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FORMULATING DAIRY BASED PRODUCT
USING PRO-PREBIOTIC INGRIDIENTS
Ir. Lilis Nuraida, MSc. Ph.D.SEAFAST Center and
Departemen Teknologi Pangan dan GiziInstitut Pertanian Bogor
Outline
- -
Application of Probiotic in Dairy Industry
Pro- and Pre-biotic in Fermented DairyProducts
Pro- and Pre-biotic in non fermentedproduct
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Microencapsulation to improve viability ofprobiotic
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Probiotic, Prebiotic and SynbioticTerm Definition
Probiotic = for life Live microorganisms administered inree a equa e amoun s w c con er a
beneficial health effect to the hostFAO/WHO (2001)
Prebiotic Nondigestible food ingredient thatbeneficially affects the host byselectively stimulating the growth
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an or ac v y o one or a m enumber of bacteria in the colon, andthus improves host health
Synbiotic A mixture of probiotic and prebiotic
Probiotic Market Probiotic foods
The largest share of the market, estimated $13.8 billion in 2008, should reach $17.0billion in 2013, 4.2 percentgrowth.
Mostly in dairy products, with yogurts, kefir and cultured drinks representing the majorcategories of probiotic foods.
ogur pro uc s accoun e or e arges s are o saes, represen ng . percen .
Emerging food applications: probiotic cheese, nutrition bars, breakfast cereal, andinfant formula.
Probiotic supplements.
The second largest share, $1.2 billion in 2007, expected to reach $1.7 in 2013, 5.8percentgrowth.
Capsules, tablets and powders, with capsule the largest share of sales, representing75 percent.
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ro o c ngre en s
Worth $797.6 million in 2008 and increase to $917 million by the end of 2013, 2.8percent growth.
Probiotics of the lactobacillus genus accounted for the largest share, representing 61.9percent of total sales in 2007.
http://www.naturalproductsinsider.com/news/2008/05/global-probiotics-market-worth-196b-by-2013.aspx
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Prebiotic market Projected to reach nearly $1.2 billion and $225
million, respectively, by the year 2015, in theEuropean and the U.S. market (Global IndustryAnalysts).
Rapidly rising in popularity within the functional foodmarket: Applications in dairy products, health drinks, nutrition bars,
breakfast cereals, beverages, bakery products, meat products,
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, , ,food and pet food.
http://www.nutraceuticalsworld.com/contents/view_breaking-news/2010-02-23/report-finds-significant-potential-in-prebiotics-m/
Microflora Species
Lactobacilli Lactobacillus acidophilusL. rhamnosus
L reuteri
Probiotic bacteria
.
L. casei
L. gasseri
L. plantarum
L. jhonsoniiBifidobacteria Bifidobacterium bifidum
B. longum
B. Breve
Not allspecies/strainsare equalProbiotic
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.
B. adolescentis
Enterococci Enterococcus faecalisE. faecium
Lactococci Lactococcus lactis subsp lactis
properties arestrain dependentEach strains hasto becharacterised
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Viability of probiotic bacteria Viability, physiological and metabolic activity of probiotic
bacteria in a food productat the point of sale are importantconsideration for their efficacy
Probiotic bacteria should be present in a food to minimumconcentration of 106 cfu/g or the daily intake should be about 109
cfu/g
Fermented Milk and Lactic Acida Bacteria Beverages7
,high acidic and alkaline conditions in the gastro-intestinal tract
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,products
National Yoghurt Association, USA specifies 108 cfu/g at the timemanufacture
Prebiotic effect of various oligosaccahrides
Carbo hy drate Nond igestibility Fermentation S electivity PrebioticstatusInulinand
oligofructose
Yes Yes Yes Yes
Galactooligosacc
harides
Probable ? Yes Yes
Lactulose Probable ? Yes Yes
Isomaltoligosaccharides
Partly Yes Promising No
Lactosucrose NA NA Promising No
Xylooligosacchrai NA NA Promising No
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Soybean
oligosaccharides
NA NA NA No
Glucooligosaccha
rides
NA NA NA No
Roberfroid,2007
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Use of Prebiotics in food
Safety of ingredient is a must and goodsensory properties desirable
when dried, can be stored at room T formonths
A daily dose of 5-8g/d FOS or GOS has aprebiotic effect in adults
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some side effects, such as increasedflatulence or abdominal bloating.
