Cyclic analogue of tambjamine YP1 alkaloid isolated from ...Cyclic analogue of tambjamine YP1...
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Cyclic analogue of tambjamine YP1 alkaloid isolated from marine bacterium Pseudoalteromonas citrea Katherine J. Picott a Julie A. Deichert a , Ella M. Dekemp a , Gabriele Schatte a , Françoise Sauriol a , and Avena C. Ross *a a Department of Chemistry, Queen’s University, Kingston, ON, Canada. *Corresponding author: Avena C. Ross, [email protected] Supplementary Information Table S1. Proteins in the tam pathway, their putative functions, and the percent identity between P. tunicata and P. citrea proteins 1–4 . Protein Putative function Identity (%) TamA Adenylation and acyl-carrier protein 70 TamB Peptidyl-carrier protein 69 TamC Rieske oxygenase 83 TamD Aminotransferase 78 TamE L-prolyl-AMP ligase 56 TamF -ketoacyl synthase 49 TamG L-prolyl-PCP dehydrogenase 74 TamH Thioester reductase and aminotransferase 71 TamJ Oxidase/dehydrogenase 53 TamK Outer membrane lipoprotein sorting protein 74 TamL Permease 75 TamM Permease 75 TamN ATP-transporter, ATP-binding protein 78 TamO Hypothetical protein 56 TamP SAM dependent O-methyl transferase 69 TamQ Condensation enzyme 65 TamR No assigned function 66 TamS Phosphopantetheinyl transferase 42 TamT Dehydrogenase 62 Electronic Supplementary Material (ESI) for MedChemComm. This journal is © The Royal Society of Chemistry 2019
Transcript of Cyclic analogue of tambjamine YP1 alkaloid isolated from ...Cyclic analogue of tambjamine YP1...
CyclicanalogueoftambjamineYP1alkaloidisolatedfrommarinebacteriumPseudoalteromonascitrea
KatherineJ.PicottaJulieA.Deicherta,EllaM.Dekempa,GabrieleSchattea,FrançoiseSauriola,andAvenaC.Ross*a
aDepartmentofChemistry,Queen’sUniversity,Kingston,ON,Canada.
*Correspondingauthor:AvenaC.Ross,[email protected]
SupplementaryInformation TableS1.Proteinsinthetampathway,theirputativefunctions,andthepercentidentitybetweenP.tunicataandP.citreaproteins1–4.
Protein Putativefunction Identity(%)
TamA Adenylationandacyl-carrierprotein 70TamB Peptidyl-carrierprotein 69
TamC Rieskeoxygenase 83TamD Aminotransferase 78TamE L-prolyl-AMPligase 56
TamF 𝛽-ketoacylsynthase 49TamG L-prolyl-PCPdehydrogenase 74TamH Thioesterreductaseandaminotransferase 71
TamJ Oxidase/dehydrogenase 53TamK Outermembranelipoproteinsortingprotein 74TamL Permease 75
TamM Permease 75TamN ATP-transporter,ATP-bindingprotein 78TamO Hypotheticalprotein 56
TamP SAMdependentO-methyltransferase 69TamQ Condensationenzyme 65
TamR Noassignedfunction 66TamS Phosphopantetheinyltransferase 42TamT Dehydrogenase 62
Electronic Supplementary Material (ESI) for MedChemComm.This journal is © The Royal Society of Chemistry 2019
TambjamineMYP1structuraldata
Figure S1. 1H NMR spectrum of tambjamineMYP1 collected in CDCl3in a 500MHz field. Solvents and internalstandardtrimethylsilane(TMS)arelabelled.
FigureS2.13CNMRspectrumoftambjamineMYP1collectedinCDCl3ina700MHzfield.
CDCl3
H2O TMS
CDCl3
FigureS3.1H-1HCOSYNMRspectrumoftambjamineMYP1.CollectedinCDCl3ina500MHzfield.
FigureS4. 1H-1HTOCSYNMRspectrumof tambjamineMYP1withunknown impurity,collected inCDCl3 ina500MHzfield.
