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INGREDIENTS probiotics

Delivering

probiotic bacteria
in shelf stable foods
By Arup Nag, Food Technologist, and Dr. Shantanu Das, Product Development Manager, Riddet Ins tute, Massey University, New Zealand

robiotics are live microorganisms that, when administered in adequate amounts, confer a health benet on the host (FAO/WHO, 2001). The physiological benets include enhanced immunity, improved gut health (e.g. prevention of intestinal disturbances, balancing of the intestinal microora, and prevention of various types of diarrhoea), prevention of various types of allergies, improved oral health and weight management. However, to achieve a signicant benet of probiotics, we need to consume high concentration of live bacteria (8 9 log CFU/day). Considering consumption of 100g of such food, the food needs to contain at least 6 log CFU/g viable probiotics cells (million per gram). Probiotic bacteria is one of the top functional ingredients today and underpins some of the most successful functional food brands in the world. Reports of several leading food industry analysts including New Nutritional Business, Mintel and Nutra-ingredients identied gut health as one of the top health and wellness platforms for functional foods, and

probiotics is the most important functional ingredient to deliver the gut health benet. In the last decade, some of the most successful functional food brands leveraged the benet of probiotic bacteria. All these information indicate that probiotics is an opportunity. However, as the market is crowded with numerous players, especially with the well-established big players, new entrants will need a niche with no or low level of competition. As the application of probiotics is limited mainly to foods that are subjected to chilled distribution and storage delivering probiotics through shelf stable foods could be an unexploited opportunity. This will help probiotics to reach new consumers, especially in the developing markets, where refrigerated supply chain is inadequate. It will expand the current market for probiotics, which is estimated to be US$20 billion. However, there are currently few credible delivery technologies that are able to deliver probiotic bacteria in shelf-stable foods.

Figure 1: Schematic process ow of ProBioLife technology.

Figure 2: Potential food applications of the ProBioLife ingredient.

FOOD & BEVERAGE ASIA OCTOBER/NOVEMBER 2011

probiotics INGREDIENTS

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This article will discuss a novel delivery technology for probiotic bacteria, which endeavours to bridge this technology gap. This technology is referred as ProBioLife and the ingredient manufactured using this technology is referred as ProBioLife ingredient. This technology has been recently developed in the Riddet Institute, Massey University, New Zealand, and two provisional patents have been led.

ProBioLife technology
The probiotic culture was grown in a specially designed media under optimised conditions. The cells were harvested at a predetermined stage of the growth cycle. After repeated washings (in order to remove the adhered metabolites), they were encapsulated in a specially designed matrix, containing all food grade natural ingredients having GRAS status. Finally, drying of the whole matrix was accomplished under optimised conditions with minimum possible heat stress to the entrapped bacteria. The nal water activity (aw) of the powdered ingredient was between 0.25 and 0.3. The powdered ingredient was packed in double layered LDPE sachets and stored at 25C. The initial set of experiments was carried out using Lactobacillus casei CRL431 obtained from ATCC, Australia. Later this technology was successfully applied to a number of strains of Lactobacillus and Bidobacterium.

Viability
Viability of L. casei CRL431in ProBioLife ingredient was followed over one year at 25oC, using a freeze dried sample (freeze dried culture mixed with whole milk powder) as control. The initial viable bacterial population of L. casei CRL431 were 10.2 and 9.9 log CFU/g in the ProBioLife ingredient and freeze dried sample respectively. After 12 weeks at 25oC, the freeze dried sample had a total live count of 4.2 log CFU/g well below the recommended level of 6 log CFU/g. In the case of ProBioLife ingredient, a small gradual fall in the cell viability was observed throughout the 52 weeks of storage at 25C. After 36 weeks, the viable bacterial population was over 9 log CFU/g (over 1 billion per gram). The nal viable cell count after 52 weeks of storage at 25C was 8.3 log CFU/g (over 100 million per gram).

Application
ProBioLife ingredient can be added to a range of shelf stable dry and intermediate moisture foods, such as dairybased powder beverage, malted milk food, cereal-based powder beverage, breakfast cereal, cereal bar, protein bar and spread. ProBioLife ingredient can be added to the ready-to-drink beverages through dosing cap. Also, this ingredient can be delivered in various formats in the nutraceuticals and supplements area.

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INGREDIENTS probiotics

Figure 3: Stability of L. casei CRL431 cells in powder beverage (), chocolate spread () and cereal bar () when stored at 25C.

Figure 4: Stability of L. casei CRL431 in ProBioLife ingredient () and in freeze dried sample () at 25C. The freeze dried sample consists of freeze dried bacteria mixed with whole milk powder.

Added water temperature 75C 65C 55C

Original cell population (Log CFU/g) 8.4 8.4 8.4

After 5 Min (Log CFU/g) 7.9 8.3 8.5

After 30 Min (Log CFU/g) 7.3 7.9 8.4

Final mix temperature (C) after 30 Min 35 25 25

Table 1: Effect of hot water addition at different temperatures on L. Casei 431 cell viability when encapsulated in the ProBioLife ingredient.

Stability
The ProBioLife ingredient was added to three commercially available shelf stable food products such as cereal bar, chocolate spread and dairy-based powder beverage. The addition levels were 1.0% (w/w). The cereal bars were repacked in LDPE layered aluminium foils and heat sealed. The chocolate spreads were lled into sterile plastic containers with air tight caps keeping minimal head space. Dairy-based powder beverage was dry blended with ProBioLife ingredient and packed in heat sealed aluminium foil. Samples were stored at 25C with no humidity control. Viable cells were enumerated for six months at four weeks interval. The initial lactobacillus counts in cereal bar, chocolate spread and powder beverage were 7.5, 7.6, 8.6 log CFU/g respectively. At the end of the six months storage period at 25C, the viable cell counts decreased slightly to 6.7, 7.4 and 7.6 log CFU/g respectively. The best stability was achieved in chocolate spread probably because of the additional protection obtained from the vegetable oils present in the chocolate spread. However in all the three examples, the loss of viability was not more that one log over six months at 25C and all the three samples had the recommended level of live cells (1 million per gram).

water is a common practice. Most of the products in this category instruct consumers to add warm water (not boiling water) to make the beverage. Therefore, the application trial has simulated a similar drinking condition by directly adding 90ml of hot water at 55C, 65C, and 75C to 10g of the dairy based powder beverage (fortied with ProBioLife ingredient), and enumerated the live cell counts after 5 and 30 minutes. The 30-min time period was assumed to be the maximum time necessary to drink one serving of hot beverage. The enumeration data is presented in Table 1. The result showed that the minimum concentration of live bacteria in the beverage was 7.3 log CFU/g. Therefore it can be seen that even the hot drink is able to deliver recommended levels of live probiotics (1 million per gram).

Conclusion
Understandably a technology that enables probiotics to be used in shelf stable foods will increase the size of the probiotics market significantly by tapping into additional product categories and additional consumers. This technology will make the benets of probiotic bacteria available to millions of consumers in Asia Pacic countries, where a chilled supply chain is often unavailable. As research work is being continued to further improve the technology, winning the Food Industry Innovation Excellence Award 2011 from the New Zealand Institute of Food Science and Technology in June 2011 is very encouraging for the research team. FBA

Hot beverages
An important criterion for successful delivery of live probiotic bacteria in a dairy-based powder beverage is the heat stability of fortied probiotics because mixing with hot

FOOD & BEVERAGE ASIA OCTOBER/NOVEMBER 2011