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BARENTZ INGREDIENTS Barentz Ingredients is the Technology Center for development of innovative and customized solutions for Food Industry, within the Barentz Group. Competency Texturizing of food systems Bindingsystems Injection/tumble solutions Proteins / Hydrolysates Taste solutions Natural preservatives Application Development and Coaching

Products and Technology Misocarine LR Product and Technology Properties and Application Labeling and Food Law

Product and Technology Misocarine LR Misocarine LR is a naturally fermented liquid product, which inhibits growth of bacteria and moulds and optimizes the taste profile of meat products Spices and glucose syrup are naturally fermented by lactic acid bacteria and acetic acid bacteria Misocarine LR is labeled as spice extract or onion extract

Production Process spices (onion) / glucose syrup supplements enzymatic hydrolysis fermentation broth culture 1 fermentation fermented spices (lactic acid) culture 2/3 fermentation fermented spices (acetic acid) neutralisation (sodium hydroxide) heat inactivation separation of biomass fermentation supernatant clarification concentration MISOCARINE LR

Properties of Misocarine LR Growth Inhibition During fermentation, lactic acid bacteria and acetic acid bacteria produce small molecular organic acids and antimicrobial substances. Those substances accumulate in Misocarine LR. Modes of bacterial growth inhibition Organic acids, antimicrobial substances and their salts can penetrate bacterial cells and molds inactivate metabolic acitivity of bacteria and moulds even at neutral ph values (feedback inhibition / accumulation of salts) reduce the water activity (a w -value)

Reduction of water activity* Properties of Misocarine LR Reduction of water activity 0,14 0,12 0,1 0,08 0,06 0,04 0,02 0 Salt sodium lactate (60%) Misocarine LR * dosage: 20 g/kg Misocarine LR is able to reduce the water activity of meat products

Reduction of water activity* Properties of Misocarine LR Reduction of water activity 0,14 0,12 0,1 0,08 0,06 0,04 0,02 0 Salt sodium lactate (60%) Misocarine LR * dosage: 20 g/kg Misocarine LR is able to reduce the water activity of meat products

Properties of Misocarine LR Mode of Inhibition at neutral ph ph 6.0 Organic acids are fully undissociated at neutral ph values ph 7.0 Salts of organic acids are able to inhibit the metabolism of bacteria and moulds

Properties of Misocarine LR Mode of Inhibition at acidic ph + + ph 4.5 Organic acids are fully dissociated at ph values below 4.5 + + + + + ph 7.0 + + + ph 4.5 Organic acids lower the internal ph value of bacteria and moulds at low external ph

Application of Misocarine LR Tested, approved Applications / Meat products Product Fresh meat (minced meat) Boiled sausage (Frankfurter) Cooked ham (sliced product) Cured ham (ham dice) Fermented sausage (salami) Reason for application Shelf life extention (2-3 days) Shelf life min. 30 days Shelf life min. 30 days Shelf life extention, product protection Shelf life extention, product protection Dosage 2 3%

Keime / g Application of Misocarine LR Fresh minced meat is mixed with 2 % additive (Na-Lactate or Misocarine LR) Samples are incubated at 8 C for 11 days The total bacterial count is measured each second day 1,0E+06 1,0E+05 Total count Gesamtkeimzahl mesophilic bacteria Wasser Water (Kontrolle) (control) FG Misocarine L100 2% LR 2 % 60 60 % % Na- NaL Lactat 2 % 2% 1,0E+04 1,0E+03 1 3 5 7 9 11 Tage Growth inhibition of bacteria can be extended for 6 days by 2 % Misocarine LR

Keime / g Application of Misocarine LR Fresh minced meat is mixed with 2 % additive (Na-Lactate or Misocarine LR) Samples are incubated at 8 C for 11 days The number of lactic acid bacteria is measured each second day 1,0E+06 1,0E+05 Lactic acid Milchsäure-Bildner bacteria Wasser Water (control) Misocarine LR 2 % FG L100 2% 60 % NaL 2 % 1,0E+04 1,0E+03 1 3 5 7 11 Tage Growth Inhibition of lactic acid bacteria can be extended for 5 days by 2 % Misocarine LR

