SUPPLEMENTARY MATERIALS Catalytic activity of halohydrin ... · S6 3.2. Overlay of equatorial and...

S1

SUPPLEMENTARY MATERIALS

Catalytic activity of halohydrin dehalogenases towards spiroepoxides

Maja Majerić Elenkov,*a Ines Primožič,b Tomica Hrenar,b Ana Smolko,a Irena Dokli,a Branka Salopek Sondi,a Lixia Tangc

a Ruđer Bošković Institute, Bijenička c. 54, 10000 Zagreb, Croatia. Fax: ++385 1 4680-108; Tel: +385 1 4560965; E-mail: [email protected] b Faculty of Science, University of Zagreb, Zagreb, Croatia. Fax: +385 1 4606 401; Tel: +385 1 4606 400; E-mail: [email protected] c University of Electronic Science and Technology, No.4, Section 2, North Jianshe Road, Chengdu, China. Fax: +86 28 83208238; Tel: +86 28 83208232; E-mail: [email protected]

Content. 1. GC analyses S2 2. Reaction profiles for the non-enzyme ring opening of spiroepoxides S3 3. Docking studies of 1-oxaspiro[2.5]octane compounds 1b and 1d–1f in the active site of HheA and HheC

3.1. Estimated lowest Gibbs energies of binding S5 3.2. Overlay of equatorial and axial lowest energy conformers in the active site of HheA for 1d and 1f enantiomers S6 3.3. Overlay of equatorial and axial lowest energy conformers in the active site of HheC for 1d and 1f enantiomers S8

4. NMR spectra S10

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2012

S2

1. GC analyses

GC analyses for compounds were carried with β-DEX 225 column. Details are given in Table S1. Table S1. GC analyses

Conditions Retention time (min)

Epoxide Azido alcohols 100 °C 2 min, 10 °C/min to 150 °C, 10 min at 180 °C 3.7 (1a) 9.9 (2a) 9.6 (2a') 110 °C 2 min, 10 °C/min to 180 °C, 10 min at 180 °C 4.4 (1b) 10.2 (2b) 10.7 (2b') 120 °C 2 min, 10 °C/min to 200 °C, 10 min at 200 °C 5.2 (1c) 10.8 (2c) 11.4 (2c') 110 °C 5 min, 15 °C/min to 150 °C, 10 min at 150 °C 5.0 (1d) 13.1 (2d) 110 °C 5 min, 15 °C/min to 150 °C, 10 min at 150 °C 4.5 (1e) 13.1, 13.3 (2e) 110 °C 5 min, 15 °C/min to 150 °C, 10 min at 150 °C 5.4 (1f) 13.8 (2f)

Determination of enantiomeric excesses was performed using different chiral columns, Table S2. Table S2. Chiral GC analyses

Comp. Column Conditions Retention time (min)

1d β-DEX 225 80 °C 11.3 (3R,4S)/11.5 (3S,4R)

2d Lipodex E 60 °C 0 min, 2 °C/min to 150 °C 37.2 (3S,4R)/37.6(3R,4S)

1e CP Chirasil DEX 130 °C 5.1 (3R,5R)/5.3 (3S,5S)

2e β-DEX 225 110 °C 5 min, 15 °C/min to 150 °C, 10 min at 150 °C

13.1 (3R,5R)/13.3 (3S,5S)

1f Chiraldex G-TA 60 °C 32.9 (S)/35.8 (R)

2f CP Chirasil DEX 140 °C 0 min, 5 °C/min to 170 °C 9.3(S)/9.5 (R)


S3

2. Reaction profiles for the non-enzyme ring opening of spiroepoxides

Figure S1. Calculated reaction profile for nucleophilic attack of azide on 1a. R stands for reactants, TSα is the transition state for the attack on the Cα and TSβ for the attack on the Cβ carbon atom. Pα and Pβ are products, respectively. Calculations were carried out at the B3LYP/6-311++g(d,p) level with the polarizable continuum model using the integral equation formalism variant in H2O (ε = 78.3553). Gibbs energies were calculated at 298.15 K and 1 atm.

Figure S2. Calculated reaction profile for nucleophilic attack of azide on 1b. R stands for reactants, TSα is the transition state for the attack on the Cα and TSβ for the attack on the Cβ carbon atom. Pα and Pβ are products, respectively. Calculations were carried out at the B3LYP/6-311++g(d,p) level with the polarizable continuum model using the integral equation formalism variant in H2O (ε = 78.3553). Gibbs energies were calculated at 298.15 K and 1 atm.


