A Fluorescence Perspective on the Differential Interaction of Riboflavin and Flavin Adenine Dinucleotide with Cucurbit[7]uril

2010 ◽  
Vol 114 (33) ◽  
pp. 10717-10727 ◽  
Author(s):  
Sharmistha Dutta Choudhury ◽  
Jyotirmayee Mohanty ◽  
Achikanath C. Bhasikuttan ◽  
Haridas Pal
Keyword(s):  
Flavin Adenine Dinucleotide ◽  
Differential Interaction

scholarly journals Flavin Adenine Dinucleotide-Dependent Halogenase XanH and Engineering of Multifunctional Fusion Halogenases

2020 ◽  
Vol 86 (18) ◽  
Author(s):  
Lingxin Kong ◽  
Qing Wang ◽  
Zixin Deng ◽  
Delin You
Keyword(s):  
Protein Engineering ◽  
Fusion Protein ◽  
Natural Product ◽  
Flavin Adenine Dinucleotide ◽  
High Efficiency ◽  
Genetic Manipulation ◽  
Good Alternative ◽  
Limited Information ◽  
Flavin Reductase ◽  
Free Standing

ABSTRACT Xantholipin (compound 1), a polycyclic xanthone antibiotic, exhibited strong antibacterial activities and showed potent cytotoxicity. The biosynthetic gene cluster of compound 1 has been identified in our previous work, and the construction of xanthone nucleus has been well demonstrated. However, limited information of the halogenation involved in compound 1 biosynthesis is available. In this study, based on the genetic manipulation and biochemical assay, we characterized XanH as an indispensable flavin adenine dinucleotide (FAD)-dependent halogenase (FDH) for the biosynthesis of compound 1. XanH was found to be a bifunctional protein capable of flavin reduction and chlorination and exclusively used the NADH. However, the reduced flavin could not be fully and effectively utilized, and the presence of an extra flavin reductase (FDR) and chemical-reducing agent could promote the halogenation. XanH accepted its natural free-standing substrate with angular fused polycyclic aromatic systems. Meanwhile, it exhibited moderate halogenation activity and possessed high substrate specificity. The requirement of extra FDR for higher halogenation activity is tedious for future engineering. To facilitate efforts in engineering XanH derivative proteins, we constructed the self-sufficient FDR-XanH fusion proteins. The fusion protein E1 with comparable activities to that of XanH could be used as a good alternative for future protein engineering. Taken together, these findings reported here not only improve the understanding of polycyclic xanthones biosynthesis but also expand the substrate scope of FDH and pave the way for future engineering of biocatalysts for new active substance synthesis. IMPORTANCE Halogenation is important in medicinal chemistry and plays an essential role in the biosynthesis of active secondary metabolites. Halogenases have evolved to catalyze reactions with high efficiency and selectivity, and engineering efforts have been made to engage the selective reactivity in natural product biosynthesis. The enzymatic halogenations are an environmentally friendly approach with high regio- and stereoselectivity, which make it a potential complement to organic synthesis. FDHs constitute one of the most extensively elucidated class of halogenases; however, the inventory awaits to be expanded for biotechnology applications and for the generation of halogenated natural product analogues. In this study, XanH was found to reduce flavin and halogenated the freely diffusing natural substrate with an angular fused hexacyclic scaffold, findings which were different from those for the exclusively studied FDHs. Moreover, the FDR-XanH fusion protein E1 with comparable reactivity to that of XanH serves as a successful example of genetic fusions and sets an important stage for future protein engineering.

scholarly journals REVERSIBLE ENZYMATIC SYNTHESIS OF FLAVIN-ADENINE DINUCLEOTIDE

1950 ◽  
Vol 182 (2) ◽  
pp. 795-803 ◽  
Author(s):  
Anthony W. Schrecker ◽  
Arthur. Kornberg
Keyword(s):  
Enzymatic Synthesis ◽  
Flavin Adenine Dinucleotide

scholarly journals Differential interaction with TREM2 modulates microglial uptake of modified Aβ species

Glia ◽  
2021 ◽  
Vol 69 (12) ◽  
pp. 2917-2932
Author(s):  
Pranav Joshi ◽  
Florian Riffel ◽  
Kanayo Satoh ◽  
Masahiro Enomoto ◽  
Seema Qamar ◽  
...  
Keyword(s):  
Differential Interaction
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