Supplementary Materials - MDPI

1 downloads 15 Views 3MB Size Report
В. сепосерасіа. B. cepacia. B. contaminas. B. diffusa. B. dolosa. B. lata. B. latens. B. metallica. B. multivorans. B. pseudomultivorans. B. pyrrocinia. B. seminalis.

Mar. Drugs 2016, 14, 83

S1 of S18

Supplementary Materials: Antimicrobial Activity of Monoramnholipids Produced by Bacterial Strains Isolated from Ross Sea (Antarctica) Pietro Tedesco, Isabel Maida, Fortunato Palma Esposito, Emiliana Tortorella, Karolina Subko, Chidinma Christiana Ezeofor, Ying Zhang, Jioji Tabudravu, Marcel Jaspars, Renato Fani and Donatella de Pascale Table of contents: Figure S1. Phylogenetic trees (using the complete deletion option) of the Arthrobacter (A), Pseudomonas (B) and Psychrobater (C) genera. Figure S2. 1H-NMR spectrum of compound 1 in CD3OD at 600 MHz. Figure S3. Edited-HSQC NMR spectrum of compound 1 in CD3OD. Figure S4. COSY NMR spectrum of compound 1 in CD3OD. Figure S5. HMBC spectrum of compound 1 in CD3OD at 600 MHz. Figure S6. 1H-NMR spectrum of compound 1 in DMSO-d6 at 400 MHz. Figure S7. 1H-NMR spectrum of compound 2 in CD3OD at 600 MHz. Figure S8. Edited-HSQC NMR spectrum of compound 2 in CD3OD. Figure S9. COSY NMR spectrum of compound 2 in CD3OD. Figure S10. HMBC NMR spectrum of compound 2 in CD3OD. Figure S11. 1H-NMR spectrum of compound 2 in DMSO-d6 at 400 MHz. Figure S12. 2D ROESY spectrum of compound 2 in DMSO-d6 with Rhamnose protons labelled. Figure S13. Compound 2 showing selected ROE correlations of the rhamnose moiety. Figure S14. 1H-NMR spectrum of compound 3 in CD3OD at 600 MHz. Figure S15. Edited-HSQC spectrum of compound 3 in CD3OD at 600 MHz. Figure S16. COSY NMR spectrum of compound 3 in CD3OD at 600 MHz. Figure S17. HMBC NMR spectrum of compound 3 in CD3OD. Figure S18. HRESIMS spectrum (M + Na)+ of compound 1. Figure S19. MS/MS data of compound 1. Figure S20. HRESIMS spectrum (M + Na)+ of compound 2. Figure S21. MS/MS data of compound 2. Figure S22. HRESIMS spectrum (M + Na)+ of compound 3. Figure S23. MS/MS data for compound 3. Table S1. Cross- streaking experiments of BTN isolates against a wide panel of Bcc strains. The Bcc strains highlighted in red correspond to type strains. Symbols: +, growth; ±, reduced growth; -, no growth; C+, positive controls, i.e., Bcc strains grown in the absence of the tester strain(s). Abbreviations: Ps, Pseudomonas; Ar, Arthrobacter; CF, Cystic Fibrosis; Env; environmental.

Mar. Drugs 2016, 14, 83

S2 of S18

Arthrobacter _hist

s_X80 738 ns_X83406

cter Art X 807 _m hro 45 yso ba ren Ar cte s_A t hr r _ J61 so l ob Ar 7 i _E ac 482 th ter F6 ro 60 _b Ar ba 74 erg th cte 8 ro e rei r_ ba _A ar ct dle J6 er 09 ye _c 63 n re sis 0 at _A in J5 ol 51 yt ic 16 us 3 _D 88 21 1

hor m iae_

80739

9628

nae_X

itensis_AJ60

rob a

Arth

89

Arth

roba cter_ p

rotop

nicotia bacter_

0

10

81

84

15

96

40

2267

s (T); A-6; AF10

hlorophenolicu Arthrobacter_c

hro

Arthrobacter_phenanthrenivorans_AM176541

_p

ter

ac

Art

Ar t hr ob

98 19

AJ 31 50 ari 09 et i 69 ba s_ 75 cte A Arth 25 r _t J6 73 e rob 39 cti_ act 83 AJ er _ 49 0 63 flav 9 us_ 82 Arth 9 AB roba 537 6859 cter_ 168 agilis _X 8 Arthro 11 0 bacter_ 748 1 niigate 62 8 nsis_A B 2485 26 Arthrobacter 21 _defluvii_AM 409361

e_

_D Q

us

ne

_t um ba

te r

te rra

ac

ub

ro b

te r

_s

Ar th

ac ro b Ar th

Arthrobacte

(A)