CDCl3 H2O
FigureS5. 1H-13CHSQCNMRspectrumof tambjamineMYP1withunknown impurity, collected inCDCl3 ina500MHzfield.
TableS2.CrystaldataandstructurerefinementfortambjamineMYP15,6.
CrystalData
Empiricalformula C24H35N3O5
Formulaweight 445.55
CrystalColor,Habit yellow,plate-like
Crystaldimensions(mm) 0.186×0.128×0.090
Crystalsystem triclinic
Spacegroup P1[2]
Unitcellparameters a(Å) 9.8155(5)b(Å) 11.1719(5)c(Å) 11.1821(5)α(°) 91.5571(16)β(°) 95.2218(16)γ(°) 101.2158(17)V(Å3) 1196.53(10)Zb 2F(000) 480Density(ρcalcd) 1.237Mg/m3Absorptioncoefficient(µ) 0.087mm-1
DataCollectionandRefinementConditions
CDCl3 H2O
Diffractometer BrukerAXSD8VentureDuodiffractometerRadiation monochromatedMoKα ( 𝜆 =0.71073Å)Temperature -93(2)°C[180(2)K]Scantype ϕ-andω-scans(1.5º/frame,15sexposure/frame,9
sets)
Thetarangefordatacollection 2.533to26.396°
Completenesstotheta=25.242° 99.9%
Reflectionscollected 38405
Indexranges -12≤h≤12,-13≤k≤13,-13≤l≤13
Independentreflections[Fo2≥-3σ(Fo
2)] 4891[Rint=0.0736]
Observedreflections[Fo2>2σ(Fo
2)] 3437
Absorptioncorrectionmethod multi-scan[SADABS]
AnomalousDispersion Forallnon-hydrogenatoms
Structuresolutionmethod Directmethods(SHELXT-2014)
Refinementmethod Full-matrixleast-squaresonF2
(SHELXL-2017/1)i
FunctionMinimized Σw(|Fo|2-|kFc|
2)2(k:overallscalefactor)
Weighingscheme,w
w=[σ(Fo2)+(aP)2+(bP)]-1
w=[σ(Fo2)+(0.0723P)2+1.2456P]-1
P-factor [Max(Fo2,0)+2Fc
2]/3Data/restraints/parameters 4891[Fo
2≥-3σ(Fo2)]/5/305
Reflection(observed)/parameterratio 11:1Reflection(data)/parameterratio 16:1Goodness-of-fit on F2
GooF = {Σ[w(Fo2-Fc
2)2]/(n - p)}1/2
n: number of reflections, p: number of parameters
1.018
FinalRindices
R1=[Σ||Fo|-|Fc||]/[Σ|Fo|]for[Fo2>2σ(Fo
2)] 0.0641
wR2={[Σw(Fo2-Fc
2)2]/[Σw(Fo2)2]}1/2[alldata] 0.1795
Max.Shift/ErrorinFinalCycle 0.000
Largestdifferencepeakandhole 0.833and-0.603e-/Å3
Transmissionfactor(min) 0.7071[SADABS]Transmissionfactor(max) 0.7454[SADABS]
FigureS6.FullasymmetricunitoftambjamineMYP1crystalstructure,thermalellipsoidsaredrawnto30%probability.
FigureS7.UnitcelloftambjamineMYP1X-raystructure.(A)Viewatoriginofunitcell,(B)viewonabfaceofunitcell.Hydrogenatomsandformicacidmoleculeomittedforclarity.Theedgecoloursrefertothea(blue),b(green),andc(red)coordinates.
(B)(A)
Generalsynthesisinformation
Commercially available reagents were used throughout without further purification. Reactions were
carriedoutunderanargonatmosphereusingoven-driedglassware.1HNMRspectrawererecordedusingaBruker
Avance-400 spectrometeroperatingat400MHz.AnhydrousDCE (Sure/Seal™)wasobtained fromSigma-Aldrich
andusedasreceived.