S4

Figure S3. Calculated reaction profile for nucleophilic attack of azide on 1c. R stands for reactants, TSα is the transition state for the attack on the Cα and TSβ for the attack on the Cβ carbon atom. Pα and Pβ are products, respectively. Calculations were carried out at the B3LYP/6-311++g(d,p) level with the polarizable continuum model using the integral equation formalism variant in H2O (ε = 78.3553). Gibbs energies were calculated at 298.15 K and 1 atm.


S5

3. Docking studies of 1-oxaspiro[2.5]octane compounds 1b and 1d–1f in the active site of HheA and HheC

3.1. Estimated lowest Gibbs energies of binding

Table S3. Estimated lowest Gibbs energy of binding for distinct conformational clusters of ligands (enzyme

1ZMO-HheA, rms tolerance for clustering was 0.5 Å) by the docking studies with the AutoDock 4.2 suite of

programs. O-ax and O-eq stands for the O-axial and O-equatorial conformer respectively.

Comp. (3R)-enantiomers 1kcal mol

G

Contribution in

cluster / %Comp.

(3S)-enantiomers 1kcal mol

G

Contribution in

cluster / %

1a −4.71 100

1bO-ax −5.39 100

1bO-eq −5.36 100

1c −6.06 100

(3R,4S)-1dO-ax −5.78 100 (3S,4R)-1dO-ax −5.63 −5.58

97 3

(3R,4S)-1dO-eq −5.70 100 (3S,4R)-1dO-eq −5.75 100

(3R,5R)-1eO-ax −5.90 100 (3S,5S)-1eO-ax −5.71 −5.70

1 99

(3R)-1fO-ax −6.38 100 (3S)-1fO-ax −6.04 100

(3R)-1fO-eq −5.87 100 (3S)-1fO-eq −6.12 100

Table S4. Estimated lowest Gibbs energy of binding for distinct conformational clusters of ligands (enzyme

1ZMT-HheC, rms tolerance for clustering was 0.5 Å) by the docking studies with the AutoDock 4.2 suite of

programs. O-ax and O-eq stands for the O-axial and O-equatorial conformer respectively.

Comp. (3R)-enantiomers 1kcal mol

G

Contribution in

cluster / %Comp.

(3S)-enantiomers 1kcal mol

G

Contribution in

cluster / %

1a −5.44 100

1bO-ax −5.90 100

1bO-eq −5.65 100

1c −5.64 100

(3R,4S)-1dO-ax −5.48 100 (3S,4R)-1dO-ax −5.62 100

(3R,4S)-1dO-eq −5.68 100 (3S,4R)-1dO-eq −5.54 100

(3R,5R)-1eO-ax −6.61 100 (3S,5S)-1eO-ax −5.69 100

(3R)-1fO-ax −6.78 100 (3S)-1fO-ax −5.81 100

(3R)-1fO-eq −5.95 100 (3S)-1fO-eq −6.36 100


S6

3.2. Overlay of equatorial and axial lowest energy conformers in the active site of HheA for 1d and 1f enantiomers

Fig. S4 (3R,4S)-1d-HheA complexes derived from docking studies: overlay of (3R,4S)-1dO-ax (grey) and

(3R,4S)-1dO-eq (white). HheA active site is represented by some structurally important amino acids. Only polar hydrogen atoms are presented. Interatomic distances are given in Å.

Fig. S5 (3S,4R)-1d-HheA complexes derived from docking studies: overlay of (3S,4R)-1dO-ax (grey) and (3S,4R)-1dO-eq (white). HheA active site is represented by some structurally important amino acids. Only polar

hydrogen atoms are presented. Interatomic distances are given in Å.


S7

Fig. S6 (R)-1f-HheA complexes derived from docking studies: overlay of (R)-1fO-ax (grey) and (R)-1fO-eq (white). HheA active site is represented by some structurally important amino acids. Only polar hydrogen atoms are

presented. Interatomic distances are given in Å.

Fig. S7 (S)-1f-HheA complexes derived from docking studies: overlay of (S)-1fO-ax (grey) and (S)-1fO-eq (white).