Arthro

88 45 94

73

Arth

Arth

Art

roba

rob a

r_livingstonen sis

bacter_

cter_

cter

_alp

stacke b

p syc

_GQ406811

randtii

hrola

ctop

_AJ64

hilus

0198

82 _A F inus bac 1341 _GQ ter BT 79 Ar _ps 227 t N4 ych 413 Ar hrob A roc rth th a h c r ro itin ob ter ba i ph ac _r t er ilus cte am _A r_ os B5 pa us 886 _X sc 33 80 en 74 s_ 2 X8 07 40

71

hro

89 98 27 90 98 AB 27 e_ AB za 36 ry l a_ 07 _o ico X8 er m 091 is_ ct hu 235 rm F ba r_ ifo ro s_A cte lob th ran ba r _g Ar ivo 2 ro cte th lfon 069 72 r a A F 33 _su ob ter e_A t hr 58 ma Ar bac 0741 lero hro s_X8 _ sc e r Art n e acte 77 5 m og rob 5 lychro Arth r _po bacte 8 X8340 Arthro xydans_

te

11

er_o 94 hrobact 2 Art

ac

r_su bacte 1709 _AM93 Arthro tarcticus 82 ter_an rthrobac 60 A 50 ssicus_AB0719 Arthrobacter_ru

44

97 1

b ro

2

Arthrobacter_sanguinis_EU086805

54 44 afaciens_X807 Arthrobacter_ure 6 405 ens_X83 sc re 04 98 acter_au 5125 Arthrob J A 6 _ 5 licus 07 78 ajaco 834 rogu 7 r_nit is_X 852 _ilic bacte r 2 4 ro h te 2 B Art 21 1 90 bac A 0 o _ r 35 ilus Arth F2 liph A 0 a 7 s_ alk 88 hu 53 ter_ 27 r op 33 bac l ot AJ hro X9 s_ thy _ Art e u is se r _m ns _ro cte we ba ter ac ol u hro t b r w A r_ hro

th Ar

81

60

Arthrobacter_nicotinovorans_X80743 76

t Ar

06

5 63 Y1 67 LI 3 ); 61 bi _ (T AJ m _ s s o an rh licu ler r_ no 2 he ot o cte 606 rop cry ba h r o c J60 r sy _A cte th sis _p r n ba Ar e le o t 061 ac 606 t hr gue ob _A J Ar ker t hr nsis er _ t ie Ar c tr o ba ang hro 9 r _g 66 Art 8340 acte us_X rob 39 lfure Arth

30 29

poiete

idinolovora

51

ko

Q4 ;G

5 88

27

ter _

32

ac

hro

51

s_ um AJ mi 24 ns 34 i i ree _X 21 bac ns 93 ter is_ Arth 35 _lu A rob 4 Y1 t eo acte 1 lu 1 64 r _g s_A and 96 00 J24 ave 342 Arth nsis 92 roba 2 _A J cter_ 316 57 citre 140 us_X Arthro 87 bacter_ 8073 crysta 7 llo Art

0.02

31

_c

lbu

Arthro

_a

31

on _m

te r

Arthrobacter _arila

r te

ter

46 39

r te ac

ac ac

ac

_ ter ac

b ro

b ro ro b

4 236 54 725 J29 X 80 93 _A 7 oli_ cae 82 a_c 39 pho igell 6 J asi /Sh A 6 n ti_ ichia er _ 82 70 n her e act 39 b 50 Esc igm J6 hro p 31 Art _A i l l AJ te ti_ as en _c um ob t hr Ar

th Ar th Ar Ar t hr ob

Ar t hr ob

Ar th

Arthrobacter_uratoxydans_X83410

Supplementary material

Mar. Drugs 2016, 14, 83

Pseudo

67 Pseudomomonas_viridiflava_AY180972 nas_congela 33 ns_AJ49282 33 Pseudom 8 24 PPseudom onas_syringa se on e_DQ31 5 8866 P udom as_ficu

