Journal of American Society of Mass Spectrometry

ELECTRONIC SUPPLEMENTARY MATERIAL

Lipopeptides from the Banyan Endophyte, Bacillus Subtilis K1: Mass Spectrometric Characterization of a Library of Fengycins

Journal of American Society of Mass spectrometry

Khyati V. Pathaka,b, Haresh Kehariab, Kallol Guptaa, Suman S. Thakura, Padmanabhan Balarama

a Molecular Biophysics Unit, Indian Institute of Science, Bangalore 560012, India

b BRDSchool of Biosciences, SardarPatelUniversity,Vallabh Vidyanagar, Anand 388120, India

Address reprint requests to

Prof. Padmanabhan Balaram

Molecular Biophysics Unit, Indian Institute of Science,

Bangalore 560012, India. Fax: 91-80-236060683/91-23600535.

Phone: 91-80-22932741. E-mail:

Table of content

S.1Figure S1. ESI-MS/MS of protonated precursor ions at a) m/z 1447.8 (cyclic) b) m/z 1465.8 (linearized) from HPLC fraction P19.

S.2Figure S2. ESI MS spectrum of benzyl esters of fengycins in HPLC peaks a)P14 and b) P19. (Insets) Observed and calculated isotope distributions of representative ion clusters at a) m/z 1630.1; m/z 1721.1 and b) m/z 1669.9; 1762.9.

S.3 Figure S3. ESI-MS spectra of control and esterification reaction of iturin As in HPLC peak P4 using benzyl alcohol/acetyl chloride (50:1 v/v) over a time period of 1 to 4 h.

S.4 Figure S4. MALDI-MS spectra of control and esterification reaction of fengycin in HPLC peak P17 using benzyl alcohol/acetyl chloride (50:1 v/v) over a time period of 1 to 10 h.

S.5Figure S5. MALDI-MS spectra of HPLC peak P17 control and P17 at pH2 after incubation for 15h.

S.6Figure S6. MALDI-MS of a cell suspension of B. subtilis K1. Fengycins ion clusterare expanded.

S.7Figure S7. MALDI-MS/MS of the protonated precursor ion at m/z 1467.5 from HPLC fraction P6.

S.8Table S1. Fengycins in crude extract of B. subtilis K1

S.1 Figure S1. ESI-MS/MS of protonated precursor ions at a) m/z 1447.8 (cyclic) b) m/z 1465.8 (linearized) from HPLC fraction P19.

S.3Figure S3. ESI-MS spectra of control and esterification reaction of iturin As in HPLC peak P4 using benzyl alcohol/acetyl chloride (50:1 v/v) over a time period of 1 to 4 h.

S.4 Figure S4. MALDI-MS spectra of control and esterification reaction of fengycin in HPLC peak P17 using benzyl alcohol/acetyl chloride (50:1 v/v) over a time period of 1 to 10 h.

S.5 Figure S5. MALDI-MS spectra of HPLC peak P17 control and P17 at pH2 after incubation for 15h.

S.6Figure S6. MALDI-MS of a cell suspension of B. subtilis K1. Fengycins ion clusterare expanded.

S.7 Figure S7. MALDI-MS/MS of the protonated precursor ion at m/z 1467.5 from HPLC fraction P6.

