Development of Vegetables with Improved Health-Promoting Qualities

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Development of Vegetables with Improved
Health-Promoting Qualities

• Elizabeth Jeffery
• Department of Food Science and Human Nutrition,
  University of Illinois

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Fruits, Vegetables Cancer Prevention
Epidemiology is undecided
Variety or Category

Positive
Vegetables
80 (59/74)
Fruits
64 (36/56)
Raw vegetables
87 (40/46)
Cruciferous
Vegetables
69 (38/55)
Allium
Vegetables
77 (27/35)
Green vegetables
77 (68/88)
Carrots
81 (59/73)
Tomatoes
71 (36/51)
Citrus Fruit
66 (27/41)
The 1997 World Cancer Research Fund and the
American Institute for Cancer Research
(WCRF/AICR) report Food, Nutrition and the
Prevention of Cancer a global perspective, p442.
by John D Potter and other panel members
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Epidemiology of Dietary Cancer Prevention Fruit
and Vegetables
Case-Control Studies
Cohort Studies
Riboli and Norat, 2003
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Food Variability environment, genotype
Human Variability environment, genotype
SAFE EFFECTIVE DOSE
BFC i.d. Formulation/ food preparation
Bioavailability Biomarkers
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Human Variability
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Human Variability
Crucifers Lower Risk for Lung Cancer More
Effectively in those at high risk
Relative risk
Crucifers/ non-smokers 0.70 (ns)
Crucifers/smokers 0.31 (plt0.05)
Zhao et al, 2001, Cancer Epi Bio Prev 101063-7
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Breast Cancer Risk, Dietary Crucifers and the
GSTT1 Null Phenotype
Human Variability
OR, Breast Cancer
Quartile of Crucifer Intake
Fowke et al., Cancer Res. 63 3980-3986, 2003
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2005 and 2003 USDA Nutritional Data for RAW
BROCCOLI (abridged) Mean value per 100.00 grams
edible part Name Unit
Amount Amount data S.E.
2003 2005 points Food energy
kcal 28.00 28.00 1 Protein
g 2.98 2.98 22 .11 Total lipid (fat)
g 0.35 0.35 22 .03 Carbohydrate g
5.24 5.24 1 Total saturated fat g
0.05 0.05 1 Cholesterol mg
0 0 1 Total dietary fiber g
3.0 - Vitamin A IU 1542
3000 1 Ascorbic acid mg
93.2 93.2 15 2
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Carotenoid content of Broccoli
Lutein Beta-carotene
10-fold difference
Means, 22 different broccoli genotypes (mmol/100g
DW)
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Glucosinolates precursors to active components
in cruciferous vegetables
glucoraphanin
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Glucosinolates break down to ITC when plants are
crushed or chewed
Glucoraphanin
Myrosinase

ITC Isothiocyanate Anticarcinogen
Sulforaphane
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• VARIATION IN CONTENT
• Determination of scientific basis for
  variation
• CONTROL
• Examples (taken from broccoli)
• Carotenoids and Tocopherols
• Glucosinolates
• Sulforaphane production from glucoraphanin

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Genotype
Genome
Metabolome
Content of metabolite of interest Phenotype
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Genotype
Genome
Environment
Metabolome
Content of metabolite of interest
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Genotype
Genome
Environment
Metabolome
Processing
Content of metabolite of interest in food product
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Glucosinolate variation among 50 broccoli
varieties one season
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(No Transcript)
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Variation in glucosinolates due to genotype,
environment and G x E
Indolyl
Aliphatic
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Genetics x Environment Interaction
G3 G4
Metabolite Content
E1 E2
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Selective increase of the potential
anticarcinogen 4-methylsulphinylbutyl
glucosinolate in broccoli. Carcinogenesis. 1998
19(4)605-9 Faulkner K, Mithen R, Williamson
G.John Innes Centre, Norwich Research Park,
UK.The putative anticarcinogenic activity of
Brassica vegetables has been associated with the
presence of certain glucosinolates.
4-Methylsulphinylbutyl isothiocyanate
(sulphoraphane), derived from the corresponding
glucosinolate found in broccoli, has previously
been identified as a potent inducer of the
anticarcinogenic marker enzyme quinone reductase
NADP(H)quinone-acceptor oxidoreductase in
murine hepatoma Hepa 1c1c7 cells. We have
therefore produced a broccoli hybrid with
increased levels of this anticarcinogenic
glucosinolate and tested the ability of extracts
to induce quinone reductase. A 10-fold increase
in the level of 4-methylsulphinylbutyl
glucosinolate was obtained by crossing broccoli
cultivars with selected wild taxa of the Brassica
oleracea (chromosome number, n 9) complex.
Tissue from these hybrids exhibited a gt100-fold
increase in the ability to induce quinone
reductase in Hepa 1c1c7 cells over broccoli
cultivars, due to both an increase in
4-methylsulphinylbutyl glucosinolate content and
increased percentage conversion to
sulphoraphane.

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Sulforaphane formation glucoraphanin hydrolysis
Glucoraphanin
Myrosinase (Crushing)
Unstable Intermediate
Sulforaphane
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Glucoraphanin Hydrolysis
Glucoraphanin
Myrosinase (Crushing)
Unstable Intermediate
85-90
10-15
Sulforaphane
Sulforaphane Nitrile
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Defatted Broccoli Seed (1 kg)
GC FID Detection
GC FID Detection
Water Extract
Methylene Chloride Extract
Preparative HPLC
Refractive Index
UV 254 nm
Sulforaphane (4.8 g)
Sulforaphane Nitrile (3.8 g)
Matusheski et al, 2001
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Induction of QR in Cell Culture1
1Matusheski and Jeffery, 2001
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Hepatic QR1
Experiment 1
Experiment 2



Significantly different from pair fed group
(Students T-test, plt0.05). Mean Standard
Error
1Matusheski and Jeffery, 2001
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Effect of Heating on Broccoli Florets
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ESP may remove S in glucosinolates that have no
receiving alkenyl group
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Recombinant Epithiospecifier protein Effect of
Heating
Dilution
MW
Std
100 10 1
200-
116-
97.4-
66-
45-
31-
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Broccoli ESP Activity and Bioactivity
Bioactivity
ESP Activity
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ESP
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ESP activity (epithionitrile nitrile) correlated
with the sulforaphane nitrile sulforaphane ratio
P lt 0.05
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How can we harness this in processing to optimize
sulforaphane ?
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Microwave heating for different periods
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Processing to optimize sulforaphane formation
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Sulforaphane excretion
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Genotype
Environment
Phenotype
processing
Stable content of metabolite of interest in food
product
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Acknowledgements

• Jack Juvik
• Barbara Klein
• Mosbah Kushad
• Matthew Wallig
• Richard Mithen
• Malcomb Bennett
• Anna Keck

• Allan Brown
• Anne Kurilich
• In-Gyu Park
• Grace Wang
• Kanta Kobira
• Nathan Matusheski
• Ranjan Swarup