GLUCOSINOLATES AND THEIR HYDROLYSIS
PRODUCTS–
MULTIFUNCTIONAL COMPOUNDS
GLUCOSINOLATE PUBLICATIONS
CANCER
INSECTS
ARABIDOPSIS
BIOFUMIGATION
GLUCOSINOLATES WHAT ARE THEY ?
NATURAL PRODUCTS PRESENT IN ALL CRUCIFEROUS PLANTS
DIVERSE - PROFILES ORGAN SPECIFIC
OH
HO
HO
Lossen
rearrangement
OH
myrosinase
O
OH
S
R
NOSO 3-K
+
O
HO
HO
OH
1
+
OH
2
ESP/Fe2+
S
CN
[ ]n
5
HS
R
RNCS
- +
NOSO 3 K
3
ESP
like protein
/Fe2+
RSCN
6
4
ESP/Fe2+
RCN + S
7
‘ R ‘ GROUP – KEY
BIOSYNTHESIS LARGELY WORKED OUT
GROUPS IN JENA (MAX PLANCK), COPENHAGEN, NORWICH
R GROUP DERIVED FROM AMINO ACIDS VIA CHAIN ELONGATION AND A
SERIES OF OXIDATION STEPS
Rocket
Broccoli
Arabidopsis thaliana
Mustard
INSECT STRATEGIES
Myzus persicae
Brevicoryne brassicae
Plutella xylostella
Pieris brassicae
DEFENCE
INDOLYL GLUCOSINOLATES
A
1
a
B
LOCALISATION OF
pTGG1::GUS EXPRESSION
IN FLOWER STALK OF
ARABIDOPSIS THALIANA
C
‘S’ cells
Brevicoryne brassicae
PURE APHID MYROSINASE
CRUDE PROTEIN EXTRACT
M. PERSICAE
B. BRASSICAE
L1
PURE PLANT MYROSINASE
L2 L3
L4
Artificial Diet 1% sinigrin - Brevicoryne brassicae
Mustard plant (B. nigra) - Brevicoryne brassicae
35
80
alate
apterous
nmoles sinigrin/aphid
30
nmoles sinigrin/aphid
alate
apterous
25
20
15
10
60
40
20
5
0
0
0
2
4
6
8
10
day
12
14
16
18
20
0
2
4
6
8
10
day
12
14
16
18
20
A.BIPUNCTATA
Larvae first instar
% Mortality
100
80
B. brassica reared on Brassica
nigra
60
B. brassica reared on Brassica
oleracae
B. brassica reared on 0%
sinigrin
40
B. brassica reared on 1%
sinigrin
20
A. pisum reared on Vicia faba
0
L1
PARASITIC WASP - APHIDS
Rocket
Broccoli
Arabidopsis thaliana
Mustard
Confirmation of altered volatile profiles
Results:
Col-5
Col-5 BnigALK 1c
1600000
1600000
1400000
1200000
1000000
1000000
800000
800000
600000
600000
400000
200000
400000
200000
0
0
3
13
23
Time (min)
3-butenyl
isothiocyanate
1400000
1200000
4-methylsulfinylbutyl
isothiocyanate
33
3
13
23
Time (min)
33
Diaeretiella rapae behaviour experiments – preliminary
results
Apparatus:
Naive Diaeretiella rapae - Reared on Col-5
Col-5 uninfested n.s.
Col-5 infested
Col-5 (+AOP2) infested
Col-5 (+AOP2)
P<0.05
uninfested
Col-5 infested Col-5 (+AOP2) infested n.s.
80
60
40
20
0
20
40
60
Percentage of D. rapae responding to each odour
80
GLUCOSINOLATES AND CANCERS
1. BROCCOLI – WELL KNOWN TO HAVE ANTICANCER PROPERTIES
2. KEY COMPONENT GLUCORAPHANIN – SULPHORAPHANE
3. OTHER ITCs PROBABLY ALSO IMPORTANT
4. APPEAR TO MODULATE BIOCHEMICAL PATHWAYS ASSOCIATED WITH
CANCER PATWAYS – SUCH AS APOPTOSIS
HUGE AREA OF RESEARCH
SULPHORAPHANE
COOKING ?????? DESTROYS MYROSINASE AND ESP
MYROSINASE MORE ROBUST THAN ESP
FERMENTATION – HUMAN INOCULUM
Molecular Nutrition & Food Research
Volume 56, Issue 12, pages 1906-1916, 27 OCT 2012 DOI: 10.1002/mnfr.201200225
http://onlinelibrary.wiley.com/doi/10.1002/mnfr.201200225/full#mnfr1856-fig-0003
SCREENING PROGRAMME
SELECTION FOR GSL
METABOLISERS
Figure 5. Metabolic fate of glucoraphanin and glucoerucin in E. coli VL8 and E.
casseliflavus CP1 cells.
SOIL MYROSINASES
IMPACT ON BIOFUMIGATION
BACTERIAL ?
BACTERIA HAVE BEEN IDENTIFIED BUT
NOT REALLY CHARACTERISED
PLANT ORIGIN – ROOTS ?
ROOT MYROSINASES
Q BACTERIAL MYROSINASE V ROOT MYROSINASES
WE HAVE IDENTIFIED AND CHARACTERISED AN EXAMPLE OF A BACTERIA
THAT CAN METABOLISE GLUCOSINOLTES
POTENT MYROSINASE
SOME PARALLEL WITH HUMAN GUT BACTERIA THAT DO THE SAME
Allyl isothiocyanate
Lactobacillus agilis
NMR analysis real time
Grown with sinigrin
DON’T FORGET EPITHIONITRILES
ARABIDOPSIS ECOTYPE SCREEN SHOWS ETNs
DOMINANT
ROOTS ?????????
ETNs LESS VOLATILE BUT
MORE STABLE THAN ITCs
Metabolism by bacteria can be very different
Q Do soil microbes metabolise glucosinolates to
Give ITCs ? More work needed in this area
S
CN
O
S
O
CN
CN
CN
CN
CN
OH
S
S
CN
O
CN
O
CN
CN
CN
CN
OH
1.
2.
3.
4.
5.
IDENTIFICATION OF BACTERIAL MYROSINASE
GENOME DETERMINATION
METAGENOMIC APPROACHES
EVERYONE IS DIFFERENT
METABOLISM STUDIES
INSECT WORK
TOM POPE
ELEANNA KAZANA
LAURIENNE TIBBLES
MATTHEW BRIDGES
ALEX JONES
GLEN POWELL
ROSEMARY COLE (WELLESBOURNE)
ATLE BONES (TRONDHEIM)
PLANT WORK
RALPH KISSEN
MURRAY GRANT AND MARTA DE
TORRES ZABALA (UNIVERSITY OF
EXETER)
ROSLINDA SAJARI
PETER KELLY
BACTERIAL WORK
VIN LUANG-IN
ABDUL ALBASER
AJAN NARBAD AND RICHARD
MITHEN (IFR)
FUNDING BBSRC