Starter Lactic Acid Bacteria Research Articles

Effect of different starter cultures on the sensory properties and microbiological quality of Atole agrio, a fermented maize product

Maize is staple food in Central and Latin America, Southern and Eastern Africa, and fermentation processes are widely used to improve food safety and quality. Atole agrio is a typical Mexican fermented maize beverage, that, unlike many others, is made with a non-nixtamalized young maize. The aim of this research was to select endogenous lactic acid bacteria (LAB) starter cultures able to control atole agrio fermentation and preserve the sensory properties of spontaneously fermented atole agrio. Lactococcus lactis A1MS3 (12.6 ± 0.09 log cfu g−1) and Pediococcus pentosaceus S0l10 (11.7 ± 0.02 log cfu g−1), alone or as 1:1 v/v co-culture (12.5 ± 0.17 log cfu g−1) induced higher LAB counts than spontaneous fermentation (9.1 ± 0.11 log cfu−1) leading to fast acidification of the product. Starter cultures noticeably decreased Enterobacteriaceae, leading to microbiologically safer end product. Rapid sensory evaluation technique Flash profile (FP) differentiated the taste of products fermented with Lc. lactis alone or as co-culture with P. pentosaceus as sweet, tortilla and maize-like, compared to the acid and bitter taste of spontaneously fermented maize. We demonstrated the usability of FP in fermented maize products and showed, that atole agrio with enhanced sensory profile can be obtained by controlled fermentation.

Factors influencing the membrane fluidity and the impact on production of lactic acid bacteria starters.

Production of lactic acid bacteria starters for manufacturing food, probiotic, and chemical products requires the application of successive steps: fermentation, concentration, stabilization, and storage. Despite process optimization, losses of bacterial viability and functional activities are observed after stabilization and storage steps due to cell exposure to environmental stresses (thermal, osmotic, mechanical, and oxidative). Bacterial membrane is the primary target for injury and its damage is highly dependent on its physical properties and lipid organization. Membrane fluidity is a key property for maintaining cell functionality, and depends on lipid composition and cell environment. Extensive evidence has been reported on changes in membrane fatty acyl chains when modifying fermentation conditions. However, a deep characterization of membrane physical properties and their evolution following production processes is scarcely reported. Therefore, the aims of this mini-review are (i) to define the membrane fluidity and the methods used to assess it and (ii) to summarize the effect of environmental conditions on membrane fluidity and the resulting impact on the resistance of lactic acid bacteria to the stabilization processes. This will make it possible to highlight existing gaps of knowledge and opens up novel approaches for future investigations.

Growth prediction of two bacterial populations in co-culture with lactic acid bacteria.

The co-culture growth of Staphylococcus aureus, Escherichia coli and lactic acid bacteria starter culture in milk was quantitatively evaluated and modelled with a set of coupled differential equations originally proposed by Baranyi and Roberts and by Gimenez and Dalgaard (BR-GD model). The lactic acid bacteria starter culture showed the ability to induce an early stationary phase of both E. coli and S. aureus populations at different combination of temperature (ranging from 12 to 37 ℃) and lactic acid bacteria inocula (from approx. 103 to 106 CFU/ml). First, the prediction ability was performed only with parameters estimated from individual growth curves of E. coli, S. aureus and the lactic acid bacteria in milk (Dataset 1, 21 experiments). Subsequently, the model was extended with the average competition coefficients (E-BR-GD model) that represented quantitative relations among the populations. The prediction ability of this model was validated with the second dataset consisting of seven experiments. Results and also their statistical indices (accuracy and bias factors) showed that the E-BR-GD model improved growth prediction of all involved populations. Thus, the total root mean square error decreased from 0.457, 0.840 and 0.322 log CFU/ml (BR-GD model) to 0.290, 0.245 and 0.333 log CFU/ml (E-BR-GD) for S. aureus, E. coli and lactic acid bacteria, respectively. This approach in growth prediction of multiple competing microbial populations can be used in assessment of S. aureus and E. coli exposure from raw milk cheeses consumption and contribute to decision making in prevention of staphylococcal enterotoxin production.

PENGARUH STARTER BAKTERI ASAM LAKTAT PROBIOTIK TERHADAP PERUBAHAN KIMIAWI DAN MIKROBIOLOGIS RUSIP

Rusip is one of the fish fermentation products. Traditional rusip made by Bangka Belitung community do not have a specific standard which leads to the high variety ofquality. One of the efforts to improve the quality of rusip is the addition of lactic acid bacteria (LAB) as a starter culture. This research aims to study the effect of the addition of starter culture combinations on the chemical and microbiological changes of the rusip and toobtain the best combination of LAB starters in rusip fermentation. This research used anchovy fish (Stolephorus sp) and 5% ofLAB starter combination consisting of Lactobacillus plantarum SK(5) and L. plantarum NS(5), L. plantarum NS(5) and L. plantarum NS(9) and L. plantarum NS(9) and L. plantarum SK(5) 5%. In addition, 5% ofsalt (w/w) and 5% of palm sugar (w/w) were added. The fermenta-tions were conducted for 8 days. The research were conducted in three stages: 1) starter culture prepara-tion; 2) raw material (fish) preparation; and 3) application of starter culture during rusip fermentation. The results showed that raw materialscontained a total microbe of 3.0x104 CFU/g, non detectable levels ofLAB, 6.54of pH and 20.40 mg N/100g of total volatile bases (TVB). After fermentation, the rusip contained total LAB of7.64-9.24log CFU/g, 0.86-3.98% of total lactic acid,4.22-6.36 of pH, 0.32-1.39 of free amino acid and 5.34-5.44% of salt.The best rusip was produced bya combination of L. plantarum SK(5) and L. plantarum NS(5) and con-tained12.36 g/100g of total amino acid.

