb.172.11.6261-6267.1990.

Abstract

Erwinia chrysanthemi is one of the few members of the family Enterobacteriaceae that is capable of metabolizing most of the naturally occurring beta-glucosides. We previously isolated the clb genes, which allow the use of the disaccharide cellobiose as well as the aromatic beta-glucosides arbutin and salicin. We report here the isolation of the arb genes, which permit fermentation of the aromatic beta-glucosides only. Establishment of a functional Arb system in Escherichia coli depended on the presence of the phosphotransferase system and on the activation by the cyclic AMP-cyclic AMP receptor protein complex. Strains carrying mini-Mu-induced LacZ fusions to the arb genes were used to analyze arb genes organization and function. Three arb genes (arbG, arbF, and arbB) were identified and organized in this order. Genetic and structural evidence allowed us to assign a phospho-beta-glucosidase and a permease activity to the ArbB and ArbF proteins, respectively. Several Lac+ arb-lacZ insertions were introduced into the E. chrysanthemi chromosome. Both ArbG- and ArbF- strains were unable to ferment the aromatic beta-glucosides, whereas ArbB- strains were impaired only in salicin fermentation. None of the mutations in the arb genes affected cellobiose metabolism. The expression of the arb genes was substrate inducible and required the ArbF permease and, possibly, the ArbG protein. Collectively, our results underline the resemblance between the naturally expressed E. chrysanthemi arbGFB and the cryptic E. coli bglGFB operons, yet the arbG gene product seemed unable to activate E. coli bgl operon expression.

摘要

菊欧文氏菌是肠杆菌科中少数能够代谢大多数天然存在的β-葡萄糖苷的成员之一。我们之前分离出了clb基因,该基因使得细胞能够利用二糖纤维二糖以及芳香族β-葡萄糖苷熊果苷和水杨苷。我们在此报告arb基因的分离情况,该基因仅允许芳香族β-葡萄糖苷的发酵。在大肠杆菌中建立功能性的Arb系统取决于磷酸转移酶系统的存在以及环腺苷酸-环腺苷酸受体蛋白复合物的激活。携带mini-Mu诱导的与arb基因融合的LacZ的菌株用于分析arb基因的组织和功能。鉴定出了三个arb基因(arbG、arbF和arbB),并按此顺序排列。遗传和结构证据使我们能够分别将磷酸-β-葡萄糖苷酶和通透酶活性赋予ArbB和ArbF蛋白。将几个Lac + arb-lacZ插入片段导入菊欧文氏菌染色体。ArbG和ArbF缺陷型菌株均无法发酵芳香族β-葡萄糖苷,而ArbB缺陷型菌株仅在水杨苷发酵方面受损。arb基因中的任何突变均不影响纤维二糖代谢。arb基因的表达是底物诱导型的,并且需要ArbF通透酶,可能还需要ArbG蛋白。总的来说,我们的结果强调了天然表达的菊欧文氏菌arbGFB与隐秘的大肠杆菌bglGFB操纵子之间的相似性,然而,argG基因产物似乎无法激活大肠杆菌bgl操纵子的表达。

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