HheA active site is represented by some structurally important amino acids. Only polar hydrogen atoms are presented. Interatomic distances are given in Å.


S8

3.3. Overlay of equatorial and axial lowest energy conformers in the active site of HheC for 1d and 1f enantiomers

Fig. S8 (3R,4S)-1d-HheC complexes derived from docking studies: overlay of (3R,4S)-1dO-ax (grey) and

(3R,4S)-1dO-eq (white). HheC active site is represented by some structurally important amino acids. Only polar hydrogen atoms are presented. Interatomic distances are given in Å.

Fig. S9 (3S,4R)-1d-HheC complexes derived from docking studies: overlay of (3S,4R)-1dO-ax (grey) and (3S,4R)-1dO-eq (white). HheC active site is represented by some structurally important amino acids. Only polar

hydrogen atoms are presented. Interatomic distances are given in Å.


S9

Fig. S10 (R)-1f-HheC complexes derived from docking studies: overlay of (R)-1fO-ax (grey) and (R)-1fO-eq (white). HheC active site is represented by some structurally important amino acids. Only polar hydrogen atoms

are presented. Interatomic distances are given in Å.

Fig. S11 (S)-1f-HheC complexes derived from docking studies: overlay of (S)-1fO-ax (grey) and (S)-1fO-eq (white). HheC active site is represented by some structurally important amino acids. Only polar hydrogen atoms are

presented. Interatomic distances are given in Å.


S10

4. NMR spectra

SpinWorks 2.4: 1-Oxaspiro(2,4)heptane

PPM 6.8 6.4 6.0 5.6 5.2 4.8 4.4 4.0 3.6 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4

0

.999

2

.407

2

.425

2

.839

1

1

.959

7

1.9

558

1

.932

4

1.9

257

1

.912

9

1.8

884

1

.884

6

1.8

759

1

.873

9

1.8

666

1

.849

7

1.8

431

1

.832

2

1.8

267

1

.818

9

1.8

173

1

.793

3

1.7

877

1

.779

5

1.7

781

1

.762

2

1.7

567

1

.755

0

1.7

529

1

.751

4

1.7

469

1

.739

6

1.7

191

1

.715

2

1.6

956

1

.694

2

1.6

914

1

.685

3

1.6

786

1

.669

3

1.6

591

1

.655

9

1.6

540

1

.650

9

1.6

406

1

.639

5

1.6

375

1

.610

4

1.6

094

1

.605

7

file: C:\Documents and Settings\Catbio\My Documents\MAJA\NMR\1-Dnevnik br. 13\49-13-1H èisti epox\fid expt: <zg30>transmitter freq.: 300.132701 MHztime domain size: 32768 pointswidth: 6172.84 Hz = 20.567034 ppm = 0.188380 Hz/ptnumber of scans: 24

freq. of 0 ppm: 300.130003 MHzprocessed size: 32768 complex pointsLB: 0.000 GB: 0.0000

SpinWorks 2.5: 1-Oxaspiro[2.4]heptane

PPM 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0

77

.21

97

77

.00

83

76

.79

68

65

.20

97

52

.14

42

32

.46

70

25

.25

79

file: C:\Documents and Settings\catbio\Desktop\E6-7\3\fid expt: <zgpg30>transmitter freq.: 150.917899 MHztime domain size: 65536 pointswidth: 35971.22 Hz = 238.349615 ppm = 0.548877 Hz/ptnumber of scans: 405


Et2O Et2O

O

O


S11

SpinWorks 2.5: CATBIO I.Stipetic IS-E-6-5 APT

PPM 80.0 76.0 72.0 68.0 64.0 60.0 56.0 52.0 48.0 44.0 40.0 36.0 32.0 28.0 24.0 20.0 16.0

77

.41

92 7

6.9

961

76

.57

28

54

.45

92

53

.38

09

33

.66

49

25

.26

95 2

4.8

932

file: C:\Dokumenti\Irena\Epoksidi\E-6-5-dest\IS-E-6-5-apt\1\fid expt: <jmod>transmitter freq.: 75.475295 MHztime domain size: 65536 pointswidth: 17985.61 Hz = 238.297995 ppm = 0.274439 Hz/ptnumber of scans: 4251


SpinWorks 2.5: 1-Oxaspiro[2.5]octane

PPM 7.6 7.2 6.8 6.4 6.0 5.6 5.2 4.8 4.4 4.0 3.6 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4 -0.0 -0.4

1.

