Investigating the Biosynthetic Pathway of Polyacetylenic Natural Products in Fistulina hepatica and Echinacea purpurea

Graduate School ETD Form 9 Revised 12/07 PURDUE UNIVERSITY GRADUATE SCHOOL Thesis/Dissertation Acceptance This is to certify that the thesis/dissertation prepared By Anthony S.. Rans

Graduate School ETD Form 9

(Revised 12/07)

PURDUE UNIVERSITY

GRADUATE SCHOOL Thesis/Dissertation Acceptance

This is to certify that the thesis/dissertation prepared

By Anthony S Ransdell

Entitled:

Investigating the Biosynthetic Pathway of Polyacetylenic Natural Products in Fistulina

hepatica and Echinacea purpurea

For the degree of Master of Science

Is approved by the final examining committee:

Robert E Minto

Chair Eric C Long

Lei Li

To the best of my knowledge and as understood by the student in the Research Integrity and

Copyright Disclaimer (Graduate School Form 20), this thesis/dissertation adheres to the provisions of

Purdue University’s “Policy on Integrity in Research” and the use of copyrighted material

Approved by Major Professor(s): Robert E Minto

Approved by: Martin J O’Donnell 10/31/2012

Head of the Graduate Program Date

Graduate School Form 20

(Revised 9/10)

PURDUE UNIVERSITY

GRADUATE SCHOOL Research Integrity and Copyright Disclaimer

Title of Thesis/Dissertation:

Investigating the Biosynthetic Pathway of Polyacetylenic Natural Products in Fistulina hepatica and Echinacea purpurea

For the degree of Master of Science

I certify that in the preparation of this thesis, I have observed the provisions of Purdue University Executive Memorandum No C-22, September 6, 1991, Policy on Integrity in Research.*

Further, I certify that this work is free of plagiarism and all materials appearing in this

thesis/dissertation have been properly quoted and attributed

I certify that all copyrighted material incorporated into this thesis/dissertation is in compliance with the United States’ copyright law and that I have received written permission from the copyright owners for

my use of their work, which is beyond the scope of the law I agree to indemnify and save harmless Purdue University from any and all claims that may be asserted or that may arise from any copyright violation

Anthony S Ransdell

Printed Name and Signature of Candidate

10/31/2012

Date (month/day/year)

iii

∆9c,12c

∆9c,12a ∆9c,12a,14c

Figure

∆9c,12a,14c

∆9c,12a,14a

Figure

∆9c,12a,14a,16

∆9c,12a,14a,16

xi

18:1∆9c

∆9c,12a

2

4-­‐C8

H(X)3-­‐4 2-­‐3 2-­‐3

2

ferredoxin-­‐NADP+

H/k D

H/k D H/k D

∆9c,12OH ∆9c,12OH,15c ∆11c,14OH

∆11c,14OH,17c

∆9c,12aepoxy-­‐18:1∆9c

H/kD

H/kD

∆

∆9c,12a(18:2∆9c,12a,14c

∆9c,12a18:2∆9c,12a,14c

∆9c,12c

∆8t,10t,12c

16

17

20

2

6

∆9c,12a,14c

2

3

f

263

∆9c,12a

29

∆9c,12a

∆9c,12c

∆9c,12a,14c

18:2∆9c,12a,14a,16

A

B

A

B

A

B

f

R

18Z

A

B

A

B

47

18:1∆9c,12a ∆9c,12a,14c ∆9c,12a,14c

th

∆9c,12t

A

∆9c,12c

16:2∆9c,12t16:2 ∆9c,12t

16:2∆9c,12t16:2 ∆9c,12t

18:2∆9c,12a,14a,16c18:2∆9c,12a,14a,16t

∆9c,12a

∆9c,12a,14a18:1∆9c,12a,14a,16 ∆9c,12a ∆9c,12a,14c

∆9c,12a,14c

∆9c,12a,14c,16c/t18:1∆9c,12a ∆9c,12a,14c

∆9c,12a,14a

∆9c,12t

∆9c,12a,14a,16c ∆9c,12a,14a,16t

53

14C]

5518:1∆9c,12a,14a

56

(18:1∆9c,12a

58

(18:2∆9c,12c

616

62

+

6

646

TM

66

∆9c,12a

∆9c,12a

70

18:1∆9c,12a

A

B

75

77

18:1∆9c,12a ∆9c,12a,14c

16:2∆9c,12t ∆9c,12t ∆9c,12c

∆9c,12t18:2∆9c,12c

8018:1∆9c,12a

O OH O OH O OH O OH

O OH

Echinacea

Acetylenic Natural Products

O

O

N H O

O N

82

+

Oligo(dT) 12-­‐18

2

2

2

2

2

H 2

2

2

90

4 ) 2 SO 4

4

2

2 2

2

2 2

95

Δ 6

99

104

MLSIFEDSPEYKKRAQTPFTPTKVTLAEVRAVVPKHLHEKNALKATFYVLRDILCAVAVYKLGWLIDPLTTSLVRDYGFNPTLGSIVKWGLWATYWHWQGVILAGWWCLGHEAGHGSLSSAKWYNTVVGYGLHTFILVPYFSWRSSHHAHHKATMSIERDENFVPRTRKDYGLPPEKIAVLNDYHEIFEEAPFYTLARMLFMQGLGWQYYLFTNALGNPMYPPGTNHFSPSSPLFKPHERNGIIASNIGLTIMSSLLWRWTMSVGINMFLKLYFVPYILANHWIVMLTYLHHSDPTVAYYRASHWSFLRGAVSTVDRPLLGWAGRFFLHNVSHDHVAHHLFSSIPFYNQPEVTEAIKKVLKNDYNYDSTNTFRALYRTFSQCCFVEDDGDVVFYKNRDG

A

B

A

B

∆9c,12a

A

B

A

B

A

B

A

B

116

MGAGGRMTTASDDQKNILERVPCSKPPFSLADLKKAIPPHCFERSVIRSSYYVVHDLAVAYVFYYLANTYIPLLPSPFAYLAWPLYWFCQGSILTGVWVIGHECGHHAFSEYQWIDDTVGFVLHSALFTPYFSWKYSHRNHHANPNSLDNDEGYIPKVKSKVTLYSKILNNPPGRVFTLAFRLILGFPLYLFTNVSGKKYERFANHFDPMSPIFTERERVQVLLSDLGLIAVAYAVRKAVLAKGAAWVMCIYGVPVLVVSAFFVLITYLHHTHLSLPHYDSTEWDWIKGALSTIDRDFGFLNRVFHDVTHTHVLHHLISYIPHYHAKEARDAIKPVLG

125