FORMULATING DAIRY BASED PRODUCT USING PRO-PREBIOTIC INGRIDIENTS

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1 Food Review Seminar: Update on Dairy Industry, Bogor June 2012 FORMULATING DAIRY BASED PRODUCT USING PRO-PREBIOTIC INGRIDIENTS Ir. Lilis Nuraida, MSc. Ph.D. SEAFAST Center and Departemen Teknologi Pangan dan Gizi Institut Pertanian Bogor Outline Definitions of Pro-Pre Pre and Syn-biotic Application of Probiotic in Dairy Industry Pro- and Pre-biotic in Fermented Dairy Products Pro- and Pre-biotic in non fermented product Microencapsulation to improve viability of probiotic 1

2 Probiotic, Prebiotic and Synbiotic Term Probiotic = for life (Greek) Prebiotic Synbiotic Definition Live microorganisms administered in adequate amounts which h confer a l health benefits to the host FAO/WHO (2001) Nondigestible food ingredient that beneficially affects the host by selectively stimulating the growth and/or activity it of one or a limited it number of bacteria in the colon, and thus improves host health A mixture of probiotic and prebiotic Probiotic foods Probiotic Market The largest share of the market, estimated $13.8 billion in 2008, should reach $17.0 billion in 2013, 4.2 percent growth. Mostly in dairy products, with yogurts, kefir and cultured drinks representing the major categories of probiotic foods. Yogurt products accounted for the largest share of sales, representing percent. Emerging food applications: probiotic cheese, nutrition bars, breakfast cereal, and infant formula. Probiotic supplements. The second largest share, $1.2 billion in 2007, expected to reach $1.7 in 2013, 5.8 percent growth. Capsules, tablets and powders, with capsule the largest share of sales, representing 75 percent. Probiotic ingredients Worth $797.6 million in 2008 and increase to $917 million by the end of 2013, 2.8 percent growth. Probiotics of the lactobacillus genus accounted for the largest share, representing 61.9 percent of total sales in

3 Prebiotic market Projected to reach nearly $1.2 billion and $225 million, respectively, by the year 2015, in the European and the U.S. market (Global Industry Analysts). Rapidly rising in popularity within the functional food market: Applications in dairy products, health drinks, nutrition bars, breakfast cereals, beverages, bakery products, meat products, mineral supplements, weight loss products, green foods, infant food and pet food. Microflora Probiotic bacteria Species Lactobacilli Bifidobacteria Enterococci Lactococci Lactobacillus acidophilus L. rhamnosus L. reuteri L. casei L. gasseri L. plantarum L. jhonsonii Bifidobacterium bifidum B. longum B. Breve B. infantis B. adolescentis Enterococcus faecalis E. faecium Lactococcus lactis subsp lactis Not all species/strains are equal Probiotic properties are strain dependent Each strains has to be characterised 3

4 Viability of probiotic bacteria Viability, physiological and metabolic activity of probiotic bacteria in a food product at the point of sale are important consideration for their efficacy They have to survive during shelf life of a food, transit through high acidic and alkaline conditions in the gastro-intestinal tract Probiotic bacteria should be present in a food to minimum concentration of 10 6 cfu/g or the daily intake should be about 10 9 cfu/g Fermented Milk and Lactic Acida Bacteria Beverages Association, Japan: minimum 10 7 cfu/ml to be present in dairy products National Yoghurt Association, USA specifies 10 8 cfu/g at the time manufacture Prebiotic effect of various oligosaccahrides Carbohydrate Nondigestibility Fermentation Selectivity Prebiotic status Inulin and oligofructose Yes Yes Yes Yes Galactooligosacc Probable? Yes Yes harides Lactulose Probable? Yes Yes Isomaltoligosacc Partly Yes Promising No harides Lactosucrose NA NA Promising No Xylooligosacchrai des NA NA Promising No Soybean oligosaccharides NA NA NA No Glucooligosaccha rides NA NA NA No Roberfroid,

5 Use of Prebiotics in food Safety of ingredient is a must and good sensory properties desirable Good prebiotics are stable under heat and when dried, can be stored at room T for months A daily dose of 5-8g/d FOS or GOS has a prebiotic effect in adults Doses higher than 20 g/day might induce some side effects, such as increased flatulence or abdominal bloating. Application of Probiotic in Dairy Industry Fermented products: Yoghurt and fermented milk drink Cheeses Non-fermented products: Ice cream and milk based dessert Powdered milk for infant Butter, Mayonnaise, Fat spread 5