000

1.

055

4.

136

7.

263

0

5.

297

4

2.

587

0

1.

750

4

1.7

492

1.

744

5

1.7

290

1.

602

5

1.6

014

1.

597

9

1.5

954

1.

577

3

1.5

598

1.

546

5

1.5

342

1.

509

7

1.4

974

1.

494

9

1.4

941

1.

483

9

1.4

826

1.

481

1

-0

.00

00

file: C:\Dokumenti\Irena\Epoksidi\E-6-5-dest\1\fid expt: <zg30>transmitter freq.: 300.131853 MHztime domain size: 32768 pointswidth: 6172.84 Hz = 20.567092 ppm = 0.188380 Hz/ptnumber of scans: 13


O

O

CH2Cl2


S12

SpinWorks 2.5: 1-Oxaspiro[2.6]nonane

PPM 7.2 6.8 6.4 6.0 5.6 5.2 4.8 4.4 4.0 3.6 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4 -0.0

0

.99

9

6

.27

8

7

.264

0

2

.596

0

1

.709

8

1.6

968

1

.681

4

1.6

321

1

.629

1

1.6

275

1

.613

2

1.6

118

1

.604

4

1.6

035

1

.589

4

1.5

851

1

.559

2

0

.000

1

file: C:\Dokumenti\Irena\Epoksidi\IS-E-6-6\1\fid expt: <zg30>transmitter freq.: 300.131853 MHztime domain size: 32768 pointswidth: 6172.84 Hz = 20.567092 ppm = 0.188380 Hz/ptnumber of scans: 8


SpinWorks 2.5: 1-Oxaspiro[2.6]nonane (3)

PPM 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0

77

.42

15

77

.02

32

76

.99

69

76

.57

43

60

.50

77

55

.68

61

35

.91

66

28

.90

64

24

.60

16

file: C:\Dokumenti\Irena\Epoksidi\IS-E-6-6\2\fid expt: <jmod>transmitter freq.: 75.475295 MHztime domain size: 65536 pointswidth: 17985.61 Hz = 238.297995 ppm = 0.274439 Hz/ptnumber of scans: 1448


O

O


S13

SpinWorks 2.4: (3R,4S),(3S,4R)-1d

PPM 7.6 7.2 6.8 6.4 6.0 5.6 5.2 4.8 4.4 4.0 3.6 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4

0.

999

1.

038

3.

215

10

.36

7

.27

07

2

.75

93

2.7

436

2

.48

88

2.4

730

1

.76

12

1.7

521

1

.74

46

1.7

385

1

.73

17

1.7

233

1

.70

96

1.7

018

1

.69

99

1.6

829

1

.66

97

1.6

516

1

.63

59

1.6

285

1

.62

71

1.6

183

1

.61

46

1.6

134

1

.60

17

1.5

883

1

.58

54

1.5

779

1

.56

94

1.5

443

1

.53

94

1.5

296

1

.52

07

1.5

008

1

.48

90

1.4

787

1

.47

55

1.4

706

1

.46

94

1.4

556

1

.44

86

1.4

412

1

.43

02

1.4

270

1

.42

07

1.4

142

1

.39

03

0.8

415

0

.81

89

file: C:\Documents and Settings\Catbio\My Documents\MAJA\NMR\1-Dnevnik br 12\Epox 13-IVO_1H\fid expt: <zg30>transmitter freq.: 300.132701 MHztime domain size: 32768 pointswidth: 6172.84 Hz = 20.567034 ppm = 0.188380 Hz/ptnumber of scans: 8


SpinWorks 2.4: (3R,4S),(3S,4R)-1d

PPM 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0

77

.441

6

77.0

179

76

.595

3

60

.937

5

52

.589

8

34

.527

4

32

.915

6

32.6

239

24

.686

6

24.0

068

14

.348

4

file: C:\Documents and Settings\Catbio\My Documents\MAJA\NMR\1-Dnevnik br 12\Epox 13-IVO_13C\fid expt: <jmod>transmitter freq.: 75.475295 MHztime domain size: 65536 pointswidth: 17985.61 Hz = 238.297995 ppm = 0.274439 Hz/ptnumber of scans: 1718


O

O


S14

SpinWorks 2.4: (3R,5R),(3S,5S)-1e

PPM 7.2 6.8 6.4 6.0 5.6 5.2 4.8 4.4 4.0 3.6 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4

0.