21 28 22 65

6621 domonas_lut ea_AY364 537 Pseudomonas_gramini s_Y11150

do

Ps

(B)

sere

on

AJ u s 1 a 0 s PsPseeu eud domona _ars ndel 3599 49282 0 7 eu uddomom ona s_k enic ii_AF 6 do om o ona s_ ilon oxy 058 m o nas s_ thiv ens dan 286 on na _ br er is_ s_ as s_ bra ass val AJ FN6 4 _f co ss ica ens 292 re rru ica ce is 4 5213 de g ce a _A 26 rik ata ar rum F10 sb _D um _A 03 er 84 _E F 1 23 ge 0 U 0 ns 12 39 032 13 1 is 88 _A J2 49 38 2 80

do m P s on eu as_ do m mo Ps on dom cre Ps nas_ eud as_ on mo eu x a r do caen omo ant s_p ico Ps mona i_E nas hom ara lora U 6 _m a f u t a eu s Ps dom _pse 206 end rina lva _A eu on u 7 _ _ B Pse dom as_o doal 9 ocin AB AB 060 a_ 17 04 1 c udo ona s_a leovo alige D8 69 69 37 mo 4 5 9 Pse nas_ lcali rans nes fus phi _D _Z 0164 9 udo mon cov la_ Q8 76 99 agin AB 42 66 Pseu Pse 1 a s _ 0 u 6 dom onas domon asplenae_FJ3058018 Pseu as_ ii _vra 3 4 domo _ 4 AB0 835 38 a 4 n g n o 3 a v a 8 s e 1 0 Pseud rici_ 213 1996 09 omona _rhizosph nsis_AY Z76 97 66 a s_abie 2 tanip erae_A 9170951652 54 Pseudom 5267 onas_jap hila_AJ01Y 3 15 47 onica_A B12 04 Pseu

se

a s_ ctae_A P ud 3 B0213 P seud omon trema 78 40 Ps seud omon as_sa e_AJ492 82 3 6528 PPseeud omo as_c vastan oi_A 6 3759 se ud omo nas arica B _ o p 31 n 0 u c P a m 2 an a pay s d 2 ae_ 1402 21587 P eu om ona s_lin nab D84 0 P Ps se domona s_m i_AY ina_ 0

60

eu

ynxa

38

eu

eu ud do o na as_ fluo _AJ alis_ A F4 do om mo nas s_lu sal resc 936 053 m on na _c rid omo ens 933 28 on a s_ os a_ _ n as s_ pa tan AJ ii_A D840 _a tol lle tin 581 Y09 13 zo aa ron ii_A 99 1 52 to sii_ ian F3 9 8 fo A a rm F2 _A 744 an 55 Y0 72 s_ 33 91 52 D8 6 7 40 09

602

do

AF 3 98 a_ 667 98 ne Z7 Y50 2655 8 rra is_ _A 9 ite ph is Q6 53 ed ora raph s_D 168 _m lor ro phi _FJ 4 as ch hlo ra his 665 27 7 on s_ _c oro ap 40 395 m na as chl ror li_Z D8 5 1 do o on s_ hlo gda ns_ B02 188 eu om om ona s_c my trole is_A AB04 Pseudeud om ona as_a _tae dens hila_ 3 p Ps Pseuddommon onas _lun chro 09473 2 4577 Psseuudo dom onas s_psy gi_AF Y97095 P se seu dom ona _fra 70 50 _A P P eu om nas 7 nsis 4684 Ps seud domo moravie ensis_AF u P _ 7 g 29 Psemonas_umson _AM29356 4370 o s ii 55P8s 9eudomona s_mohn 468452 ud omona koreensis_AF 8 P se 3565 4 6 Pseudomonas_ kei_AM29 Pseudom onas_reine sis_AJ011507 Pseuddo s_vancouveren Pseu mona 259 Pseudomonas_jessenii_AF068 Pseudomonas_moorei_AM293566

eu

as as on n s m mo ona o o 7 d d m s 45 eu eu do ona s 64 Ps Ps eu m na s 0 F0 1 6_ Ps udo mo ona lis_A 9283 N _ 7_ e do m ta J4 61 BT TNN_ Ps seu udo rien a_A 0644 B T N8 P se _o drin AF 3 8 B T 9 P n a s ce n a _ 3 B TN 10 o as_ dri AF074 68 B T N d o m o n _ ce e _ 6 689 s 2 la B m F a u u A ig e o n 84 93 Psseuddomo as_m enneri_ AF0743 03 P seu mon s_br ardii_ 6 J5376 P u d o o n a g e ss A _ a c ti 7 1 7397Pseeudom onas_ proteoly 0 4 8 P s dom s_ lens_D 70 Pseuudomona s_mucido 025 ntha_D84 Pse mona 9 Pseuudomonas_s 166 57645 Pse anensis_AF0 4084 Pseudomonas_lib 87208 AY7 acis_ _pan onas dom 60 Pseu 4388 Pseudomonas_cichorii_Z76658

