Advancement in research for improving the ambient temperature stability of a range of probiotic strains


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An overview of my research in a Centre of Research Excellence in New Zealand to develop novel technology which successfuly enabled probiotic strains remain stable for longer duration under ambient storage conditions.

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Advancement in research for improving the ambient temperature stability of a range of probiotic strains

  1. 1. Shelf stable probiotic foods –Now a reality Arup Nag29.06.201129.06.2011
  2. 2. Arup Nag29.06.2011Arup Nag29.06.2011AgendaIdentifying the opportunityCore technologyComparison with selected established technologiesPath to commercialization
  3. 3. Health benefits of probiotics are:•Immunity enhancement•Improved gut healthPrevention of intestinal disturbancesBalancing intestinal micro floraPrevention of various types of diarrhoea•Prevention of certain types of allergenic reaction in infants•Improved oral healthEnormous body of scientific research to show the health benefits of probiotic bacteriaMost common genus: Lactobacillus and BifidobacteriumWhat are probiotics?Probiotics are live microorganisms which when administered in adequate amountsconfer a health benefit on the host (FAO/WHO, 2001)
  4. 4. Probiotics marketProbiotics market is approaching close to 20 billion dollars.....further growth possible withtechnology that helps reach new consumers
  5. 5. 20072000YogurtOnlyYogurt, fermented milk, cottage cheese, soyand more00.‘Danonedairy–theaccelerationofgrowth’presentationbyBernardHoursinDanoneinvestorseminar2007Source:MintelGNPDDanone Activia is one of the most successful food product in recent time – this is positioned asdigestive food based on scientifically proven probiotic bacteria - Activia grew over 600% in 7 yearsSuccess of Activia demonstrates the high potential of probiotic as a sustainable and highly crediblefunctional ingredientReview of secondary market data
  6. 6. Hard to compete with the established players within the existing categories ........... hugeopportunity in extending offerings from chilled shelf to ambient shelfTechnology for deliveringprobiotics in shelf stable foodsTechnology gapOpportunity in probiotics areaEnabling technology with sufficient technical insulation to unlock untappedopportunities
  7. 7. Consumption of probiotic bacteria 108 – 109 cfu/g per day for health benefitsConsidering consumption of 100 g probiotic rich food per day, probiotic rich foodneeds to contain 106 – 107 cfu/g viable cells of probiotic bacteriaChallenges in maintaining viability of probiotic cells during processing, storageand gastro-intestinal transitApplication in probiotics restricted to chilled yoghurt, fermented milk and juiceLimited technology available for shelf stable food and beverageChallenges in delivering probioticsProcessing•High temperature•Low water activity, High acidStorage•Time, Temperature•Low water activity , High acid•High acidity in stomach•Bile salt in intestineGI tractHarsh conditions during processing, storage and GI tract transition pose challenges indelivering probiotics for functional benefits
  8. 8. AgendaIdentifying the opportunityCore technologyComparison with selected established technologiesUnderstanding the intricaciesPath to commercialization8
  9. 9. ProbiolifeTM is a novel technology to improve the stability of probiotic bacteriaover a long period of storage at 25⁰CProbiotic strainGrowEncapsulateDryingMost of the common probiotic strains (e.g. Lactobacillus &Bifidobacterium) can be usedGrowth is under specially designed conditionEncapsulate in an optimised matrix – consisting of onlynatural ingredientsDrying using a special type of equipment with optimizedconditionsProbiolifeTM Technology - an insight
  10. 10. 45678910110 4 8 12 16 20 24 28 32 36 40 44 48LogCFU/GmDrySolidsStorage at 25ºC (Weeks)L. Casei CRL431 cells stored at ambient temperature maintained viability at over 1 billion livecells per gram after 9 months and over100 million live cells after 1 year.Storage StabilityImproved storage stability of probiotic bacteria at ambient temperature1 Billion
  11. 11. Understanding the ArchitectureBacteria L.casei CRL 431Encapsulation in matrixMatrix surface closer lookSEM images to elucidate the morphology of the matrix
  12. 12. ProbiolifeTM technology developed with L. casei CRL431, and then tested with a number ofLactobacillus and Bifidobacterium strains.For all the 8 strains tested ProbiolifeTM technology gave far better stability to the cells compared tofreeze drying during long term storage at ambient temperature.Lactobacillus acidophillus ATCC 4356 Lactobacillus rhamnosus ATCC 53103Bifidobacterium lactis BB12 Lactobacillus casei ATCC 393Validation of ProbiolifeTM using different probiotic strainsProbiolifeTM technology provided enhanced stability to the cells compared to freeze dryingduring long term storage at ambient temperature.Bifidobacterium lactis BB12 Lactobacillus casei ATCC 393
  13. 13. ProbiolifeTM technology provided enhanced protection to the cells during simulated gastro-intestinal transitSimulated gastric fluid at pH 2.0 with 0.32%pepsin.Free cells reduced to undetectable level after90 minEncapsulated cells reduced by 4.5 log cycleIncubation with 1% bile extract at pH 6.8.Free cells reduced by 2.7 log cycle.Encapsulated cells reduced by only 0.6 logcycleStability in gastric & intestinal fluid