999

1.

940

1.

773

0.

915

2.

893

4.

740

2

.63

07

2.6

148

2

.60

96

2.5

937

1

.85

49

1.7

898

1

.78

33

1.7

596

1

.75

26

1.7

421

1

.73

46

1.7

288

1

.72

38

1.7

165

1

.70

85

1.6

941

1

.69

13

1.6

793

1

.65

87

1.6

529

1

.64

71

1.6

390

1

.61

91

1.6

068

1

.59

65

1.5

272

1

.48

82

1.4

823

1

.29

63

1.2

896

1

.28

30

1.2

769

1

.27

01

1.2

451

1

.22

23

1.2

158

1

.21

02

1.2

042

1

.03

65

1.0

247

0

.98

80

0.9

706

0

.95

61

0.9

494

0

.92

99

0.9

084

file: C:\Documents and Settings\Catbio\My Documents\MAJA\NMR\1-Dnevnik br 12\54-12_1H\fid expt: <zg30>transmitter freq.: 300.132701 MHztime domain size: 32768 pointswidth: 6172.84 Hz = 20.567034 ppm = 0.188380 Hz/ptnumber of scans: 24


SpinWorks 2.4: (3R,5R),(3S,5S)-1e

PPM 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0

77

.458

8

77.0

327

76

.610

3

58

.487

9

53

.945

1

41

.512

8

33

.781

4

32.6

544

30

.479

7

23

.410

5

22.2

298

file: C:\Documents and Settings\Catbio\My Documents\MAJA\NMR\1-Dnevnik br 12\54-12_13C\fid expt: <jmod>transmitter freq.: 75.475295 MHztime domain size: 65536 pointswidth: 17985.61 Hz = 238.297995 ppm = 0.274439 Hz/ptnumber of scans: 93


O

O


S15

SpinWorks 2.4: (3R,5R)-1e

PPM 4.4 4.0 3.6 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4

0.

999

2.

041

1.

044

2.

920

1.

009

3.

974

1.

874

2

.61

91

2.6

111

2

.60

31

2.5

951

2

.56

84

1

.82

07

1.8

178

1

.81

47

1.8

091

1

.80

75

1.8

017

1

.79

60

1.7

907

1

.78

42

1.7

727

1

.76

55

1.7

627

1

.75

94

1.7

564

1

.75

43

1.7

477

1

.74

61

1.7

389

1

.73

70

1.7

331

1

.72

80

1.7

214

1

.67

78

1.6

702

1

.66

43

1.6

597

1

.65

42

1.6

527

1

.64

91

1.6

463

1

.64

28

1.6

407

1

.63

16

1.6

256

1

.62

22

1.6

201

1

.58

75

1

.50

31

1.4

831

1

.48

09

1.4

615

1

.27

90

1.2

759

1

.27

31

1.2

700

1

.25

27

1.2

502

1

.24

47

1.2

331

1

.22

94

1.2

267

1

.22

48

1

.00

84

1.0

029

0

.98

92

0.9

842

0

.98

25

0.9

665

0

.96

41

0.9

592

0

.95

59

0.9

479

0

.94

08

0.9

386

0

.92

57

0.9

148

file: C:\Documents and Settings\Catbio\Desktop\R,R-1e_1H\fid expt: <zg30>transmitter freq.: 600.133901 MHztime domain size: 32768 pointswidth: 9541.98 Hz = 15.899760 ppm = 0.291198 Hz/ptnumber of scans: 8


SpinWorks 2.4: (3R,5R)-1e

PPM 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0

76

.704

8

76.4

933

76

.281

9

57

.937

5

53

.414

3

41

.026

9

33

.288

2

32.1

658

29

.967

7

22

.913

6

21.6

937

file: C:\Documents and Settings\Catbio\My Documents\MAJA\NMR\1-Dnevnik br 12\R,R-1e_13C\fid expt: <jmod>transmitter freq.: 150.917899 MHztime domain size: 65536 pointswidth: 39370.08 Hz = 260.870838 ppm = 0.600740 Hz/ptnumber of scans: 1116


O

O


S16

SpinWorks 2.4: (R,S)-1f

PPM 6.8 6.4 6.0 5.6 5.2 4.8 4.4 4.0 3.6 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4

1.