6 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 probiotic microorganism to grow in the medium to increase the cell number The robustness of microoganism to withstand the freezing or drying of starter culture Several aspect must be considered during the production of probiotic fermented milk drink Many probiotic strain grow slowly in non-supplemented milk due to limited proteolytic activity Supplemented with hydrolised d protein, whey derivatives, or amino acid The production condition are often unsuitable for their growth Optimum temperature for probiotic isolated from human is 37 o C Conventional yoghurt fermentation done at 42 o C The metabolites of probiotics may be undesirable due to formation off flavor Bifidobacteria produce acetic acid and lactic acid in the proportion 3:2 which give vinegar like taste 6

7 Yoghurt and fermented milk drink Conventional yoghurt is fermented by L. bulgaricus subsp delbruekii and S. thermophilus not very resistant to the bile released into the small intestine not maintained alive in the gastrointestinal tract in very high numbers Bio yoghurt is yoghurt contain live probiotic microoganisme, i.e. Lactobacillus acidophillus and strains of Bifidobacterium the presence of which may provide beneficial health effect (Laurent Hattingh and Viljoen, 2001). Incorporation of probiotic into fermented milk Add the probiotic microorganisms together with the starter culture (as Direct to Vat Innoculation/DVI 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 culture. The two batches are then mixed together Probiotic microorganism(s) may be used as starter culture, the fermentation may be longer 7

8 Yoghurt fermentation Homogenized milk TS 12% Prebiotic Incubation at 43.3, to reach ph 4.8 Heating and cooling Cooling and Holding Cooling, agitating, Addition starter culture Set yoghurt Packaged in small container Or cup Packed in cups Bulk container Probiotic Stirred yoghurt Factors affecting L. 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 H 2 O 2 produced by L. bulgaricus is detrimental to L. acidophillus Synergistic growth between L. acidophillus and Bifidobacterium 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 8

9 Improvement of survival L. acidophillus and Bifdobacteria in bio-yoghurt...1 Prevention over acidification: Applying heat shock before addition of probiotic culture Lowering storage temperatur to 3-4 o C Improving buffering capacity by the addition of whey protein concentrate Modification of incubation temperature Incubation temperature of 37 o C favours the growth of bifidobacteria Rate of innoculation Some probiotic bacteria grow poorly in milk use a large innoculum 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 may be detrimental t to probiotic culture adding probiotic bacteria after fermentation or adding yoghurt starter culture at later stage Addition of growth promoting substances Supplementation of casitone, casein hydrolysate, fructose, whey protein concentrate improved viability of L. acidophillus Cystein, acid hydrolysate and tryptone improved viability of bifidobacteria Microencapsulation Type of packaging container Bifidobacteria is anaerobic, while lactobacilli is microaerophilic 9

10 Effect prebiotic on bio-yoghurt Addition of oligosaccharides potentially enhance viability bifidobacteria: Use of lactulose in fermented milk improved quality of fermented skim milk by L. acidophilus, L. rhamnosus, Lactobacillus bulgaricus and Bifidobacterium lactis in coculture with Streptococcus thermophilus. Increased the counts of all probiotics, with particular concern to B. lactis (bifidogenic effect) Inulin addition to co-cultures and cocktail: Enhanced products firmness, Increase in microbial growth induced by metabolic interactions among lactic acid bacteria and partial inulin metabolization. (Oliviera et al., 2011) Probiotic Cheeses Natural cheese has proven to be a good carrier for probiotic cultures. Studies have suggested that consuming probiotics in a cheese matrix is favorable for the viability of probiotics through 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 of cheese Should not interfere with the normal activity of other essential microorganisms in the cheese Should be compatible and not produce antimicrobial compounds Should be able to grow on starter culture media 10

11 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 starter cultures 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 hard cheese Addition of freeze dried culture to mtrix of Cheddar cheese following cheddaring and salting Addition of fermented cream dressing in cotage cheese Cream dressing is added for flavour and texture development Two stage fermentation: Fermentation with probiotic bacteria for 2 h followed by fermentation with starter culture The lactic starter culture grow faster than probiotic bacteria Standardized milk Whey draining Probiotic Starter culture Cheddaring Rennet Milling Curd Cutting Probiotic Salting Cooking Hooping and Pressing The manufacturing stage of Cheddar Cheese (Tamime et al., 2007) Lilis Nuraida Juni