54 34 34 40 63

Ps

a P e m 0 4 PPs se udo on s_e ialis_ 92829 588 P seueud udo mo as_s xtrem AJ49 PsPseseu do omo mon nas_ imiae orien 2831 m t

1

Ps

0.01

5

P eu seu do do Ps Ps mon mo eu eu as na do m P do _p s_ Ps ona seu mo oha ma eu s_ do na n rin c d m s Ps om uat on _ta gen col eu on roc as ea si a_ do as ie _ n s_ A Pse _ m n b e D B udo P on ang eg orb ns Q 3 Pse mona seud as_g uilli ase ori_ is_ 33 010 om ui se ns A FJ 91 7 s_ ud Pse udo omona delhie onasneae pticais_EM1 42444 1 mon s_c nsis _p _A _X U7 14 8 Pseu a i t s _D el i M 4 9 9 52 13 r _ o dom 28 onas knackm nellol Q33_AM1 91895401287 20 Pseu _pan 1 ussii is_Z 915 14 10 domo ip 9 aten _AF0 7665 3 99534 n a s_ 23 s 9 6 1 Pseud nitroreduc is_EF4239489 67 omona e 4 84 55 19 Pseudom s_jinju ns_AM08401 onas_pa 58 8 e n 4 sis 73 chastrella 35 e_A B _A F 468448 Pseudomon as_duriflava 125366 2 _EU046271 96 Pseudomonas_luteola_ D84002 2 27 Pseudomonas_psychrotolerans_AJ575816 1 44 018 _D84 orans oleov 99 Pseudomonas_ 06 genes_D840 ali alc s_ 9 na 52 4 mo Pseudo 1 U1433 1 ulinigri_E 63 onas_sab gia_EU88898105 Pseudom 6 46 la e 8 p 2 _ U s a 7 E 7 64 2 is_ mon 3 0 9 Pseudo xinjiangens Q0886640 1 1988 16 onas_ nensis_D B02138AY69116 m o d 9 u e 3 _A nsis_ U01 0236996 m P se a ia n x _ e 4 04 4 8 2 520 e s _ g a 8 s in o n t n o in D dom rtuc _argen vesce nea_ _AB8400268 29 2289 e p _ 6 i 4 a 8 Pseu s a as fla m lv D 7 3 5 mon domon nas_ _stra s_fu ans_ AF0 10 44 457 udo o a bit lii_ EU 06 9 20 u s e m n a s Pse o P on mo iha se is_ F 00 0 ud Pse udom eudo oryz mos ens ilii_AAB 84 e Ps s_ s_ n te a_ _D Ps na na iwa on id da mo mo ta m sic ti do udo as_ as_ los _pu u e n n g Ps Pse mo mo co nas o o le ud ud _p mo e o P s P se n a s u d o se m P do eu Ps

6454

7 314 35 8 3 Y 95 is_A 684 68 25 807 _otitid _X06 Z766945295 8 4896 96 7 1 li_X _ s a _co mona ginos rans B18302264 9471 ella do aeru ovo ns_A_AF _U F0 69 91 Shig A seu in P hia/ as_ res ifige dica rica ri_ 770 06 n heric mo nas_ azot s_in alea utze AY Y77 Esc o b st _ udo a s_ m Pse eudo ona mon as_ as_ etis is_A n Ps udom udo mo on seg get e se do om s_ se P Ps e u u d n a s_ Ps Pse mo na do mo eu do P s eu Ps

Ps

63 Pseudomonas_marginalis_Z76663 50 Pseudom 27 _rhodesiae_AF064459 Pseudomoonas nas_ 16 Pse omonas_vgrimontii_AF268029 Pseuud eronii_AF0 1236 P d omo 6446 882 Psseudomo nas_extrem eudo australis 0 n a s_ a 2 7 P se m n _ A J5 8 u ta o P 3501 46 P seuddomon nas_me rctica_AJ 78 53760 Psseeudoomon as_po ridiana 1 238 6 _ a a A J5 3 7 Ps udo mon s_triv e_AJ4 7