000

0.

998

1.

009

1.

197

2.

016

1.

074

3.

161

3.

070

3.

160

2

.59

11

2.5

829

2

.54

84

2.5

403

1

.78

16

1.7

800

1

.77

09

1.7

596

1

.74

84

1.7

383

1

.72

89

1.6

461

1

.63

75

1.6

260

1

.61

54

1.6

041

1

.59

40

1.5

914

1

.58

21

1.5

333

1

.52

54

1.5

114

1

.50

36

1.4

986

1

.49

30

1.4

916

1

.48

34

1.4

708

1

.46

08

1.4

255

1

.40

34

1.3

362

1

.32

70

1.3

160

1

.04

14

1.0

076

0

.97

64

0.9

729

0

.94

33

file: C:\Documents and Settings\Catbio\Desktop\1\fid expt: <zg30>transmitter freq.: 600.133901 MHztime domain size: 32768 pointswidth: 9541.98 Hz = 15.899760 ppm = 0.291198 Hz/ptnumber of scans: 16


SpinWorks 2.4: (R,S)-1f

PPM 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0

77

.230

0

77.0

180

76

.806

4

57

.651

3

53

.876

0

45

.990

5

38

.373

7

33

.165

4

32.7

293

29

.499

9

28.4

558

20

.899

8

file: C:\Documents and Settings\Catbio\Desktop\4\fid expt: <zgpg30>transmitter freq.: 150.917899 MHztime domain size: 65536 pointswidth: 35971.22 Hz = 238.349615 ppm = 0.548877 Hz/ptnumber of scans: 168


O

O


S17

SpinWorks 2.4: 2a

PPM 7.2 6.8 6.4 6.0 5.6 5.2 4.8 4.4 4.0 3.6 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4

1.

000

3.

187

0.

507

0.

934

7

.26

37

3

.38

27

1

.86

14

1.8

556

1

.85

43

1.8

510

1

.84

83

1.8

472

1

.83

99

1.8

356

1

.83

17

1.8

275

1

.82

48

1.8

235

1

.82

05

1.8

184

1

.81

29

1.8

100

1

.80

90

1

.70

55

1.7

040

1

.70

06

1.6

911

1

.68

61

1.6

836

1

.67

91

1.6

662

1

.66

15

1.6

580

1

.65

30

1.6

516

1

.65

01

1.6

463

1

.63

99

1.6

359

1

.63

39

1.6

282

1

.62

42

1.6

183

1

.61

25

1.6

077

1

.60

22

file: C:\Documents and Settings\Catbio\My Documents\MAJA\NMR\1-Dnevnik br 12\28-12A-1H\fid expt: <zg30>transmitter freq.: 600.133901 MHztime domain size: 32768 pointswidth: 9541.98 Hz = 15.899760 ppm = 0.291198 Hz/ptnumber of scans: 12


SpinWorks 2.4: 2a

PPM 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0

81

.633

5

60

.035

1

37

.426

0

23

.501

8

file: C:\Documents and Settings\Catbio\My Documents\MAJA\NMR\1-Dnevnik br 12\28-12A-13C\fid expt: <jmod>transmitter freq.: 150.917899 MHztime domain size: 65536 pointswidth: 35971.22 Hz = 238.349615 ppm = 0.548877 Hz/ptnumber of scans: 1233


OH

N3

OH

N3


S18

SpinWorks 2.4: 2a'

PPM 7.2 6.8 6.4 6.0 5.6 5.2 4.8 4.4 4.0 3.6 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4

1.

000

2.

507

2.

050

7

.26

39

3

.58

66

1

.82

95

1.8

173

1

.81

09

1.8

008

1

.72

01

1.7

074

1

.70

56

1.6

971

file: C:\Documents and Settings\Catbio\My Documents\MAJA\NMR\1-Dnevnik br 12\28-12B-1H\fid expt: <zg30>transmitter freq.: 600.133901 MHztime domain size: 32768 pointswidth: 9541.98 Hz = 15.899760 ppm = 0.291198 Hz/ptnumber of scans: 12


SpinWorks 2.4: Maja 2a*

PPM 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0

73

.91

86

68

.46

87

33

.92

19

24

.24

69



N3

OH

N3

OH


S19

SpinWorks 2.4: 2b

PPM 7.2 6.8 6.4 6.0 5.6 5.2 4.8 4.4 4.0 3.6 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4

1.

