SUPPORTING INFORMATION
(a)
9.0E+07
1-K
6.0E+07
6G-K
Ion intensity
1.2E+08
3.0E+07
0.0E+00
0 1 2 3 4 5 6 7 8 9 10
d
4.5E+05
6-K
Ion intensity
6.0E+05
(b)
3.0E+05
1.5E+05
0.0E+00
3.9E+06
(c)
6-K
2.6E+06
1.3E+06
1-K
Ion intensity
0 1 2 3 4 5 6 7 8 9 10
d
5.2E+06
0.0E+00
0 1 2 3 4 5 6 7 8 9 10
Time (min)
Figure S1. (a) Extracted ion profile (XIC) of the 6-SFT reaction products incubated with 1-kestotriose and [UL-13C6
fru] sucrose at m/z 503, corresponding to native oligosaccharides with a degree of polymerization (DP) of 3. (b) XIC
of the 6-SFT reaction products at m/z 509, corresponding to DP 3 oligosaccharides with one UL-13C6 fructose unit.
(c) XIC of the 6-SFT reaction products at m/z 515, corresponding to DP 3 oligosaccharides with two UL-13C6
fructose units.
1&6-KT
0.0E+00
1,1-KT
6G,6-KT
2.0E+07
(a)
1,6-KT
4.0E+07
6,6-KT
Ion intensity
6.0E+07
0 1 2 3 4 5 6 7 8 9 10
(b)
6G,6-KT
1.0E+07
1&6-KT
Ion intensity
2.0E+07
1,6-KT
Time (min)
3.0E+07
0.0E+00
0 1 2 3 4 5 6 7 8 9 10
4.0E+05
2.0E+05
0.0E+00
Time (min)
(c)
6,6-KT
1&6-KT
1,6-KT
Ion intensity
6.0E+05
0 1 2 3 4 5 6 7 8 9 10
Time (min)
Figure S2. (a) XIC of the 6-SFT reaction products at m/z 665, corresponding to native DP 4 oligosaccharides, in the
6-SFT reaction mixture with 1-K and unlabeled sucrose. (b) XIC of the 6-SFT reaction products at m/z 671,
corresponding to DP 4 oligosaccharides with one UL-13C6 fructose unit, in the 6-SFT reaction mixture with 1-K and
UL-labeled sucrose. (c) XIC of the 6-SFT reaction products at m/z 683, corresponding to DP 4 oligosaccharides with
three UL-13C6 fructose units, in the 6-SFT reaction mixture with 1-K and UL-labeled sucrose.
40,000
30,000
20,000
221
X1
10,000
341
*
(a)
40,000
Intensity
50,000
Intensity
389
383
X2
30,000
X2
20,000
227
10,000
0
(b)
*
347
341
250
350
m/z
X1
0
150
250
350
m/z
450
550
150
450
550
Figure S3. (a) Negative electrospray MS2 spectrum (m/z 665) of 6G,6-kestotetraose. (b) MS2 spectrum (m/z 671) of
6G,6-kestotetraose with one UL-13C6 fructose unit.
Ion intensity
1.2E+07
A
(a)
9.0E+06
6.0E+06
3.0E+06
B
0.0E+00
0 1 2 3 4 5 6 7 8 9 10
Ion intensity
8.0E+06
ATime (min)
(b)
6.0E+06
4.0E+06
2.0E+06
B
0.0E+00
0 1 2 3 4 5 6 7 8 9 10
Time (min)
Figure S4. (a) XIC of the 6-SFT reaction products at m/z 827, corresponding to native DP 5 oligosaccharides. (b)
XIC of the 6-SFT reaction products at m/z 839, corresponding to DP 5 oligosaccharides with two UL-13C6 fructose
units.
Ion intensity
6.0E+05
(a)
C
4.0E+05
2.0E+05
B
A
D
0.0E+00
0 1 2 3 4 5 6 7 8 9 10
Time (min)
Ion intensity
1.2E+06
B
8.0E+05
4.0E+05
A
(b)
C
D
0.0E+00
0 1 2 3 4 5 6 7 8 9 10
Time (min)
Figure S5. (a) XIC of the 6-SFT reaction products at m/z 989, corresponding to native DP 6
oligosaccharides. (b) XIC of the 6-SFT reaction products at m/z 992, corresponding to DP 6
oligosaccharides with three 13C1 fructose units.
Figure S6. Graminan synthesis by 6-SFT incubated with 1-kestotriose and unlabeled sucrose after 0 h, 72 h, 120 h
and 175 h incubation, respectively.