S3 of S18

Mar. Drugs 2016, 14, 83

1 Psy BTN1

95

23

ct e

te

r_o

hro

r

k

bac

b chro

sy 85 P

Psych

ts ho

ro

te

a

ke

ns

is

ti_A r_lu

9 B0 _A

J4 3

47

082

8

us_ ritim _ma cter A ticus_

r_arc bacte

94

AJ6

092

Y 4448

BTN24_

19

19

0

42259 r_AY8

us_

r_cele robacte Psych

sals ter_ bac chro Psy

Psychrobacter_

04 391 A J5

M1 65 e_F

oca tiph r_lu cte rob a

ych Ps

namhaensis_AY7 228

05

94 09 J3 s_ A

nu ma ri ub r_s

cte ba

59

29

35

41 99 J3 0

_A

co la

r_ m ar in

ac te

hro

BTN22_Psychrobacter

yc

Ps

yc

hro

Ps

ba

ct e

yc

yc

hr ob

82

c

94

J _A

3

88 16 28

21

chr oba

hr ob

25 34 31 AJ 5 68 is_ 60 ar Y6 im _A niv tis r_ en te l ol ac ha yo ob r hr _c r yc cte Ps ba ter bac hro yc hr o c Ps Psy N3 BT cter 52 r oba h c y 5 Ps B TN r robacte _Psych BTN12

2 31

27

c la _g er

45

hrob

Psy

16

Ps

36

a ol

95 Psyc

59

16 52

56

51

BTN20B_Psychrobacter

94

ac

te

r_ p

ac

te

ro t

eo

Psychro

acte

cte

r_p

r_ p

r_a q

isc

bacter

r_va ll

is_A

uat icu

at o

rii_

s_A

AB

ac

J5 8 4

45

37

ific

en

lyt ic

us

_A

J2

72

30

sis _A

B0 1

832

J58

483

3

00

60

72

22

chrobacter

580

95

s riu

Ps yc hr ob

o hr

yc Ps

21

44

36

0 J6 _A

Ps

66

y

Ps

ac

99

b ro ch

41

12

t er

in

t

yc

73

a r_

92

ob hr

te ac

96

yc

a

yc

yc

ba hro

30

48

71

Y _A

t en lim

8 N1

Ps

99

bac

la

60

76

ac

ico

58

46

72

cter roba

igid

28

b hro

_fr t er

15

1 B TN

s 7_P

BT

s _P

c ba

BTN20A_Psy

sy BTN14 P

ro ych

AJ

89

0.02

1

er chrobact

21 B TN

_ sus

3 J4 43

t er

3

BTN19_Psychrobacter

ch _Psy

42

AB

ac

Y6

AJ

_AF44120

no

ro b

s_

A is_

ne

725

are s_ al i

tivo

X 80

er_

s_A

on

ter_jeotgali

Ps

ec

sy

ch

riu

_ coli

a ct

m ul

_P

vig e

39 87 ran 1 s_A J 6 095 rii_EU 5 9397 5 18

Psychrobac

Ps

a r_f

r_ fu l

BTN15_P

lla_

rob

p r_

te

N2 3

cte r_c iba

ura

ct e

estua

ba

ter_a

bac ter_

te ac

robac

ba

hro

Psych

BT

ac

BTN13 Psychrobacter

e Shig ych yc

Psy chro

hr ob

sychroba cter

hia/ heric Esc Ps Ps

Ps ych ro

yc

ob hr yc Ps

Ps

chrob acter B TN 2P syc Ps hr o ych bac ter rob ac Ps t er yc _a hr qu ob im ac ari BT t er s_ N1 AY _fo 72 6_ zii 28 _A Ps 04 J4 yc 30 hr 82 ob 7 ac te r

S4 of S18

57

3

(C) Figure S1. Phylogenetic trees (using the complete deletion option) of the Arthrobacter (A); Pseudomonas (B) and Psychrobater (C) genera.

Mar. Drugs 2016, 14, 83

S5 of S18

MayV-00635.10.1.1r UserName Aberdeen

17000000

SampleName BTN1-C8

16000000 15000000 14000000 13000000 12000000 11000000 10000000 9000000 8000000 7000000 6000000 5000000 4000000 3000000 2000000 1000000

-1000000

6.00

42.47

7.15

5.38

2.54

2.69

1.38

1.22

1.36

0

6.0 5.8 5.6 5.4 5.2 5.0 4.8 4.6 4.4 4.2 4.0 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6 1.4 1.2 1.0 0.8 0.6 0.4 0.2 f1 (ppm)

Figure S2. 1H-NMR spectrum of compound 1 in CD3OD at 600 MHz.