S.8Table S1. Fengycins in crude extract of B. subtilis K1

Peak
No. / No. / Mass
(m/z) / Typical product Ions
(MS/MS
of Intact Fengycins) / Sequence
(obtained through saponification) / Identification
Of
Fengycin / Reference
6 / 1
2 / 1453.5
1454.5 / - / R’(C15)EOrnYTEAPQYV
R’(C15)EOrnYTEAPEYV / Linear (C15) Fengycin A2
Linear (C15)Fengycin A’2 / This work(New)
This work(New)
6 / 3
4 / 1453.5
1454.5 / - / R’(C14)EOrnYTEAPQYI
R’(C14)EOrnYTEAPEYI / Linear (C14) Fengycin A1
Linear (C14) Fengycin A’1 / This work
This work (New)
6 / 5
6 / 1467.5
1468.5 / - / R’(C14)EOrnYTEVPQYV
R’(C14)EOrnYTEVPEYV / Linear (C14) Fengycin B2
Linear (C14) Fengycin B’2 / This work(New)
This work(New)
6 / 7
8 / 1481.5
1482.5 / - / R’(C14)EOrnYTEVPQYI
R’(C14)EOrnYTEVPEYI / Linear (C14) Fengycin B1
Linear (C14) Fengycin B’1 / This work
This work(New)
8 / 9 / 1421.8 / 952,1066 / R’(C14)EOrnYTEAPQYV / (C14) Fengycin A2 / This work(New)
19 / 10 / 1432.8 / 952,1066 / R’(C14)EOrnYTEAPQYV / (C15) Fengycin A2a / This work(New)
8 / 11
12 / 1435.8
1436.8 / 952,1066
953,1067 / R’(C15)EOrnYTEAPQYV
R’(C15)EOrnYTEAPEYV / (C15) Fengycin A2
(C15) Fengycin A’2 / This work(New)
This work(New)
9 / 13 / 1435.8 / 966,1080 / R’(C14)EOrnYTEAPQYI / (C14) Fengycin A1 / This work, [23]
19 / 14
15 / 1447.8
1448.8 / 966,1080
967,1081 / R’(C-15)EOrnYTEAPQYI
R’(C-15)EOrnYTEAPEYI / (C15) Fengycin A1 a
(C15) Fengycin A’1 a / This work, [11]
This work(New)
14 / 16
17 / 1449.8
1450.8 / 952,1066
953,1067 / R’(C16)EOrnYTEAPQYV
R’(C16)EOrnYTEAPEYV / (C16) Fengycin A2
(C16) Fengycin A’2 / This work(New)
This work(New)
8 / 18
19 / 1449.8
1450.8 / 966,1080
967,1081 / R’(C15)EOrnYTEAPQYI
R’(C15)EOrnYTEAPEYI / (C15) Fengycin A1
(C15) Fengycin A’1 / This work, [23]
This work(New)
9 / 20
21 / 1449.8
1450.8 / 980,1094
981,1095 / R’(C14)EOrnYTEVPQYV
R’(C14)EOrnYTEVPEYV / (C14) Fengycin B2
(C14) Fengycin B’2 / This work(New)
This work(New)
19 / 22
23 / 1461.8
1462.8 / 980,1094
981,1095 / R’(C-15)EOrnYTEVPQYV
R’(C-15)EOrnYTEVPEYV / (C15) Fengycin B2a
(C15) Fengycin B’2a / This work (New)
This work (New)
14 / 24 / 1464.8 / 952,1066 / R’(C17)EOrnYTEAPEYV / (C17) Fengycin A’2 / This work (New)
14 / 25 / 1464.8 / 966,1080 / R’(C16)EOrnYTEAPEYI / (C16) Fengycin A’1 / This work (New)
15 / 26 / 1463.8 / 952,1066 / R’(C17)EOrnYTEAPQYV / (C17) Fengycin A2 / This work, [12] (proposed)
15 / 27 / 1463.8 / 966,1080 / R’(C16)EOrnYTEAPQYI / (C16) Fengycin A1 / This work,[23]
13 / 28
29 / 1463.8
1464.8 / 980,1094
981,1095 / R’(C15)EOrnYTEVPQYV
R’(C15)EOrnYTEVPEYV / (C15) Fengycin B2
(C15) Fengycin B’2 / This work, [12] (proposed)
This work (New)
19 / 30
31 / 1463.8
1464.8 / 994,1108
995,1109 / R’(C14)EOrnYTEVPQYI
R’(C14)EOrnYTEVPEYI / (C14) Fengycin B1
(C14) Fengycin B’1 / This work
This work (New)
19 / 32 / 1475.8 / 994,1108 / R’(C15)EOrnYTEVPQYI / (C15) FengycinB1a / This work
17 / 33 / 1477.8 / 966,1080 / R’(C17)EOrnYTEAPQYI / (C17) Fengycin A1 / This work, [23]
16 / 34
35 / 1477.8
1478.8 / 980,1094
981,1095 / R’(C16)EOrnYTEVPQYV
R’(C16)EOrnYTEVPEYV / (C16) Fengycin B2
(C16) Fengycin B’2 / This work(New)
15 / 36
37 / 1477.8
1478.8 / 994,1108
995,1109 / R’(C15)EOrnYTEVPQYI
R’(C15)EOrnYTEVPEYI / (C15) FengycinB1
(C15) FengycinB’1 / This work, [23]
This work(New)
19 / 38
39 / 1481.8
1482.8 / - / R’(C17)EOrnYTEAPQYV