SynthesisofTambjamineBE-18591·HCl(Z)-N-((4'-methoxy-1H,5'H-[2,2'-bipyrrol]-5'-ylidene)methyl)dodecan-1-aminehydrochloride
TambjamineBE-18591wassynthesizedbymodificationofthemethodofSaggiomoetal7.Toasolutionof
4-methoxy-2,2’-bipyrrole-5-carboxaldehyde(10mg,0.053mmol)anddodecylamine(13mg,0.068mmol)inDCE(1
mL)wasadded2dropsAcOH(cat.).Thereactionmixturewasheatedat60°Cfor3h,thencooledtort,diluted
withCH2Cl2(5mL),andwashedwith1MHCl(3×10mL).TheorganicfractionwasdriedoverMgSO4,subjectedto
filtration,andconcentratedinvacuotodeliverthetitlecompoundasayellowoil(18mg,89%). 1HNMR(400MHz;CDCl3):δ 13.70(brs,1H),10.62(brs,1H),9.52-9.40(m,1H),7.33(d,J=14.9Hz,1H),
7.06(dd,J=1.4and2.7Hz,1H),6.73(dd,J=1.4and3.7Hz,1H),6.28(2.6and3.8Hz),5.94(d,J=2.2Hz,1H),3.92
(s,3H),3.51-3.45(m,2H),1.75(p,J=7.3Hz,2H),1.34-1.25(m,18H),0.87(t,J=6.7Hz,3H).
Physicalandspectraldatawerefoundtobeconsistentwiththosereported7.
NH
NH
HN
OMe
Me Cl
pKadeterminationinformation
FigureS8.AbsorbancespectraatvaryingpHof(A)the lineartambjamineBE-19591(TB=baseform,TBH+=acidform)and(B)thetambjamineMYP1(cTB=baseform,cTBH+=acidform).Spectratakenin0.1MNaClin1:1(v/v)MeCN/H2O,pHadjustedwith0.1-0.01MHCl.TableS3.Theabsorbancemaximaofeachcompoundintheirbasicandacidicforms,andthecalculatedpKa.Compound Base𝝀max(nm) Acid𝝀max(nm) pKaTambjamineBE-18591 368 400 9.9±0.2TambjamineMYP1 385 420 10.4±0.3 The pKa was calculated using Equation S1 from previous literature8, this was done with absorbancemeasurementsatseveralpHvaluesusingboth𝜆max(basicandacidic)forcalculations,thesewerethenaveraged.Inthe equation,A refers to the experimental absorbance at a particularwavelength,Ab- is the absorbanceof thebaseformatthiswavelength,andAHistheabsorbanceoftheacidicform.EquationS1 𝑝𝐾! = 𝑝𝐻 + log (!!!!
!
!"!!)
References:1 C.Burke,T.Thomas,S.EganandS.Kjelleberg,Environ.Microbiol.,2007,9,814–818.2 A.M.Cerdeño,M.J.BibbandG.L.Challis,Chem.Biol.,2001,8,817–829.3 S.F.Altschul,W.Gish,W.Miller,E.W.MyersandD.J.Lipman,J.Mol.Biol.,1990,215,403–410.4 D.M.Withall,S.W.HaynesandG.L.Challis,J.Am.Chem.Soc.,2015,137,7889–7897.5 G.M.Sheldrick,ActaCrystallogr.,2008,A64,112–122.6 G.M.Sheldrick,ActaCrystallogr.,2015,A71,3–8.7 V.Saggiomo,S.Otto,I.Marques,V.Felix,T.TorrobaandR.Quesada,Chem.Commun.,2012,48,5274–
5276.8 G.S.Patterson,J.Chem.Educ.,1999,76,395–398.
0.00
0.50
1.00
300 320 340 360 380 400 420 440 460
AU
Wavelength(nm)
pH12.13pH11.59pH10.76pH9.56pH8.93pH8.66pH8.51
TB
TBH+
0.00
0.10
0.20
320 340 360 380 400 420 440 460 480
AU
Wavelength(nm)
pH11.15pH10.15pH9.55pH9.03pH8.72pH8.36
cTB
cTBH+(A) (B)