10

MayV-00635.12.1.2rr UserName Aberdeen SampleName BTN1-C8 eHSQCedSolvent: MeOH

20

30

40

60

70

80

90

100

5.4

5.2

5.0

4.8

4.6

4.4

4.2

4.0

3.8

3.6

3.4

3.2 3.0 f2 (ppm)

2.8

2.6

2.4

2.2

2.0

1.8

1.6

1.4

1.2

Figure S3. Edited-HSQC spectrum of compound 1 in CD3OD at 600 MHz.

1.0

0.8

f1 (ppm)

50

Mar. Drugs 2016, 14, 83

S6 of S18

MayV-00635.11.1.2rr UserName Aberdeen

1.0

SampleName BTN1-C8 Experiment: eCOSY Solvent: MeOH 1.5

2.0

3.0

3.5

4.0

4.5

5.0

5.5

5.8 5.6 5.4 5.2 5.0 4.8 4.6 4.4 4.2 4.0 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6 1.4 1.2 1.0 0.8 f2 (ppm)

Figure S4. COSY NMR spectrum of compound 1 in CD3OD at 600 MHz.

0

MayV-00635.13.1.2rr UserName Aberdeen

10

SampleName BTN1-C8 Experiment: eHMBC_NUS.MeOH

20 30 40 50 60

80 90 100 110 120 130 140 150 160 170 5.6

5.4

5.2

5.0

4.8

4.6

4.4

4.2

4.0

3.8

3.6

3.4

3.2 3.0 f2 (ppm)

2.8

2.6

2.4

2.2

2.0

1.8

1.6

1.4

1.2

Figure S5. HMBC spectrum of compound 1 in CD3OD at 600 MHz.

1.0

0.8

0.6

f1 (ppm)

70

f1 (ppm)

2.5

Mar. Drugs 2016, 14, 83

S7 of S18

BTN1-C8_DMSO.37.fid 12000 11000 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000 0

6.2 6.0 5.8 5.6 5.4 5.2 5.0 4.8 4.6 4.4 4.2 4.0 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6 1.4 1.2 1.0 0.8 0.6 0.4 0.2 0.0 f1 (ppm)

Figure S6. 1H-NMR spectrum of compound 1 in DMSO-d6 at 400 MHz. MayV-00633.10.fid UserName Aberdeen BTN1-C6

19000 18000 17000 16000 15000 14000 13000 12000 11000 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000

8.0

7.5

7.0

6.5

6.0

5.5

5.0

4.5

4.0 f1 (ppm)

3.5

3.0

2.5

2.0

1.5

-1000

6.00

21.72

2.31

1.80 2.18 1.13 1.15 2.18

1.11

1.03 2.29

1.18

1.16 1.25 1.27

0

1.0

Figure S7. 1H-NMR spectrum of compound 2 in CD3OD at 600 MHz.

0.5

0.0

Mar. Drugs 2016, 14, 83

S8 of S18

MayV-00633.12.ser UserName Aberdeen

10

SampleName BTN1-C6 Experiment: eHSQCed_NUS.

20 30 40 50

70 80

f1 (ppm)

60

90 100 110 120 130 140 5.8 5.6 5.4 5.2 5.0 4.8 4.6 4.4 4.2 4.0 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6 1.4 1.2 1.0 0.8 0.6 0.4 0.2 f2 (ppm)

Figure S8. Edited-HSQC NMR spectrum of compound 2 in CD3OD.

MayV-00633.11.ser UserName Aberdeen SampleName BTN1-C6 COSY

1.0

1.5

2.0

2.5

3.5

4.0

4.5

5.0

5.5

6.0

6.0 5.8 5.6 5.4 5.2 5.0 4.8 4.6 4.4 4.2 4.0 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6 1.4 1.2 1.0 0.8 f2 (ppm)

Figure S9. COSY NMR spectrum of compound 2 in CD3OD.

f1 (ppm)

3.0

Mar. Drugs 2016, 14, 83

S9 of S18

0

MayV-00633.13.1.2rr _BTN1-C6_180515_1H-2DCOSY-2DHSQCed-2DHMBC_av600_MayV-00633

10 20 30 40 50 60 70

90 100 110 120 130 140 150 160 170 180 6.0

4.5

5.0

5.5

4.0

3.5 f2 (ppm)

2.0

2.5

3.0

1.0

1.5

0.5

Figure S10. HMBC NMR spectrum of compound 2 in CD3OD. BTN1-C6_DMSO.38.fid

12000 11000 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000 0

8.0

7.5

7.0

6.5

6.0

5.5

5.0

4.5

4.0 f1 (ppm)

3.5

3.0

2.5

2.0

1.5

1.0

Figure S11. 1H-NMR spectrum of compound 2 in DMSO-d6 at 400 MHz.

