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Metabolic shift in the production of corrinoid compounds by Lactobacillus coryniformis in the absence of purines.
Biochimie ( IF 3.3 ) Pub Date : 2019-11-09 , DOI: 10.1016/j.biochi.2019.11.003
Andrea Carolina Torres 1 , Mariano Elean 1 , Elvira María Hebert 1 , Lucila Saavedra 1 , María Pía Taranto 1
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Lactobacillus coryniformis CRL 1001 and L. reuteri CRL 1098 have the complete genes necessary to synthesize pseudo-cobalamin as final product in a vitamin B12 free commercial medium. Unlike vitaminB12 (the most biologically active form), the pseudo-cobalamin contains adenine instead of 5,6-dimethlbenzimidazole (DMB) in the Coα-ligand. Considering the vitamin B12-gene clusters of these bacteria, the aim of this work was to analyze the production of corrinoids with DMB (vitamin B12) instead of adenine (pseudo-B12) as lower ligand base in a vitamin B12 free chemically defined medium (CDM) without purines. Genome-wide screening of genes related to purine metabolism showed that both strains possess all pur genes necessary for the synthesis of inositol monophosphate, the main precursor for purine biosynthesis. Accordingly, both strains were able to grow in B12 free CDM without purines, with the supplementation of different synthetic intermediaries. Isolated compounds with positive vitamin B12 activity were quantified and characterized by LC/MS-MS. Total corrinoids values were higher for both strains in comparison to those obtained in vitaminB12 free commercial medium. Interestingly, CRL 1001 strain synthesized cobalamin, suggesting that this strain is able to activate DMB as nitrogenous base instead adenine when it is in excess in a purine-free medium. The present paper represents the first demonstration of a partial metabolic shift to produce vitamin B12 in a Lactobacillus strain.

中文翻译:

在不存在嘌呤的情况下,谷氨酸乳杆菌产生的类corrinoid化合物发生代谢转移。

变形乳酸杆菌CRL 1001和罗伊氏乳杆菌CRL 1098具有在不含维生素B12的商业培养基中合成拟钴胺素作为最终产物所必需的完整基因。与维生素B12(最具生物活性的形式)不同,假钴胺素在Coα配体中包含腺嘌呤而不是5,6-二甲基苯并咪唑(DMB)。考虑到这些细菌的维生素B12基因簇,这项工作的目的是在不含维生素B12的化学成分确定的培养基中,以DMB(维生素B12)代替腺嘌呤(伪B12)作为较低的配体基础来分析类胡萝卜素的产生( CDM)不含嘌呤。对嘌呤代谢相关基因进行全基因组筛选显示,这两个菌株均具有合成肌醇一磷酸(嘌呤生物合成的主要前体)所需的所有pur基因。因此,两种菌株都能够在无嘌呤的情况下在不含B12的CDM中生长,并补充了不同的合成中间体。定量分离的具有正维生素B12活性的化合物,并通过LC / MS-MS进行了表征。与在无维生素B12的商业培养基中获得的菌株相比,两种菌株的总类固醇含量值都更高。有趣的是,CRL 1001菌株合成了钴胺素,表明该菌株在无嘌呤的培养基中过量时能够激活DMB作为含氮碱而不是腺嘌呤。本论文代表了在乳酸杆菌菌株中产生部分维生素B12的新陈代谢转变的第一个证明。定量分离的具有正维生素B12活性的化合物,并通过LC / MS-MS进行了表征。与在无维生素B12的商业培养基中获得的菌株相比,两种菌株的总类固醇含量值都更高。有趣的是,CRL 1001菌株合成了钴胺素,表明该菌株在不含嘌呤的培养基中过量时,能够将DMB活化为含氮碱而不是腺嘌呤。本论文代表了乳酸菌菌株中部分代谢转移产生维生素B12的第一个证明。定量分离的具有正维生素B12活性的化合物,并通过LC / MS-MS进行了表征。与在无维生素B12的商业培养基中获得的菌株相比,两种菌株的总类固醇含量值都更高。有趣的是,CRL 1001菌株合成了钴胺素,表明该菌株在不含嘌呤的培养基中过量时,能够将DMB活化为含氮碱而不是腺嘌呤。本论文代表了乳酸菌菌株中部分代谢转移产生维生素B12的第一个证明。这表明当该菌株在无嘌呤的培养基中过量时,能够将DMB活化为含氮碱而不是腺嘌呤。本论文代表了乳酸菌菌株中部分代谢转移产生维生素B12的第一个证明。这表明当该菌株在无嘌呤的培养基中过量时,能够将DMB活化为含氮碱而不是腺嘌呤。本论文代表了在乳酸杆菌菌株中产生部分维生素B12的新陈代谢转变的第一个证明。
更新日期:2019-11-11
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