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Storage Stability of Fermented Milk with Probiotic Monoculture and Transglutaminase

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Storage Stability of Fermented Milk with Probiotic Monoculture and Transglutaminase Food Microbiology and Safety Czech Journal of Food Sciences, 37, 2019 (5): 332–337 https://doi.org/10.17221/22/2019-CJFS Storage stability of fermented milk with probiotic monoculture and transglutaminase Izabela Dmytrów1*, Anna Mituniewicz-Małek1, Małgorzata Ziarno2, Jerzy Balejko3 1Department of Dairy Technology and Food Storage, Faculty of Food Sciences and Fisheries, West Pomeranian University of Technology Szczecin, Poland 2Department of Biotechnology, Microbiology and Food Evaluation, Faculty of Food Sciences, Division of Milk Biotechnology, Warsaw University of Life Sciences, Poland 3Department of Food Process Engineering, Faculty of Food Sciences and Fisheries, West Pomeranian University of Technology Szczecin, Poland *Corresponding author: [email protected] Citation: Dmytrów I., Mituniewicz-Małek A., Ziarno M., Balejko J. (2019): Storage stability of fermented milk with probiotic monoculture and transglutaminase. Czech J. Food Sci., 37: 332–337. Abstract: The effect of microbial transglutaminase on selected physicochemical and organoleptic characteristics and viability of probiotic bacteria in fermented milk inoculated with probiotic monoculture (Lactobacillus acido- philus LA 5 or Bifidobacterium bifidum BB 12) was analysed. Four types of samples were prepared: (1) fermented milk inoculated with Lactobacillus acidophilus LA 5, (2) fermented milk inoculated with Bifidobacterium bifidum BB 12, (3) fermented milk produced from milk previously treated with mTGase and inoculated with Lactobacillus acidophilus LA 5, (4) and fermented milk produced from milk previously treated with mTGase and inoculated with Bifidobacterium bifidum strain BB 12. The samples were analysed after the st1 , 7th and 14th day of storage at 5 ± 1°C. It has been found that the use of microbial transglutaminase for the production of fermented milk inoculated with monoculture affected its viscosity, hardness, acetaldehyde content and increased the viability of probiotic bacteria. The enzyme activity resulted in an significant decrease in the titratable acidity of the experimental products, positi- vely affected viscosity, the viability of probiotic bacteria and the organoleptic properties of fermented milk. Keywords: acidity; enzyme; fermentation; hardness; viscosity Fermented milk is considered by nutritionists soft and short-lasting, especially in case of a single as having high nutritional value and positive bioactive probiotic strain (Zaręba et al. 2008). effects, usually reinforced by the addition of prebi- A recently used method for shaping the tex- otic ingredients and probiotic bacteria. The genera ture of fermented milk is based on the applica- Lactobacillus and Bifidobacterium are used the most tion of transglutaminase (protein-glutamine amine often as probiotic bacteria in this type of products γ-glutamyltransferase). This enzyme catalyses acyl (Nowak et al. 2010) but the fermented milk should transfer reactions between γ-amide groups of glu- contain at least 106 CFU of labelled (i.e., probiotic) tamine in proteins, as donors of the acyl groups, and bacteria per millilitre (Wang et al. 2012). Probiotic primary amines, which are acceptors of these groups. bacterial cells do not have the ability to typically fer- Enzyme actions cause changes in the structure of ment milk. A clot produced by them is usually very proteins and peptides, of which the most important Supported by the West Pomeranian University of Technology in Szczecin, Grant No. 518-08-57-3181-01/18. 332 Czech Journal of Food Sciences, 37, 2019 (5): 332–337 Food Microbiology and Safety https://doi.org/10.17221/22/2019-CJFS is the formation of crosslinks that alter the ability microorganism Streptoverticillium mobaraense (1%) to retain water, solubility, viscosity, elasticity, smell and 99% of maltodextrin. It was used in the original or colour of the product. Microbial transglutaminase form, without any further purification (Dmytrów (mTGase) is a single polypeptide chain with a molecu- et al. 2010). lar weight of 38,000 Da, consisting of 331 amino acids, Fermented milk – processing conditions. The with the isoelectric point at a pH of 8 to 9. Although pasteurized milk was heated to 40°C and divided improvement in the rheological characteristics of into 4 batches (each of 4000 ml). Two of them probiotic fermented milk can be achieved by using (2 × 4000 ml) were enriched with 0.02% of Activa mTGase, the possible effects of this treatment on the MP® and then were incubated for 2 h at 40°C. After viability of probiotic strains should be kept in mind. the incubation, the processed milk was heated at 80°C In the subject literature, there is no comprehensive for 1 min to inactivate the enzyme. After cooling information on the effect of cross-linking enzyme to 40°C, each batch of milk treated with mTGase on the quality of fermented milk inoculated with was inoculated with 2.5% of activated starter con- probiotic monoculture and on the viability of these taining single probiotic strains, i.e., L. acidophilus