R’(C17)EOrnYTEAPEYV / Linear (C16) Fengycin A2
Linear (C16) Fengycin A’2 / This work (New)
This work (New)
19 / 40
41 / 1481.8
1482.8 / - / R’(C16)EOrnYTEAPQYI
R’(C16)EOrnYTEAPEYI / Linear (C15) Fengycin A1
Linear (C15) Fengycin A’1 / This work
This work (New)
19 / 42 / 1489.8 / 1008,1122 / R’(C15)EOrnYTEIPQYI / (C15) Fengycin C1a / This work (New)
18 / 43
44 / 1491.8
1492.8 / 966,1080
967,1081 / R’(C-18)EOrnYTEVPQYV
R’(C-18)EOrnYTEVPEYV / (C18) Fengycin A1
(C18) Fengycin A’1 / This work, [11]
This work (New)
19 / 45
46 / 1491.8
1492.8 / 980,1094
981,1095 / R’(C-17)EOrnYTEVPQYV
R’(C-17)EOrnYTEVPEYV / (C17) Fengycin B2
(C17) Fengycin B’2 / This work(New)
17 / 47 / 1491.8 / 994,1108 / R’(C16) EOrnYTEVPQYI / (C16)Fengycin B1 / This work,[23]
19 / 48
49 / 1494.8
1495.8 / - / R’(C17) EOrnYTEAPQYI
R’(C17) EOrnYTEAPEYI / Linear (C17) Fengycin A1
Linear (C17) Fengycin A’1 / This work
This work(New)
19 / 50
51 / 1494.8
1495.8 / - / R’(C16)EOrnYTEVPQYV
R’(C16)EOrnYTEVPEYV / Linear (C16) Fengycin B2
Linear (C16) Fengycin B’2 / This work (New)
This work (New)
19 / 52
53 / 1505.8
1506.8 / 966,1080
967,1081 / R’(C19)EOrnYTEAPQYI
R’(C19)EOrnYTEAPEYI / (C19)Fengycin A1
(C19)Fengycin A’1 / This work
This work (New)
18 / 54 / 1505.8 / 980,1094 / R’(C18)EOrnYTEVPQYV / (C18)Fengycin B2 / This work(New)
19 / 55
56 / 1505.8
1506.8 / 994,1108
995,1109 / R’(C17)EOrnYTEVPQYI
R’(C17)EOrnYTEVPEYI / (C17)Fengycin B2
(C17)Fengycin B’2 / This work, [23]
This work(New)
19 / 57
58 / 1505.8
1506.8 / 1008,1122
1009,1123 / R’(C17)EOrnYTEIPQYI
R’(C17)EOrnYTEIPEYI / (C16)Fengycin C
(C16)Fengycin C’ / This work(New)
19 / 59
60 / 1509.8
1510.8 / - / R’(C17)EOrnYTEVPQYV
R’(C17)EOrnYTEVPEYV / Linear (C17)Fengycin B2
Linear (C17)Fengycin B’2 / This work (New)
19 / 61
62 / 1519.8
1520.8 / 980,1094
981,1095 / R’(C19)EOrnYTEVPQYV
R’(C19)EOrnYTEVPEYV / (C19)Fengycin B2
(C19)Fengycin B’2 / This work (New)
19 / 63
64 / 1519.8
1520.8 / 994,1108
995,1109 / R’(C18)EOrnYTEVPQYI
R’(C18)EOrnYTEVPEYI / (C18)Fengycin B1
(C18)Fengycin B’1 / This work, [23]
This work (New)
19 / 65
66 / 1519.8
1520.8 / 1008,1122
1009,1123 / R’(C17)EOrnYTEIPQYI
R’(C17)EOrnYTEIPEYI / (C17)Fengycin C
(C17)Fengycin C’ / This work (New)
19 / 67
68 / 1533.8
1534.8 / 980,1094
981,1095 / R’(C20)EOrnYTEVPQYV
R’(C20)EOrnYTEVPEYV / (C20)Fengycin B2
(C20)Fengycin B’2 / This work (New)
19 / 69
70 / 1533.8
1534.8 / 994,1108
995,1109 / R’(C19)EOrnYTEVPQYI
R’(C19)EOrnYTEVPEYI / (C19)Fengycin B1
(C19)Fengycin B’1 / This work
This work (New)
19 / 71
72 / 1533.8
1534.8 / 1008,1122
1009,1123 / R’(C18)EOrnYDEIPQYI
R’(C18)EOrnYDEIPEYI / (C18)Fengycin C
(C18)Fengycin C’ / This work(New)
19 / 73 / 1537.9 / - / R’(C18)EOrnYTEVPQYI / Linear (C18)Fengycin B1 / This work
19 / 74 / 1537.9 / - / R’(C17)EOrnYTEIPQYI / Linear (C17)Fengycin C / This work (New)
19 / 75
76 / 1547.8
1548.8 / 980,1094
981,1095 / R’(C21)EOrnYTEVPQYV
R’(C21)EOrnYTEVPEYV / (C21)Fengycin B2
(C21)Fengycin B’2 / This work (New)
19 / 78
79 / 1547.8
1548.8 / 994,1108
995,1095 / R’(C20)EOrnYTEVPQYI
R’(C20)EOrnYTEVPEYI / (C20)Fengycin B1
(C20)Fengycin B’1 / This work
This work (New)
19 / 81
81 / 1547.8
1548.8 / 1008,1122
1009,1123 / R’(C19)EOrnYTEIPQYI
R’(C19)EOrnYTEIPEYI / (C19)Fengycin C
(C19)Fengycin C’ / This work(New)

R’ =β-hydroxy fatty acid; a Mono unsaturated β-hydroxy fatty acid