0.032353446
0.553436451
1.007632375
1.456338059
1.901579969
2.355966299
2.811565538
3.270488674
3.726986366
4.183882054
4.640823439
5.098392682
5.55796997
6.015798156
6.473252087
6.931036259
7.388825872
7.846689991
8.306316895
8.764098415
9.221593176
9.679723627
70000000
(a)
(1)
0.032085943
0.546918418
1.002970304
1.454818208
6-K
1.91329286
1&6-KT2.371468994
2.828473744
3.284663457
3.733894725
4.191767838
4.650879935 6G-K
5.109061266
5.567206057
6.024814745
6.482148446
6.940223344
7.400068262
7.858050599
8.315321668
1,1-KT
8.739458242
9.195228334
9.656687882
(2)
(3)
0
2
3
4
5
6
7
8
9
Time (min)
(b)
6,6-KT
1
2
10000000
0
60000000
50000000
40000000
30000000
1,1-KT
1,6-KT
(5)
0.0E+00
0
20000000
10
(4)
6,6-KT
Ion intensity (au)
6.0E+07
1
40000000
30000000
6.0E+07
0.0E+00
60000000
50000000
6G-K
1.2E+08
1-K
6-K
Ion intensity (au)
1.8E+08
3
4
5
6
7
8
9
20000000
10000000
(6)
0
10
Time (min)
Figure S7. (a) Extracted ion profile (XIC) at m/z 503, corresponding to DP 3 fructans, of a 6-kestotriose (6-K)
standard (1), 6G-kestotriose (6G-K) standard (2) and the 6-SFT reaction mixture (3) containing mainly 1-kestotriose
(1-K). (b) XIC at m/z 665, corresponding to DP 4 fructans, of a 1,1-kestotetraose (1,1-KT) standard (4), 6,6kestotetraose (6,6-KT) standard (5) and the 6-SFT reaction mixture (6) containing mainly 1&6-kestotetraose (1&6KT) and 1,6-kestotetraose (1,6-KT) as DP 4 fructans.
Table S1. Overview of the observed 6-SFT products, their retention times (tr), ion intensities and
the ion intensities of the corresponding formate adduct. Values are listed for the reaction products
after incubation with 1-kestotriose and unlabeled sucrose (A), [1-13C1 fru] sucrose (B) or [UL13
C6 fru] sucrose (C)
Table S1.A
Fructan
1&6-KT
1,6-KT
1&6,6-KP/1,6&6-KP
1,6,6-KP
6&1,6,6-KH
1,6&6,6-KH
1&6,6,6-KH
1,6,6,6-KH
tr (min)
2.4
3.7
2.7
3.5
2.9
3.1
3.4
4.1
m/z
[M-H]665.21
665.21
827.27
827.27
989.32
989.32
989.32
989.32
ion intensity
[M-H]1.96E+07
5.95E+06
5.32E+06
3.96E+05
4.41E+04
6.25E+04
2.33E+05
4.89E+04
ion intensity
[M+CHO2]5.12E+07
2.18E+06
1.94E+06
1.39E+05
3.91E+04
0.00E+00
9.35E+04
0.00E+00
ion intensity
[M-H]–
3.08E+07
6.59E+06
8.11E+06
6.55E+05
1.20E+05
1.09E+05
5.36E+05
9.90E+04
ion intensity
[M+CHO2]–
8.36E+07
2.35E+06
3.08E+06
2.33E+05
1.12E+05
3.40E+02
2.36E+05
2.78E+02
Table S1.B
Fructan
13
[1- C1 fru] 1&6-KT
[1-13C1 fru] 1,6-KT
[1-13C1 fru]2 1&6,6-KP/1,6&6-KP
[1-13C1 fru]2 1,6,6-KP
[1-13C1 fru]3 6&1,6,6-KH
[1-13C1 fru]3 1,6&6,6-KH
[1-13C1 fru]3 1&6,6,6-KH
[1-13C1 fru]3 1,6,6,6-KH
tr (min)
2.4
3.7
2.7
3.5
2.9
3.1
3.4
4.1
m/z
[M-H]–
666.21
666.21
829.27
829.27
992.32
992.32
992.32
992.32
Table S1.C
m/z
Fructan
tr (min)
[M–H]–
[UL-13C6 fru] 1&6-KT
2.4
671.21
13
[UL- C6 fru] 1,6-KT
3.7
671.21
[UL-13C6 fru]2 1&6,6-KP/1,6&6-KP
2.7
839.27
13
[UL- C6 fru]2 1,6,6-KP
3.5
839.27
13
[UL- C6 fru]3 6&1,6,6-KH
2.9
1007.32
13
[UL- C6 fru]3 1,6&6,6-KH
3.1
1007.32
13
[UL- C6 fru]3 1&6,6,6-KH
3.4
1007.32
13
[UL- C6 fru]3 1,6,6,6-KH
4.1
1007.32
*Not informative because of coelution with 1-kestotriose adducts
ion intensity
[M–H]–
1.30E+07
4.29E+06
4.33E+06
4.41E+05
8.19E+04
*
*
7.84E+05
ion intensity
[M+CHO2]–
3.71E+07
1.70E+06
1.72E+06
1.52E+05
7.98E+04
*
*
1.72E+04