Daniela Barile Associate Professor

Food Science & Technology dbarile@ucdavis.edu

Integrating Biology and New 'Omics to Guide

Diet, Health and Agriculture

Twitter: @danibarile Linkedin: daniela barile

Obrigada pela hospitalidade!

Thank you to Dr. Hanada Viotto, Dr. Gigante, Dr. Gonçalves, Dr. Grimaldi, Dr. Riberiro

Marcella Mazzocato Universidade de São Paulo

Camila Robison Universidade do Vale

do Rio dos Sinos

Leticia de Aquino Universidade Federal

Fluminense

Vitor Luiz Melo Universidade Federal

Fluminense

Dr. Neiva de Almeida Universidade Federal da Paraíba

Dr. Carlos Conte Junior Universidade Federal

Fluminense

Dr. Juliana de Moura

Coming soon…

Dr. Maria Cecilia Ribeiro PhD - UNICAMP

Prof. Mirna Gigante

1 year PostDoc

Research Agenda • Evolutionary Biology

• “Omics” for discovering new bioactives

• Engineering to scale up isolation

• Functional studies to validate the activity

5

Challenges to Agriculture Feed an increasing world population

9 billion by 2050

The 21st century must nourish 9 billion people AND improve the environment AND be energetically sustainable AND use less water AND lead to healthier outcomes…

Residual side streams from the food industry and agriculture

High levels of organic matter Cause pollution if released in the

environment May still contain Bioactive molecules Potential for re-utilization

Targets for improving health?

Identify bioactive compounds in food waste

Mitigate chronic health conditions deriving from poor dietary habits

Our friends…bacteria

• ~ 1 kg

• 100 trillion

microorganisms in a human body

• ~100 times more than human cells

• Microbiota is an “organ”!

• Rise in inflammatory, infectious, metabolic diseases

• Dysregulation in gastrointestinal microbiota

• Our research is identifying new sources of selective

ingredients that can ameliorate those conditions

Feeding for Health

The intestinal microbiota supports: 1. Systemic immunity 2. Gut homeostasis

Its imbalance may cause: 3. Infection

4. Allergic disease 5. Inflammatory bowel disease 6. Metabolic disease

12

Insults to the Microbiota

Broad spectrum antibiotics • Reduce beneficial bacteria and alter balance • Allow proliferation of potentially harmful bacteria

– C. difficile – C. albicans – S. aureus

• Emergence of resistant strains

What to do?

How?

14

Infant

Adult

Bioactive components retained and amplified over 120 million years of evolution

.. Looking for

milk bioactives able to modulate the human gut microbiota

Human Milk Oligosaccharides (HMO)

Zivkovic & Barile, Adv. Nutrition 2011

β 1-4

β 1-6

β 1-4 β 1-3

β 1-3

α 1-3

α 2-6

n

Fu

co

sid

ase

Hexosaminidase

Sia

lida

se

Glc

Gal

GlcNAc

Sialic

acid

Fucose

Indigestible sugars: unique chemical linkages

Beta-galactosidase

SELECTIVELY PROMOTE THE GROWTH OF BENEFICIAL BIFIDOBACTERIA

Translation:

From Breastfed Babies… to Everybody Else?

HUMAN MILK NOT AVAILABLE FOR

MASSIVE EXTRACTION!

..bring this research to practice?

1.6

1.4

1.2

1.0

0.8

0.6

0.4

0.2

0.0

x1

06

3530252015105

Colostrum

Milk Day6

Bovine milk oligosaccharides

Retention Time (min)

Inte

nsity

(cps

)

WOULD NEED MASSIVE AMOUNTS !

Are there abundant food sources of these compounds

that are similarly effective? A new target for Agriculture!

90 kg

WHEY

10 kg

CHEESE

100 kg milk

Enzymatic/acid coagulation

Whey still contains about 50%

of milk nutrients Casein + lipids Other proteins, lactose,

mineral salts etc.

Due to the high biochemical oxygen demand (40,000- g/dm3), it cannot be discharged to surface waters or sewage systems

Brazil: >3 million tons/year

Current Whey Utilization Recovery of proteins by ultrafiltration

Whey protein concentrate or whey protein isolates

($$$)

• High lactose • Pollutant • Disposal Costs

Ultrafiltration

w

WHEY

Whey Permeate food

processing stream

ULTRAFILTRATION

New ‘omics: GLYCOMICS

Glycans: Invisible to common spectroscopy Numerous connectivities and branching Need to obtain structure-function information Need High purity samples Need multiple analytical techniques

OO

CH2OH

OH

OH

OH

OO

CH2OH

OH

OH

OH

Chiral CentersConnection sites

Retention time

ISOMERS

Analytical platform

MALDI-TOF MAXIFLEX Bruker Daltonics

http://barilelab.ucdavis.edu

NANO-LC CHIP Q TOF Agilent Technologies

Increased chromatographic performance

ENZYMATIC DIGESTION

Glyco-proteomics & Glyco-Lipidomics strategy Glycomics strategy

Rapid oligosaccharide identification Co

unts

per

seco

nd (i

nten

sity)

Discovery of Complex Fucosylated Oligosaccharides in cheese whey (mimic human milk)

28

Develop new value for food processing streams:

UCD Milk Processing & Research Lab

Bioguided processing Membrane filtration Enzymatic modification Fermentation

Green technologies

Scalable

No requirement for capital investment

Improve dairy sustainability

1 kg of PURE bioactive milk oligosaccharides

in-vitro and in vivo validation of oligosaccharide activity

$ http://www.sigmaaldrich.com/

Whey Oligosaccharides Functions

Increase beneficial bacteria

(in vitro and anima model)

Decrease pathogenic bacteria

(1st human study)

Restore gut barrier

function (animal model)

Immune system modulation

(in vitro)

Proteins

Peptides

Oligosaccharides

Glycolipids

Glycoproteins

More bioactive compounds!

Social Impact

Premature infants (necrotizing enterocolitis)

Children in developing countries (chronic diarrhea and kwashiorkor)

Cancer, IBD, HIV patients Animal Health:

reducing disease incidence and antibiotic use in calves

Summary: future bioactive ingredients

• Some milk components act as PREBIOTICS stimulating the growth of selected bifidobacteria (PROBIOTICS)

• Oligosaccharides discovery in dairy streams open new possibilities to bring these components to market

• Demonstrated potential to abate microbial imbalances and within the infant and adult’s gut

34

Industry Side Streams

Analytical discovery of bioactives

Characterization and Q.C.

Functional testing

scale up purification

Wine pomace Pistachio shells Coffee grounds

Expanding to other food streams

Apple juice fiber

Obrigada!

Disclosure: I am co-founder of a start-up company (Evolve BioSystems)

The Peter J. Shields Endowed Chair in Dairy Food Science