12 Ice cream and frozen milk based dessert The ice cream matrix might be a good vehicle for probiotic culture due to its composition (milk protein, fat, lactose and other compounds) During manufacturing ice cream, freezing involves vigorously agitating to incorporate air Detrimental to probiotic bacteria: Lactobacilli are microaerophilic, bifidobacteria are anaerobic Freeze stress must be considered with respect to viability during manufacture and extended storageimpact of probiotic bacteria on flavor should be considered as ice cream 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 bacteria prior to blending with ice cream mix frozen yoghurt ice cream Protection of the probiotic cells againts freeze damage is important (use cryoprotectant) Freeze dried/ Spray Dried encapsulated probiotic Prebiotic Ice cream mix Blend, Freeze and Stored Natural yoghurt containing gprobiotic Pro or synbiotic ice cream Fermented frozen dessert 12

13 Problems during processing ice-cream containing probiotic cultures Step Problem Solutions Formulation with fruit High acidity of the final product may lead to decreased probiotic survival Preferably, use of fruit pulp/juices with a lower natural acidity pulp/juice Some ingredients may have inhibitory Check k for inhibitory activity it of the activity against probiotic strains food ingredients to be used against the probiotic strains to be employed Fermentation Probiotic bacteria show some loss in viability at low ph values Control of the ph during the fermentative process Increased inoculum concentration Selection of strains tolerant to low ph values Beating Storage Oxygen represents a factor of toxicity for probiotic bacteria Stress induced by freezing reduces the viability of probiotic bacteria by at least 1 log cycle Cruz et al., 2009 Selection of oxygen-tolerant strains Increased inoculum concentration Avoid temperature oscillations during storage of the product Stability of the probiotic cultures during ice-cream storage Probiotic cultures wee capable maintaining their stability in frozen food products 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- cream containing i oligofructose Improvement stability: the use of microencapsulation of cultures and the supplementation with prebiotics Cruz et al., (2009) 13

14 Application of probiotic in powdered/dried dairy products Liquid materials + liquid ingredient Spray drying Dried encapsulated probiotic Dry base Mixing Powdered dairy product Dry ingredient Microencapsulation Technique to improved viability Microencapsulation: a process whereby the cells are retained within the encapsulating membrane Improving viability, survival during freezing, stability during storage Entrapment in gelatin, calcium alginat, xanthan-gellan or vegetable gums Encapsulated cells can be dried d to produce cell powder/granule (freeze drying, spray drying, fluidized bed drying) Protection againts adverse conditions in the product Protection againts deleterious conditions in the upper GI tract 14

15 Technique for microencapsulation Atomization: Spray drying Spray Chilling Extrusion Technique Emulsion Technique Freeze Drying Technique for microencapsulation Atomization Spray drying is a commonly used method of encapsulation in the food industry: controlled by means of the product feed,gas flow and temperature Carrier: polysaccharides, lactose, proteins, skim milk, gelatin, soluble starch and gum arabic Skim milk has proved to be a better wall material than gelatin, soluble starch and gum arabic Spray Chilling: the atomization step is similar to spray drying, but the solidifcation of gel particles is based on the injection of cold air into the vessel Freeze drying Cryoprotectan: fructose, lactose, mannose, monosodium glutamate, sorbitol, trehalose 15

16 Technique for microencapsulation Extrusion Technique adding microrganisms to a hydrocolloid solution, and extruding the cell suspension through a syringe needle to free-fall into a hardening solut 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 phase), homogenized to form a water-in-oil emulsion, cross-linked to form insoluble tiny gel particles within the oil phase Supporting materials: K-karagenan, locust bean gum, cellulose actetate pthalate, alginate, chitosan and gelatin Extrusion Technique Emulsion Technique Krasaekoopt et al., 2003 Lilis Nuraida Juni

17 Example of microencapsulated probiotics application in Dairy Products Burgain et al Example of microencapsulated probiotics application in Dairy Products Lilis Nuraida Burgain June et al

18 Thank you ; 18