Application of Probiotic in Dairy Industry
Fermented products:
Cheeses
Non-fermented products:
Ice cream and milk based dessert
Powdered milk for infant
Butter, Mayonnaise, Fat spread
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The choice of probiotic microoganism for
production of starter culture
Posses health beneficial effect to human
Tolerance to acidity and bile salts
The ability of the probioticmicroorganism to grow in the medium toincrease the cell number
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withstand the freezing or drying ofstarter culture
Several aspect must be considered during theproduction of probiotic fermented milk drink
Many probiotic strain grow slowly in non-supplementedmilk due to limited proteolytic activity
uppemene w y ro se pro en, w eyderivatives, or amino acid
The production condition are often unsuitable for their
growth Optimum temperature for probiotic isolated from
human is 37oC
Conventional yoghurt fermentation done at 42oC
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The metabolites of probiotics may be undesirable due toformation off flavor
Bifidobacteria produce acetic acid and lactic acid inthe proportion 3:2 which give vinegar like taste
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Yoghurt and fermented milk drink
ConventionalyoghurtisfermentedbyL.bulgaricus
.
notveryresistanttothebilereleasedinto thesmall
intestinenotmaintainedaliveinthegastrointestinal
tractinveryhighnumbers
Bioyoghurtisyoghurtcontainliveprobiotic
microoganisme,i.e.Lactobacillusacidophillus and
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strainsof
Bifidobacterium the
presence
of
which
may
providebeneficialhealtheffect(LaurentHattinghand
Viljoen, 2001).
Incorporation of probiotic into
fermented milk
Add the probiotic microorganisms together with the
culture) Probiotics do not usually grow markedly during mixed
fermentation The probiotic microorgnisms may be grown in one
batch of milk to achieve a high viable count, another batch of milk is fermented with traditional starter
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cuture.
The two batches are then mixed together
Probiotic microorganism(s) may be used as starterculture, the fermentation may be longer
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Yoghurt fermentation
Homogenized milkTS 12%
Incubation at 43.3, to reachpH 4.8
Prebiotic
Heating andcooling
Addition starterculture
Cooling,agitating,
Packaged in
Cooling andHolding
Set yoghurt
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Packedin cups
Bulkcontainer
Or cup
ProbioticStirred yoghurt
Factors affectingL. acidophillus and
Bifidobacteria in bio-yoghurt Yoghurt acidity
Most strain of bifidobacteria are sensitive to pH below 4.6
The product should be maintain at pH above 4.6
L. acidophillus is more resistant
Strains
The bifidobacteria should be able to grow in milk. Some lack of proteolytic
activity
Co-culture and species interaction
H2O2 produced by L. bulgaricus is detrimental to L. acidophillus
Synergistic growth between L. acidophillus and Bifidobacterium
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Dissolved oxygen
Bifidobacteria is strictly anaerob
Storage condition
Low temperature restrict the growth of Lactobacillus and so over-acidification
Bifidobacteria less tolerant to low temperature storage
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Improvement of survival L. acidophillus and
Bifdobacteria in bio-yoghurt...................1
Prevention over acidification:
Applying heat shock before addition of probiotic culture
-
Improving buffering capacity by the addition of whey proteinconcentrate
Modification of incubation temperature
Incubation temperature of 37oC favours the growth ofbifidobacteria
Rate of innoculation
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Some probiotic bacteria grow poorly in milkuse a largeinnoculum size or concentrated inoculum
Selection of starter culture
Improvement of survival L. acidophillus and
Bifidobacteria in bio-yoghurt...................2
Two-stage fermentation
Acid and hydrogen peroxyde produce by yoghurt starter culture maye e rmena o pro o c cu ure a ng pro o c ac era a er
fermentation or adding yoghurt starter culture at later stage
Addition of growth promoting substances
Supplementation of casitone, casein hydrolysate, fructose, wheyprotein concentrate improved viability ofL. acidophillus
Cystein, acid hydrolysate and tryptone improved viability ofbifidobacteria
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Microencapsulation
Type of packaging container
Bifidobacteria is anaerobic, while lactobacilli is microaerophilic
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Effect prebiotic on bio-yoghurt
Addition of oligosaccharides potentially enhance viabilitybifidobacteria:
Use of lactulose in fermented milk improved quality ofermen e s m m y . ac op us, . r amnosus,Lactobacillus bulgaricus and Bifidobacterium lactis in co-culture with Streptococcus thermophilus.
Increased the counts of all probiotics, with particularconcern to B. lactis (bifidogenic effect)
Inulin addition to co-cultures and cocktail:
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Enhanced products firmness, Increase in microbial growth induced by metabolic
interactions among lactic acid bacteria and partial inulinmetabolization.
(Oliviera et al., 2011)
ProbioticCheeses Natural cheese has proven to be a good carrier for
probiotic cultures.
Studies have suggested that consumingprobiotics in acheese matrix is favorable for the viabilit of robioticsthrough the digestive tract.
Desirable properties of probiotic bacteria to be incorporated to cheese: Must survive the entire shelf-life of cheese.