0.5

0.0

f1 (ppm)

80

Mar. Drugs 2016, 14, 83

S10 of S18

Figure S12. 2D ROESY spectrum of compound 2 in DMSO-d6 with Rhamnose protons labelled.

C5H12

C7H15

O O

H

O OH

O

H H

O HO

HO H

H OH

COSY ROE

Figure S13. Compound 2 showing selected ROE correlations of the rhamnose moiety and COSY correlations showing position of unsaturation of the lipid chain.

Mar. Drugs 2016, 14, 83

S11 of S18

MayV-00637.14.1.1r SampleName BTN1-C10 Experiment: eSpresat Solvent: MeOH

3000000 2800000 2600000 2400000 2200000 2000000 1800000 1600000 1400000 1200000 1000000 800000 600000 400000 200000 0 -200000

5.8

5.4

4.6

5.0

4.2

3.8

2.6 3.0 f1 (ppm)

3.4

1.8

2.2

1.4

1.0

0.6

0.2

-0.2

Figure S14. 1H-NMR spectrum of compound 3 in CD3OD at 600 MHz.

10

MayV-00637.12.1.2rr SampleName BTN1-C10 Experiment: eHSQCed_NUS.highres Solvent: MeOH Holder: 11 Group

20 30 40 50

70 80 90 100 110 120 130 5.5

5.0

4.5

4.0

3.5

3.0

2.5

2.0

1.5

f2 (ppm)

Figure S15. Edited-HSQC NMR spectrum of compound 3 in CD3OD.

1.0

f1 (ppm)

60

Mar. Drugs 2016, 14, 83

S12 of S18

MayV-00637.11.1.2rr SampleName BTN1-C10 Experiment: eCOSY Solvent: MeOH Holder:

1.0

1.5

2.0

3.0

3.5

f1 (ppm)

2.5

4.0

4.5

5.0

5.5

5.8 5.6 5.4 5.2 5.0 4.8 4.6 4.4 4.2 4.0 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6 1.4 1.2 1.0 0.8 f2 (ppm)

Figure S16. COSY NMR spectrum of compound 3 in CD3OD.

MayV-00637.13.1.2rr SampleName BTN1-C10 Experiment: eHMBC_NUS.highres Solvent: MeOH

20 30 40 50 60 70

90 100 110 120 130 140 150 160 170 180 5.8 5.6 5.4 5.2 5.0 4.8 4.6 4.4 4.2 4.0 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6 1.4 1.2 1.0 0.8 0.6 0.4 f2 (ppm)

Figure S17. HMBC NMR spectrum of compound 3 in CD3OD.

f1 (ppm)

80

Mar. Drugs 2016, 14, 83

S13 of S18

Retention Time: 21.149

Ion Mode: ESI+ 555.351 555.351

22 20 18 16 14 12 10 556.356 556.356

8 6 4 2

554.783 554.783

ESI+

554.406 554.406 554.083 554.083

557.359 556.624 556.624

555.920 555.920

557.632 557.632 558.359 558.359

0 554.0

554.5

555.0

555.5

556.0

556.5

557.0

557.5

558.0

Figure S18. HRESIMS spectrum (M + Na)+ of compound 1.

Figure S19. ESI MS/MS data of compound 1.

558.5

m/z

Mar. Drugs 2016, 14, 83

S14 of S18

Retention Time: 20.240

Ion Mode: ESI+

70 65 60 55 553.337 553.337

50 45 40 35 30 25 20

554.339 554.339 15 10 5

ESI+

555.341 555.341 554.603 554.603

553.148 553.148

552.073 552.073 551.329 551.329

552.770 552.770

551.5

552.5

555.608 555.608 556.346 556.346

0 552.0

553.0

553.5

554.0

554.5

555.0

555.5

Figure S20. HRESIMS spectrum (M + Na)+ of compound 2.

Figure S21. ESI MS/MS data of compound 2.

556.0

556.5

m/z

Mar. Drugs 2016, 14, 83

S15 of S18 581.368 581.368

Retention Time: 21.698

Ion Mode: ESI+

96 88 80 72 64 56 48 40 582.370 582.370 32 24 16 8

ESI+ 580.352 580.352

583.374 583.374 581.977 581.977 581.571 581.571

580.757 580.757

582.983 582.983

583.742 583.742

584.376 584.376

0 580.5

581.0

581.5

582.0

582.5

583.0

583.5

Figure S22. HRESIMS spectrum (M + Na)+ of compound 3.