strains in the end-product. Therefore, in this study LA 5 and B. animalis ssp. lactis BB 12, respectively. we assessed the impact of mTGase on the organo- The other two batches of milk were inoculated only leptic characteristics, acidity, hardness, viscosity and with the listed potentially probiotic strains (variants acetaldehyde content in fermented milk contain- without the addition of mTGase). Incubation of all ing probiotic monoculture such as Lactobacillus variants of fermented milk was carried out at 42°C. acidophilus LA 5 and Bifidobacterium animalis ssp. The end of the fermentation was indicated by the pH lactis BB 12. and fermentation curve set in the culture specifica- tion. The samples were cooled down to 5 ± 1°C and stored at the same temperature for 14 days. Four MATERIAL AND METHODS types of samples of fermented milk were obtained: inoculated with L. acidophilus LA 5 (LA), inoculated Materials. The raw material was homogenised with B. animalis ssp. lactis BB 12 (BB), treated with (15 MPa, 55°C) and pasteurized at a high temperature mTGase and inoculated with L. acidophilus LA 5 (LA- (85°C, 30 s) cow’s milk, containing 3.2% fat, 3.0% TG), and treated with mTGase and inoculated with protein and 4.8% lactose. B. animalis ssp. lactis BB 12 (BB-TG). The samples Probiotic cultures. The experimental fermented were tested after 1st, 7th and 14th days of storage milk was produced not with the use of conventional at 5 ± 1°C. starter cultures commonly applied in the production Physicochemical analysis. The following charac- of various kinds of milk-fermented beverages but teristics were analysed in the samples: acetaldehyde only with probiotic freeze-dried DVS type monocul- content using diffusion method with hydrochloride tures of Lactobacillus acidophilus LA 5 and Bifido- hydrazine in Conway Chambers (Lees & Jago 1969), bacterium animalis ssp. lactis BB 12 manufactured titratable acidity in Soxhlet Henkel degrees (AOAC, by Chr. Hansen (Hoersholm, Denmark). The strains 2000) and active acidity were measured with a pH B. animalis ssp. lactis BB 12 and L. acidophilus LA 5 meter (IQ 150; Spectrum Technologies, UK). The are among Chr. Hansen’s best clinically documented analysis was performed in 6 replicates. probiotic strains. They have been tested in numerous Hardness measurement. Texture profile analysis clinical studies and have demonstrated health benefits was performed using a TA.XT Plus texture analyser in relation to gastrointestinal health. Reviving and with a computer set (Stable Micro System Ltd., UK). activating starters were performed as per the instruc- The samples were penetrated with a pressure of 1G tions of Technological Production Starter, developed and a velocity of 5 m/s up to a depth of 25 mm. The by Bielecka (1984) in non-fat milk (0% fat). diameter of the aluminium pin was 20 mm. The analysis ® Transglutaminase. Activa MP (E.C. 2.3.2.13) was performed at 10°C in 6 replicates. (Ajinomoto Co. Inc., Japan) was used as an additive Viscosity measurement. Viscosity measurement during the production of two variants of fermented of stirred fermented milk was carried out at 20 milk. This enzyme preparation, recommended for ± 1°C using the measurement system composed of cross-linking of milk proteins, is composed of Ca+2 double gap concentric cylinders in TA Instruments independent transglutaminase derived from the AR 2000 rheometer. The apparent viscosity of the 333 334 ple BB. case sam the of most significantthe – in the found variants was the milk all of pH fermented of the in period, storage first last and the of a decrease days experimental products.the the Whencomparing an significant decrease acidity of titratable inthe activity resulted in mTGase BB. in smallest the and LA, sample the in largest the – 1) (Table samples of increasecant variants acidity ofallthe titratable inthe tests was significancethe statistical all of test. The donebymeans post-hoc Dunn’s were ofthe test. pairwise The comparisons Kruskal-Wallis of the Organoleptic assessment was analysed bymeans t-test). means independent and (Student’s pendent de 2 in differences the testsdetermine and ures to withrepeated meascarried 2-way by ANOVA out performed assessmentleptic were series. in three mouths. Allphysicochemicalanalysis andorgano test stand anddistilled rinse to water their a separate was free odours; foreign ofany each panellist had evaluation The was score. carried room a in that out was 5 and best was 1 scale score the worst where the evaluated were consistency samples of 5-point a on and taste, appearance, smell ISO to The ing (1998). evaluation milk,organoleptic fermented of accord conductedwere byparticipants ( product units(CFU/g). per the of g 1 number ofcolony-forming the convertedto were KGaA,Anaerocult A(Merck Germany).results The equipped jar husing 37°C/72 anaerobic an at with was used cultures. incubated the were plates for The KGaA, purchased(Merck Germany) from Merck MRS (EN ISOagar The mediumsamples 6887-5:2010). threeanalysed replicatesofeachreplicates for ofthe independent method parallel withtwo sical plate clas the by milk microflora fermented biotic the in pro- number ofthe cell included ofthe determination the performed were ments thrice.
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