000

5.

159

0.

525

7

.26

34

3

.25

65

1

.71

67

1.6

257

1

.58

63

1.5

755

1

.53

52

1.5

178

1

.46

19



SpinWorks 2.4: 2b

PPM 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0

71

.672

9

61

.502

1

35

.045

6

25

.601

5

21

.721

2

file: C:\Documents and Settings\Catbio\My Documents\MAJA\NMR\1-Dnevnik br 12\10-12B-13C\fid expt: <jmod>transmitter freq.: 75.475295 MHztime domain size: 65536 pointswidth: 17985.61 Hz = 238.297995 ppm = 0.274439 Hz/ptnumber of scans: 794


OH

N3

OH

N3


S20

SpinWorks 2.4: 2b'

PPM 7.2 6.8 6.4 6.0 5.6 5.2 4.8 4.4 4.0 3.6 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4

1.

000

5.

976

7

.26

38

3

.55

97

1

.75

13

1.7

248

1

.57

95

1.5

625

1

.55

87

1.5

432

1

.44

45

1.3

998

1

.32

07



SpinWorks 2.4: 2b'

PPM 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0

69

.373

6

64

.739

2

31

.170

4

25

.481

7

21

.853

7



N3

OH

N3

OH


S21

SpinWorks 2.4: 2c

PPM 7.2 6.8 6.4 6.0 5.6 5.2 4.8 4.4 4.0 3.6 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4

1.

000

6.

919

7

.28

05

3

.26

29

1

.74

48

1.6

885

1

.67

20

1.6

624

1

.64

25

1.6

338

1

.60

93

1.6

003

1

.56

28

1.5

496

1

.47

09

1.2

733

0

.01

68



SpinWorks 2.4: 2c

PPM 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0

75

.647

9

61

.745

9

38

.551

7

29

.720

8

22

.241

5



OH

N3

OH

N3


S22

SpinWorks 2.4: 2c'

PPM 7.6 7.2 6.8 6.4 6.0 5.6 5.2 4.8 4.4 4.0 3.6 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4 -0.0

1.

000

7.

442

0.

545

7

.28

08

3

.51

21

3.4

913

1

.83

93

1.8

143

1

.79

73

1.7

853

1

.76

35

1.7

209

1

.62

03

1.5

701

1

.54

95

1.5

245

1

.50

87

1.4

987

1

.48

87

1.2

752

0

.01

89



SpinWorks 2.4: 2c'

PPM 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0

69

.03

62

68

.49

04

34

.47

72

29

.86

82

22

.58

83



N3

OH

N3

OH


S23

SpinWorks 2.4: 2d

PPM 9.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0

1.

000

1.

020

3.

015

11

.24

7

.26

06

3

.42

31

3.3

828

3

.23

21

3.1

918

1

.77

07

1.7

660

1

.76

00

1.7

544

1

.72

25

1.7

151

1

.71

01

1.6

866

1

.68

21

1.6

750

1

.67

03

1.6

639

1

.65

77

1.6

524

1

.64

51

1.6

413

1

.63

30

1.6

260

1

.57

77

1.5

635

1

.55

66

1.5

443

1

.53

25

1.5

205

1

.49

63

1.4

761

1

.47

53

1.4

630

1

.45

38

1.4

434

1

.43

18

1.4

130

1

.40

43

1.3

925

1

.37

64

1.3

688

1

.35

81

1.3

511

1

.33

77

1.3

326

1

.31

35

1.2

955

1

.26

64

1.2

534

1

.24

30

1.2

155

0

.91

29

0.8

909

file: C:\Documents and Settings\Catbio\My Documents\MAJA\NMR\1-Dnevnik br 12\64-12-1H\fid expt: <zg30>transmitter freq.: 300.132701 MHztime domain size: 32768 pointswidth: 6172.84 Hz = 20.567034 ppm = 0.188380 Hz/ptnumber of scans: 9


SpinWorks 2.4: 2d

PPM 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0

76

.708

3

76.4

973

76

.285

6

53

.415

0

41

.026

5

33

.286

8

32.1

662

29

.969

0

29.1

752

22

.913

1

21.6

981

file: C:\Documents and Settings\Catbio\My Documents\MAJA\NMR\1-Dnevnik br 12\46-12-rs-13C\fid expt: <jmod>transmitter freq.: 150.917899 MHztime domain size: 65536 pointswidth: 35971.22 Hz = 238.349615 ppm = 0.548877 Hz/ptnumber of scans: 939


OH

N3

OH

N3


S24

SpinWorks 2.4: 2e

PPM 7.6 7.2 6.8 6.4 6.0 5.6 5.2 4.8 4.4 4.0 3.6 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4

1.0

00

3.

