A novel formamidase is required for riboflavin biosynthesis in invasive bacteria.
A novel formamidase is required for riboflavin biosynthesis in invasive bacteria.
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DOI：
10.1016/j.jbc.2022.102377
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发表时间：
2022-09
影响因子：
4.8
通讯作者：
Roje, Sanja
中科院分区：
文献类型：
--
作者：
Yurgel, Svetlana N.;Johnson, Skylar A.;Rice, Jennifer;Sa, Na;Bailes, Clayton;Baumgartner, John;Pitzer, Josh E.;Roop, R. Martin I. I. I. I.;Roje, Sanja
核黄素(Rf)是氧化还原辅因子FMN和FAD的前体，其生物合成在细菌中被认为是众所周知的，所有的途径酶都被认为是已知的和必不可少的。我们以前的研究对这一观点提出了挑战，结果表明，在苜蓿中华根瘤菌中，编码催化RF生物合成途径初始步骤的酶的Ribba基因的缺失只会导致黄素分泌减少，而不是RF营养不良。这一发现让我们假设Ribba参与了黄素的生物合成以供分泌，而米氏葡萄球菌有另一种酶为内部细胞新陈代谢执行这一功能。在这里，我们鉴定并生化鉴定了一种新的甲酰胺酶(SMc02977)，该酶参与了米氏链霉菌胞内功能的RF的产生。我们将这种催化剂命名为Sm-BrbF，它从早期的RF前体2-amino-5-formylamino-6-ribosylamino-4(3H)-pyrimidinone 5‘-磷酸中释放甲酸盐，生成2，5-二氨基-6-核糖氨基-4(3H)-嘧啶酮5’-磷酸。我们发现该酶的同源物存在于许多细菌中，在根瘤菌目中含量非常丰富，来自流产布鲁氏菌和青枯病自由杆菌的序列同源物补充了Sm1021ΔSmC02977突变体的RF营养缺陷症。此外，我们还证明了巴氏杆菌酶(Bab20247，BaBrbF)也是一种2-amino-5-formylamino-6-ribosylamino-4(3H)-pyrimidinone 5‘-磷酸甲酰胺酶，并且Bab20247突变株是一种RF营养缺陷体，比野生型菌株具有更低的细胞内感染水平。最后，我们证明了Sm-BrbF和BaBrbF与其他RF生物合成途径的酶直接相互作用。总之，我们的结果为细菌中RF生物合成的错综复杂提供了新的见解。
Biosynthesis of riboflavin (RF), the precursor of the redox cofactors FMN and FAD, was thought to be well understood in bacteria, with all the pathway enzymes presumed to be known and essential. Our previous research has challenged this view by showing that, in the bacterium Sinorhizobium meliloti, deletion of the ribBA gene encoding the enzyme that catalyzes the initial steps on the RF biosynthesis pathway only causes a reduction in flavin secretion rather than RF auxotrophy. This finding led us to hypothesize that RibBA participates in the biosynthesis of flavins destined for secretion, whereas S. meliloti has another enzyme that performs this function for internal cellular metabolism. Here, we identify and biochemically characterize a novel formamidase (SMc02977) involved in the production of RF for intracellular functions in S. meliloti. This catalyst, which we named Sm-BrbF, releases formate from the early RF precursor 2-amino-5-formylamino-6-ribosylamino-4(3H)-pyrimidinone 5′-phosphate to yield 2,5-diamino-6-ribosylamino-4(3H)-pyrimidinone 5′-phosphate. We show that homologs of this enzyme are present in many bacteria, are highly abundant in the Rhizobiales order, and that sequence homologs from Brucella abortus and Liberobacter solanacearum complement the RF auxotrophy of the Sm1021ΔSMc02977 mutant. Furthermore, we show that the B. abortus enzyme (Bab2_0247, Ba-BrbF) is also an 2-amino-5-formylamino-6-ribosylamino-4(3H)-pyrimidinone 5′-phosphate formamidase, and that the bab2_0247 mutant is a RF auxotroph exhibiting a lower level of intracellular infection than the wildtype strain. Finally, we show that Sm-BrbF and Ba-BrbF directly interact with other RF biosynthesis pathway enzymes. Together, our results provide novel insight into the intricacies of RF biosynthesis in bacteria.
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影响因子：
4.8
作者：
Ren, J;Kotaka, M;Stammers, DK
通讯作者：
Stammers, DK
影响因子：
3.7
作者：
Ruy Bonomi, Hernan;Ines Marchesini, Maria;Alberto Goldbaum, Fernando
通讯作者：
Alberto Goldbaum, Fernando
DOI：
10.1107/s0907444913011402
发表时间：
2013-09-01
期刊：
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY
影响因子：
--
作者：
Singh, Mirage;Kumar, Pankaj;Karthikeyan, Subramanian
通讯作者：
Karthikeyan, Subramanian
DOI：
10.1073/pnas.96.22.12275
发表时间：
1999-10-26
影响因子：
11.1
作者：
Phillips, DA;Joseph, CM;Volpin, H
通讯作者：
Volpin, H
影响因子：
3.2
作者：
HUANG, HC;HE, SY;COLLMER, A
通讯作者：
COLLMER, A