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Penzkofer, Alfons ; Atrib, Yama ; Sharmab, Komal ; Veetil, Sindhu Kandoth ; Kateriya, Suneel

Photodynamics of photo-activated BLUF coupled Endonuclease III mutant RmPAE from mesophilic, pigmented bacterium Rubellimicrobium mesophilum strain MSL-20T

Article

Penzkofer, Alfons, Atrib, Yama, Sharmab, Komal, Veetil, Sindhu Kandoth and Kateriya, Suneel (2016) Photodynamics of photo-activated BLUF coupled Endonuclease III mutant RmPAE from mesophilic, pigmented bacterium Rubellimicrobium mesophilum strain MSL-20T. BAOJ Chemistry 2 (2:008), pp. 1-24.

DOI to cite this document: 10.5283/epub.33598


Abstract

The pink to light reddish-pigmented bacterium Rubellimicrobium mesophilum strain MSL-20T contains a BLUF coupled endonuclease III of unknown function. A purified recombinant triple mutated sample of the BLUF coupled endonuclease III (F5Y, N27H, A87W) named RmPAE (Rubellimicrobium mesophilum Photo-Activated Endonuclease) was produced and characterized by optical spectroscopic methods. The BLUF ...

The pink to light reddish-pigmented bacterium Rubellimicrobium mesophilum strain MSL-20T contains a BLUF coupled endonuclease III of unknown function. A purified recombinant triple mutated sample of the BLUF coupled endonuclease III (F5Y, N27H, A87W) named RmPAE (Rubellimicrobium mesophilum Photo-Activated Endonuclease) was produced and characterized by optical spectroscopic methods. The BLUF domain photo-cycling dynamics occurred with high efficient blue-light induced signaling state formation (quantum yield of signaling state formation φs ≈ 0.6), small spectral red-shift (δλs,r ≈ 5.4 nm), and slow thermal activated dark recovery to the receptor state (τrec ≈ 20 min at room temperature). An apparent RmPAE melting temperature of ϑm ≈ 63 °C was determined by stepwise sample heating and absorption spectrum analysis. The photo-degradation of RmPAE in the signaling state was determined by prolonged intense blue-light exposure. An irreversible flavin photo-degradation occurred with quantum yield of φD ≈ 2.6×10-5. Schemes of the photo-cycling and the photo-degradation dynamics are presented. Engineered RmPAE may find application as light guided DNA cutter in optogenetic applications.



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Details

Item typeArticle
Journal or Publication TitleBAOJ Chemistry
PublisherBIO ACCENT
Open Access TypeGold (with APC)
Volume2
Number of Issue or Book Chapter2:008
Page Rangepp. 1-24
Date25 January 2016
Date of publication13 May 2016 09:38
InstitutionsPhysics > Institute of Experimental and Applied Physics > Alumni or Retired Professors > Group Alfons Penzkofer
Identification Number
ValueType
Article ID: 008Other
Dewey Decimal Classification500 Science > 530 Physics
StatusPublished
RefereedYes, this version has been refereed
Created at the University of RegensburgYes
URN of the UB Regensburgurn:nbn:de:bvb:355-epub-335987
Item ID33598

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