  14. 14. The physical properties of the ProbiolifeTM ingredient is comparable with common dairyingredients making it feasible for wider range of applicationsProbiolifeTMWhole Milk PowderSkim Milk PowderProbiolifeTM physical propertiesD32 : 125.6 µmAw : 0.27Bulk Density: 470 Kg/m3ProbiolifeTMWhole Milk PowderSkim Milk PowderPhysical characterization
  15. 15. ProBioLife TM can be applied to a range of food productsProBioLifeTM powderShelf stable foods Shelf stable powderbeverageConform WHO/FAO recommendation 1 – 10 million live cells/gram....100 million cells per servingProBioLifeTM – application in consumer food productsRTD beveragesApplication usingdosing cap or like
  16. 16. 44.555.566.577.588.590 4 8 12 16 20 24LogCFU/GmStorage at 25ºC (Weeks)ProbiolifeTM fortified shelf stable food products maintained cell viability during storageShelf stability of ProbiolifeTM fortified productsL. casei CRL431 cells in encapsulated form were added to cereal bar, malted beverage and chocolate spread.The products were stored at 25ºC for 6 monthsThe loss in viable cell populations were recorded as 1.0, 0.2, 0.8 log CFU/gm for Powdereddrink, Chocolate spread and Cereal bars respectively.
  17. 17. AddedOriginalcell After After Final mixwater population 5 Min 30 Mintemperature(⁰C)temperature(LogCFU/gm)(LogCFU/gm)(LogCFU/gm) after 30 minAt 75⁰C 8.4 7.9 7.3 35At 65⁰C 8.4 8.3 7.9 25At 55⁰C 8.4 8.5 8.4 25Some consumers like to mix powder dairy beverage with hot water .....test confirmed highlevel of viable cells of L. casei after mixing with hot waterProBioLifeTM – application in powdered dairy beverage
  18. 18. Flavor Taste OverallAcceptabilityscoreTriangle test could not identify any difference between 2 chocolate flavour powderbeverage fortified with or without Probiolife ingredientNo negative impact of Probiolife ingredient fortification on the acceptability of powderbeverageProBioLife TM - application in powdered dairy beverageReconstituted chocolate flavoured powdered beverageReconstituted chocolate flavoured powdered beverage with ProBioLife ingredientProBioLifeTM ingredient does not impact the flavour of the food product
  19. 19. AgendaIdentifying the opportunityCore technologyComparison with selected established technologiesPath to commercialization19
  20. 20. Comparison of ProbiolifeTM technology Vs other established technology45678910110 4 8 12LogCFU/GmStorage at 25oC (weeks)Novel TechnologyCommercial freeze driedFreeze drying in samematrix as NTSpray drying in samematrix as NTCommercial freeze dried granules were most affected during ambient storageFreeze drying and spray drying process using ProbiolifeTM matrix showed slight improvementStabilization using ProbiolifeTM technology offered significantly better stabilityProbiolifeTM technology provided enhanced stability to bacterial cells compared to otherpopular stabilization technologies (freeze drying and spray drying )
  21. 21. AgendaIdentifying the opportunityCore technologyComparison with selected established technologiesPath to commercialization
  22. 22. ProBioLifeTM – cost comparable to many standard functional ingredientsProBioLifeTM powder competitive advantagesProbiotic strainGrowEncapsulateDryingAssumed: $500/kg asfreeze dried culture$10/kg ProBioLife powder(Assumption: $2.5/kg forprocessing and packaging)*Licence fee and capital investment not includedCompetitive advantage:Not strain specificNatural/GRASCost effective3 – 4 cents per serving for apowdered dairy based beverage
  23. 23. First provisional patent(Australia) – August 2010Second provisional patent(New Zealand) – April 2011PCT – August 2011Protecting the IP
  24. 24. ProBioLifeTM Consumer products companiesProBioLifeTM B2B ingredient companiesModel 1Model 2ProBioLifeTM Co-packerModel 3Consumer products companiesCulture supplyConsumer productscompanies/ingredient companiesModel 1:Abbott - Asia PacificGSK - IndiaPepsiCo – USA/ AMEAKraft – USASanitarium - AustraliaModel 2:Danisco - FinlandChr. Hansen – DenmarkDSM – Asia PacificBLIS – New Zealand(ABPL)Need to work out value chain for every modelCommercialization modelsThree potential business models
  25. 25.  Prof. Harjinder Singh-Co-Director, Riddet Institute , Massey University, Palmerston North, New Zealand Dr. Shantanu Das- Product Development Manager, Riddet Institute , Massey University, Palmerston North, New Zealand Mr. Arup Nag- Food Technologist, Riddet Institute , Massey University, Palmerston North, New Zealand Mr. Devastotra Poddar- PhD Student, Riddet Institute , Massey University, Palmerston North, New Zealand Mr. Sumon Saha- PGDip. Student, SEAT, Massey University, Palmerston North, New Zealand Mr. TK Vyas- PGDip. Student, IFNHH, Massey University, Palmerston North, New Zealand Mr. Jacob Lollike- Bachelor of Food Science ,University of Copenhagen ,Denmark Ms. Beibei Zhou- MSc ( Food Technology), Wageningen UR (University and research centre), Netherlands Ms. Esther Ang- Bachelor of Food Technology, Massey University, Singapore campus, Singapore Ms. Hui Kheng-Singapore Polytechnic, Singapore Ms. Marrie Camilie- Intern from France Prof. John Bronlund - Professor, SEAT, Massey University, Palmerston North, New Zealand Dr. Jon Palmer- Lecturer ,IFNHH, Massey University, Palmerston North, New ZealandContributors to the workAcknowledgementsProf. Paul Moughan for encouragement, motivation and valuable adviceMassey University Research Services for supportMr. Rick Gain and Mr. Richard Cartis for valuable adviceMr. Mark Ward for Encouragement and supportAJ PARK for help in PatentingManawatu Microscopy Services, IFNHH & SEAT for technical supportAll Riddet Institute staff members for help and support