Figure S23. ESI MS/MS data for compound 3.

584.0

m/z

Mar. Drugs 2016, 14, 83

S16 of S18

Table S1. Cross- streaking experiments of BTN isolates against a wide panel of Bcc strains. The Bcc strains highlighted in red correspond to type strains.Symbols: +, growth; ±, reduced growth; -, no growth; C+, positive controls, i.e., Bcc strains grown in the absence of the tester strain(s). Abbreviations: Ps, Pseudomonas; Ar, Arthrobacter; CF, Cystic Fibrosis; Env; environmental. Bcc Species

B. cepacia

B. multivorans

B. cenocepacia

Source

Strain

C+

CF

FCF 1

CF

FCF 2

CF

Ps

Psychrobacter BTN21 BTN24 BTN23

BTN5

Ar

BTN1

BTN2

BTN15

BTN3

BTN19

BTN20B

BTN12 BTN14 BTN17 BTN16 BTN18 BTN11 BTN13

BTN22

BTN4

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

FCF3

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Env

LMG 2161

+

-

-

-

-

±

-

-

-

-

-

-

-

-

-

CF

FCF 5

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 6

+

-

-

-

-

±

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 7

+

-

-

-

-

±

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 8

+

-

-

-

-

±

-

-

-

-

-

-

CF

FCF 9

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 10

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 11

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

LMG 18822

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Env

LMG 17588

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 12

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 13

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 14

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 15

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 16

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 17

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 18

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 19

+

±

-

-

±

-

-

-

-

±

-

±

±

-

±

-

±

-

-

±

CF

FCF 20

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 21

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 22

+

-

-

±

-

-

±

±

-

-

±

-

-

-

-

-

-

-

-

-

CF

FCF 23

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 24

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 25

+

±

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

±

-

CF

FCF 26

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Mar. Drugs 2016, 14, 83

B. stabilis B. vietnamensis B. dolosa B. ambifaria B. anthina

B. pyrrocinia

S17 of S18

CF

FCF 27

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 28

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 29

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 30

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 31

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

LMG 16654

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

C 5424

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

CEP 511

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Env

MVPC 1/16

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Env

MVPC 1/73

+

-

-

-

-

-

-

-

-

±

-

-

-

-

-

-

-

-

-

-

CF

LMG 24506

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Env

LMG 19230

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Env

LMG 19240

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 32

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 33

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 34

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 36

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 37

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 38

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 39

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

LMG 21462

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 40

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 41

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 42

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

LMG 18941

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

LMG 18942

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Env

MCI 7

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

LMG 19467

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Env

LMG 16670

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

LMG 20983

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 43

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 44

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

CF

FCF 45

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Mar. Drugs 2016, 14, 83

B. lata

S18 of S18

CF

FCF 46

+

-

-

-

-

-

-

-

-

Env

MVPC 1/26

+

-

-

-

-

-

-

-

Env

MVPC 2/77

+

-

-

-

-

-

-

-

CF

LMG 21824

+

-

-

-

-

-

±

CF

LSED 4

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

±

-

±

-

-

-

-

-

-

-

-

-

-

±

-

±

-

-

-

-

-

-

±

-

-

±

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Env

LMG 6991

+

-

-

-

-

±

±

±

-

±

-

-

-

-

±

-

-

-

-

±

B. ambifaria

Env

LMG 19182

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

±

-

-

-

B. anthina

Env

LMG 20980

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

B. cenocepacia

CF

LMG 16656

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

B. cepacia

Env

LMG 1222

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

B. contaminas

AI

LMG 23361

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

B. diffusa

CF

LMG 24065

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

B. dolosa

CF

LMG 18943

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

B. lata

Env

LMG 22485

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

B. latens

CF

LMG 24064

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

B. metallica

CF

LMG 24068

+

-

-

+

-

-

±

±

±

±

±

+

-

-

-

-

-

-

-

-

B. multivorans

CF

LMG 13010

+

-

-

±

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

B. pseudomultivorans

CF

LMG 26883

+

-

-

±

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

B. pyrrocinia

Env

LMG 14191

+

-

-

+

-

-

-

-

±

±

-

+

-

-

-

-

-

-

-

-

B. seminalis

CF

LMG 24067

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

B. stabilis

CF

LMG 14294

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

B. uborrensis

Env

LMG 20358

+

-

-

±

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

B. vietnamensis

Env

LMG10929

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-