752

0.

501

1.

634

7

.28

77

3

.23

30

1

.81

83

1.8

069

1

.79

70

1.7

895

1

.78

46

1.7

793

1

.76

61

1.7

486

1

.73

68

1.7

079

1

.68

78

1.6

799

1

.67

08

1.6

675

1

.66

07

1.6

519

1

.64

39

1.6

340

1

.62

61

1.6

226

1

.61

45

1.6

069

1

.60

34

1.5

924

1

.27

40

1.2

689

1

.26

15

1.2

527

1

.25

15

1.2

485

1

.23

23

1.2

184

1

.20

32

1.1

951

1

.19

10

1.1

884

1

.17

31

1.1

719

1

.17

04

1.1

607

0

.97

21

0.9

287

0

.90

75

file: C:\Documents and Settings\Catbio\My Documents\MAJA\NMR\1-Dnevnik br 12\56-12-1H\fid expt: <zg30>transmitter freq.: 300.132701 MHztime domain size: 32768 pointswidth: 6172.84 Hz = 20.567034 ppm = 0.188380 Hz/ptnumber of scans: 8


SpinWorks 2.4: 2e

PPM 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0

77

.456

2

77.0

325

76

.609

2

72

.182

1

62

.924

6

43

.205

9

34

.500

9

34.2

536

27

.420

8

22

.413

7

21

.067

1

file: C:\Documents and Settings\Catbio\My Documents\MAJA\NMR\1-Dnevnik br 12\56-12-13C\fid expt: <jmod>transmitter freq.: 75.475295 MHztime domain size: 65536 pointswidth: 17985.61 Hz = 238.297995 ppm = 0.274439 Hz/ptnumber of scans: 469


OH

N3

OH

N3


S25

SpinWorks 2.4: 2f

PPM 7.2 6.8 6.4 6.0 5.6 5.2 4.8 4.4 4.0 3.6 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4

1.

000

1.

957

2.

956

2.

927

3.

105

2.

983

1.

092

0.

887

7

.28

09

3

.22

60

3.2

059

3

.20

22

3.1

821

1

.82

28

1.8

073

1

.80

57

1.8

016

1

.79

75

1.7

958

1

.78

57

1.7

845

1

.77

88

1.7

746

1

.77

32

1.7

576

1

.67

53

1.6

730

1

.66

31

1.6

586

1

.65

26

1.6

502

1

.64

87

1.6

466

1

.64

29

1.6

404

1

.63

13

1

.52

57

1.5

192

1

.51

28

1.5

063

1

.49

63

1.4

893

1

.48

46

1.4

613

1

.44

75

1.4

417

1

.43

92

1

.26

85

1.2

616

1

.24

68

1.2

400

1

.22

48

1.2

178

1

.20

53

1.1

818

1

.15

70

1.1

512

1

.13

53

1.1

297

1

.10

46

0.9

880

0

.92

07

file: C:\Documents and Settings\Catbio\Desktop\1\fid expt: <zg30>transmitter freq.: 600.133901 MHztime domain size: 32768 pointswidth: 9541.98 Hz = 15.899760 ppm = 0.291198 Hz/ptnumber of scans: 16


SpinWorks 2.4: 2f

PPM 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0 0.0

77

.224

4

77.0

120

76

.799

9

72

.622

3

63

.521

9

46

.638

3

39

.079

1

34

.948

0

33.6

075

30

.495

3

27

.154

1

18

.085

6



OH

N3

OH

N3


SUPPLEMENTARY MATERIALS Catalytic activity of halohydrin ... · S6 3.2. Overlay of equatorial and...

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Transcript of SUPPLEMENTARY MATERIALS Catalytic activity of halohydrin ... · S6 3.2. Overlay of equatorial and...