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Evaluation of Locally-Isolated Lactic Acid Bacteria as Mozzarella Starter Cultures

Abigail A. Mercado and Maris Hazel A. Cura

University of the Philippines Rural High School
Lactic acid bacteria (LAB) contribute to the texture, flavor, and quality of cheese. It also assists in lowering the pH, coagulation, syneresis, prevention of spoilage and growth of pathogenic bacteria during cheese production. The general objective of the study is to screen and evaluate the 17 locally-isolated LAB, obtained from Central Luzon by Gervasio and Lim (2007), and three reference strains Lactobacillus paracasei 1Y9 from commercial Yakult, Lactobacillus casei 1065, and Lactobacillus delbrueckii 1010 as starter cultures for Mozzarella.
Table 2. Simplified costing (in pesos) of Mozzarella preparation The coagulated milk was cut into 1cm cubes after addition of starter culture, diluted glacial acetic acid, and microbial rennet.



3F10:1Y9 100.00 8.00 0.36

2Li carabaos 100.00 milk 30ml lactic 8.00 starter culture 100ml diluted acetic acid (1.8ml/100ml water) 1Li 2% brine (rock salt) 20ml microbial rennet Total cost Recovered cheese, g Cost per gram cheese Cost per 200g 0.36

S.thermophilu s 100.00 25.00 0.36

Seventeen locally-isolated LAB were tested for diacetyl and percent lactic acid production or titratable acidity. The chosen isolate combinations such as 4E3:1Y9 and 3F10:1Y9 were used as starter cultures for Mozzarella production. An imported commercial starter, Streptococcus thermophilus was also used in Mozzarella production. Sensory evaluation of the prepared Mozzarella cheeses were conducted and compared with ARLA, an imported Mozzarella cheese produced from Denmark. Results showed that Mozzarella cheeses produced using 4E3:1Y9 and 3F10:1Y9 were relatively cheaper than the Mozzarella cheese produced using S. thermophilus. Furthermore, these Mozzarella cheeses were even much cheaper than the imported ARLA Mozzarella. Sensory evaluation of the Mozzarella cheeses revealed that there was no significant difference between the prepared cheeses and the commercial cheese ARLA in term of flavor, texture, and general acceptability

Toasted Mozzarella and bread were coded with 3digit numbers and were evaluated by 10 taste panelists. (A = ARLA commercial Mozzarella, B = 3F10:1Y9, C = 4E3:1Y9, D = Streptococcus thermophilus)

0.14 9.60

0.14 9.60

0.14 9.60

118.10 445 0.27 54

118.10 436 0.27 54

135.10 368 0.37 74

*Microbial rennet = 480 pesos per Li; rock salt = 7 pesos per kg, ARLA Mozzarella = 165 pesos per 200g

Results and Discussion

Using microscopic observation, reference strains 1Y9, 1065, and 1010 proved to be Gram positive rods, while the 17 LAB isolates were Gram positive cocci or spherical cells. The only isolate that produced diacetyl was 1Y9, and this was paired with 3F10 and 4E3, which had the highest titratable acidity of 0.46 and 0.45%, respectively. No significant differences were observed between the prepared Mozzarella cheeses and ARLA commercial cheese in terms of flavor, texture, and general acceptability. Significant differences were observed between the prepared Mozzarella cheeses and ARLA commercial cheese in terms of color, aroma, and saltiness at probability levels of less than 5%
Table 1. Diacetyl production and percent titratable acidity of 17 locally-isolated lactic acid bacteria and three reference strains. Table 3. Summary of the means _/ with respect to sensory qualities of Mozzarella cheeses produced using local starters (4E3:1Y9 and 3F10:1Y9), imported S. thermophilus, and ARLA commercial cheese Cheese Samples Color Criteria Aroma Saltiness Flavor Texture General Acceptability 3.700A 4.800A 3.800A 4.600A 3.500A 5.100A 6.100A 6.300A 6.000A

4E3 : 3.400B 5.200B 3.200B 1Y9 3F10 : 3.500B 5.300B 3.800A 1Y9 S. 3.500B 5.100B 4.000A Thermophilus ARLA

Glycerol stocks of LAB isolated and reference strains were revived and their purity were ascertained by Gram staining and observation under the microscope. Active cultures of LAB were tested for diacetyl and lactic acid production (Davide, 1980). Selected starter cultures and commercial S. thermophilus were used for Mozzarella production. Cost analysis of Mozzarella production from 2 Li carabaos milk and sensory evaluation of the Mozzarella cheeses were also done.

5.200A 5.200A 3.800A

4.100A 4.800A


Isolate Code Diacetyl Production 2A8 3B2 4E1 4E2 4E3 4E4 4E5 4E6 4E7 4E9 4E10 3F8 3F10 3G1 3G8 3G9 3G10 *Reference Strains: 1Y9 1065 1010 ___ ___ ___ ___ ___ ___ ___ ___ ___ ___ ___ ___ ___ ___ ___ ___ ___

The lactic acid bacteria inoculum was added to the carabaos milk

Titratable Acidity (% lactic acid) 0.26 0.28 0.27 0.27 0.46 0.26 0.26 0.29 0.26 0.33 0.26 0.27 0.45 0.43 0.28 0.26 0.26

of Variance (ANOVA). Duncans Multiple range Test (DMRT). Means with common letters are not significantly different.

Conclusion and Recommendations

Mozzarella cheeses produced using starter cultures 3F10:1Y9 (0.27 cents/g) and 4E3:1Y9 (0.27 cents/g) were relatively cheaper than the imported S. thermophilus starter culture (0.37 cents/g). These prepared cheeses were also cheaper than the imported Mozzarella from Denmark (ARLA) which cost 0.82 cents/g or 165 pesos/200g. No significant differences were observed between the prepared cheeses and ARLA commercial cheese in terms of flavor, texture, and general acceptability. It is recommended that complete biochemical and physiological tests of local isolates 3F10 and 4E3 be done to know their identity. Research to better define the relationship between lactic acid physiology and flavor development is suggested.

BIOTECH microbial rennet produced from local isolate Rhizopus chinensis was also added

+++ ___ ___

0.36 0.21 0.25

Davide, C.V. 1980. Handbook and Training Manual. Small Scale Production Technology for Starter Culture, Yoghurt, Cottage Cheese, and Quality Testing Gervasio A.T.R. and V.M.T. Lim. 2007. Undergradute thesis. Probiotic Characterization of Bacteriocinogenic LAB Isolated from Fermented Foods and Selected Areas of Central Luzon. U.P. Manila. 105pp.

* L. paracasei 1Y9, L. casei 1065 ,L. delbrueckii 1010