Must not produce metabolites that are detrimental to the quality ofcheese
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Should not interfere with the normal activity of other essentialmicroorganisms in the cheese
Should be compatible and not produce antimicrobialcompounds
Should be able to grow on starter culture media
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Introduction of probiotic
microorganism into cheeses Introduce as adjunct cultures together with lactic starter cultures
Risk of losing large numbe rof cells to whey or domination of lactic startercultures
Addition of microencapsulated probiotic
Protected the probiotic from degradation duirng teh aging period
Addition of dried probiotic cultures during salting of curd on semi-hard and hardcheese
Addition of freeze dried culture to mtrix of Cheddar cheese following cheddaringand salting
Addition of fermented cream dressing in cotage cheese
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Cream dressing is added for flavour and texture development Two stage fermentation:
Fermentation with probiotic bacteria for 2 h followed by fermentation withstarter culture
The lactic starter culture grow faster than probiotic bacteria
Standardized milk
Starter culture
Whey draining
CheddaringProbiotic
Rennet
Curd Cutting
Milling
SaltingProbiotic
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Cooking Hooping andPressing
The manufacturing stage of Cheddar Cheese (Tamime et al., 2007)
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Ice cream and frozen milk based
dessert
The ice cream matrix might be a good vehicle for probioticculture due to its composition (milk protein, fat, lactose andother compounds)
During manufacturing ice cream, freezing involvesvigorously agitating to incorporate air
Detrimental to probiotic bacteria:
Lactobacilli microaerophilic, bifidobacteria anaerobic
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during manufacture and extended storageImpact ofprobiotic bacteria on flavor should be considered as icecream is not fermented product
Incorporating probiotic into ice
cream
Direct, i.e. blend the ice cream mix and probiotic cells prior to freezing
Involve fermentation of the milk for proliferation of probiotic bacteriapror o en ng w ce cream mx rozen yog ur ce cream
Protection of the probiotic cells againts freeze damage is important(use cryoprotectant)
Freeze driedencapsulated
Ice cream mix Natural yoghurtcontainin robiotic
Prebiotic
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probiotic
Blend, Freeze and Stored
Fermented frozen dessertPro or synbiotic ice cream
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Problems during processing ice-cream
containing probiotic cultures
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Cruz et al., 2009
Stability of the probiotic cultures during
ice-cream storage
Probiotic cultures wee capable maintaining theirstabilit in frozen food roducts
Addition of prebiotic (inulin and oligofructose) :
Higher overrun for the ice-cream mix with inulin
Less changes in melting properties
Firmer during storage
Increase probiotic survival during storage of the ice-
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cream contanng o go ructose
Improvement stability: the use of microencapsulation ofcultures and the supplementation with prebiotics
Cruz et al., (2009)
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Application of probiotic in
powdered/dried dairy products
Liquid materials +liquid ingredient
Spray drying
Dry base
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Powdered dairy product
Mixing Dry ingredient
Driedencapsulated
probiotic
Microencapsulation: a process
Microencapsulation Technique to
improved viability
Protection againts adverseconditions in the product
the encapsulating membrane
Improving viability, survival during
freezing, stability during storage Entrapment in gelatin, calcium
alginat, xanthan-gellan or vegetablegums
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ncapsua e ce s can e re oproduce cell powder/granule (freezedrying, spray drying, fluidized beddrying)
Protection againts deleteriousconditions in the upper GI tract
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Technique for microencapsulation
Atomization:
Spray Chilling
Extrusion Technique
Emulsion Technique
Freeze Drying
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Technique for microencapsulation
Atomization
Spray drying is a commonly used method of encapsulation in thefood industry:
,
Carrier: polysaccharides, lactose, proteins, skim milk, gelatin,soluble starch and gum arabic
Skimmilk has provedto be a better wall material thangelatin,soluble starch and gum arabic
Spray Chilling:
the atomization step is similar to spray drying, but the solidifcation
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o ge par c es s ase on e nec on o co a r n o e vesse
Freeze drying
Cryoprotectan: fructose, lactose, mannose, monosodium glutamate,sorbitol, trehalose
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Technique for microencapsulation
Extrusion Technique
adding microrganisms to a hydrocolloid solution, andextruding the cell suspension through a syringe needle tofree-fall into a hardeningsolut ion or setting bath
Supporting materials: alginate
Emulsion Technique
Adding small volume of the cell-polymer suspension(discontinuous phase to a large volume of a vegetable oil(continuous hase), homo enized to form a water-in-oil
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emulsion, cross-linked to form insoluble tiny gel particleswithin the oil phase
Supporting materials: K-karagenan, locust bean gum,cellulose actetate pthalate, alginate, chitosan and gelatin
Extrusion Technique Emulsion Technique
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Krasaekoopt et al., 2003
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