Keime / g Application of Misocarine LR Fresh minced meat is mixed with 2 % additive (Na-Lactate or Misocarine LR) Samples are incubated at 8 C for 11 days The number of yeast and moulds is measured each second day Gesamtkeimzahl Yeast Hefen and und moulds Schimmel 1,0E+06 1,0E+05 Wasser Water (control) (Kontrolle) Misocarine LR 2 % FG L100 2% 60 % NaL 2 % 1,0E+04 1,0E+03 1 3 5 7 11 Tage Growth Inhibition of yeast and moulds can be Extended for 2 days by 2 % Misocarine LR

Application of Misocarine LR Test result Product: Frankfurter Sausages, 20 g/kg Storage Temperature: 8 C Test period: 30 days Parameter method T0 T30 critical limit DGHM* mesophilic aerobic bacteria ASU L 06.00-1.8, PCA/30 C/48 h <10.000 30.000 50.000 E. coli ASU L 01.00-3, Rapid E.coli/37 C/24 h <10 <10 10 Enterobacteriaceae ASU L 06.00-24, VRBD/30 C/48 h, anaerob <100 <100 100 coagulase positive Staphylococcus ASU L 00.00-55, BP/37 C/48 h <10 <10 10 Lactic acid bacteria ASU L 06.00-35, 30 C /72 h <10.000 <10.000 50000 yeasts ASU L 01.00-37, YGC/25 C/72-120 h <1.000 <1.000 10000 Listeria monocytogenes ASU L 00.00-22 <100 <100 100 Salmonella ASU L 00.00-98 n.n./25 g n.n./25 g n.n./25 g spores of sulfit reducing clostridia ASU L 06.00-39, TSC/37 C/48 h anaerob <100 <100 100 *Deutsche Gesellschaft für Hygiene und Mikrobiologie critical value for Frankfurter sausages arotop food & environment GmbH

Application of Misocarine LR Test result Product: Sliced Cooked ham, 20 g/kg Storage Temperature: 8 C Test period: 30 days Parameter method T0 T30 critical limit DGHM* mesophilic aerobic bacteria ASU L 06.00-1.8, PCA/30 C/48 h <10.000 20.000 5.000.000 E. coli ASU L 01.00-3, Rapid E.coli/37 C/24 h <10 <10 10 Enterobacteriaceae ASU L 06.00-24, VRBD/30 C/48 h, anaerob <100 300 1.000 coagulase positive Staphylococcus ASU L 00.00-55, BP/37 C/48 h <10 10 10 Lactic acid bacteria ASU L 06.00-35, 30 C /72 h 20.000 4.000.000 5.000.000 yeasts ASU L 01.00-37, YGC/25 C/72-120 h <1.000 <1.000 10.000 Listeria monocytogenes ASU L 00.00-22 <100 <100 100 Salmonella ASU L 00.00-98 n.n./25 g n.n./25 g n.n./25 g spores of sulfit reducing clostridia ASU L 06.00-39, TSC/37 C/48 h anaerob <100 <100 100 *Deutsche Gesellschaft für Hygiene und Mikrobiologie critical value for Frankfurter sausages arotop food & environment GmbH

Application of Misocarine LR Possible Labeling/FoodLaw EG Aroma Regulation Nr. 1334/2008 and Reg. 2000/113/EG; 6 Abs. 5 LMKV Professional opinion: Spice Extract Onion Extract Aroma Extract Aroma Natural Aroma Natural Onion Extract with other Natural Extracts

Misocarine LR Declaration: Spice extraxt E-number free Extent the shelf life with 2-3 days compared to NaL 60% Lower dosage than NaL 60%: only 2% Patented formula Authosrised within the EU food legislation

Barentz Ingredients Technology and Application Partner for Technology and Application development Support in Production plants Support in Labeling and Food Law Studies Product and Technology development