The effectiveness of lactic acid bacteria level on syneresis and acidity of fermented milk added carrot (Daucus carrota L) porridge

This experiment aimed to evaluate syneresis and acidity of fermented milk which is combined with varying levels of carrot porridge and lactic acid bacteria (LAB). There were 2 types of LAB used Lactobacillus acidophilus and Lactobacillus casei. The experiment carried out using a completely randomized design (CRD) of factorial patterns which is consisting of two factors. Factors were A: 2.5% and 5% level of starter LAB and B: three levels of carrot porridge (5, 10 and 15%). Variables measured were acidity (pH) and degree of syneresis. Data were analysed using analysis of variance (ANOVA) and the differences between treatment means were examined by Duncan Range Test (DMRT). The results showed that the pH very significantly (P <0.01) affected by LAB and carrot porridge level but syneresis significantly (P <0.05) effected only by level of carrot porridge. In conclusion that the best pH and syneresis resulting in 2.5 and 5% of starter at 5% carrot porridge level.

Impact of lactic acid bacteria starter cultures and hydrolytic enzymes on the characteristics of wholegrain crackers

This work studied the impact of bioprocessing with lactic acid bacteria starter cultures (dextran producing Weissella confusa and Lactobacillus plantarum) and proteolytic and xylanolytic enzymes on structural, chemical and sensory characteristics of wholegrain wheat crackers. Bioprocessing of flour increased free amino acids content (FAA) and water extractable arabinoxylans (WEAX). W. confusa fermentation increased FAA the most (0.46–0.48 g/100 g) compared to the control (0.22 g/100 g). Treatment with xylanase and fungal protease showed highest WEAX (2.29 g/100 g vs. 0.90 g/100 g). This cracker was also the thickest. Fermentation with W. confusa without dextran production decreased thickness, while dextran containing W. confusa samples were similar to the control. Cracker thickness and flour RVA-viscosities showed negative correlation. Non-dextran W. confusa increased final viscosity (1807 cP) and setback viscosity (1200 cP) compared to control flour (1318 cP and 885.5 cP). Both enzyme-types reduced viscosities: e.g. lowest final viscosity was 729.5 cP after protease treatment. Microscopy indicated enhanced gluten network especially after xylanase treatment. Enzyme-treated crackers had neutral flavor and soft structure, as well as increased tendency to break down in the mouth. Fermented crackers were hard and had pungent flavour. Acidification appeared to negate the possible positive impact of dextran on cracker quality.

Zoom on starter lactic acid bacteria development into oxytetracycline spiked ovine milk during the early acidification phase

Residues of antibiotics used in food-producing animals can be excreted in milk, affecting the technological performance of starter lactic acid bacteria (SLAB). SLAB development and acidification performance was studied in thermised ovine milk spiked with oxytetracycline (OTC) at maximum residue limit (MRL, 100 μg kg−1), during the early acidification phase. Late milk acidification and lower lactic acid concentration revealed an antibiotic effect mainly after 6 and 7 h from inoculation, with a 6 h delay to reach pH 5.6; the delay in SLAB development was observed by BactoScan™, while viable counts were not affected. Real-time PCR evidenced OTC effect for Streptococcus thermophilus and Lactobacillus helveticus, not for Lactobacillus delbrueckii subsp. lactis. A statistically significant impact of the antibiotic was observed, even at the maximum residue limit, with possible repercussions in technological processing and in human health, due to an increase of its concentration in cheese.

Potential application of lactic acid starters in the reduction of aflatoxin contamination in fermented sorghum-millet beverages

BackgroundAflatoxin contamination in traditionally fermented cereal-based beverages is a serious food safety challenge considering that commercialization of these products is rising. The challenge is aggravated by the fact that aflatoxin elimination from the food chain is almost impossible. This paper focuses on Obushera, a popular traditional spontaneously fermented sorghum – millet beverage from Uganda.MethodMold and total aflatoxin levels in cereal flours and Obushera from different markets in Kampala were determined. Ability of lactic acid bacteria (LAB) starters from Obushera; Lb. plantarum MNC 21, W. confusa MNC 20 and L. lactis MNC 24 to bind aflatoxin B1 (AFB1) was evaluated against Lb. rhamnosus yoba 2012 as the reference strain.ResultsMold counts in sorghum, millet and Obushera ranged between 0.0–2.4 log cfu/g, 2.0–6.5 log cfu/g and 2.0–5.5 log cfu/g, respectively. None of the flours complied with food safety standards for molds (maximum = 4 log cfu/g) while 88.0% of Obushera did (standard = maximum 1.3 log cfu/g). Total aflatoxin levels in sorghum, millet and Obushera were 22.3 ± 21.2 ppb, 9.9 ± 10.0 ppb and 10.4 ± 6.1 ppb, respectively. LAB bound 19.3–69.4% of AFB1 in a 1000 ppb solution with binding efficiency in the order of Lb. rhamnosus yoba 2012 = Lb. plantarum MNC 21 > W. confusa MNC 20 = L. lactis MNC 24. The LAB-AFB1complex remained stable under a series of washes with physiological saline.ConclusionThis study indicates that LAB with aflatoxin binding properties can be exploited for controlled cereal fermentations to improve food safety.

Application of lactic acid bacteria in green biorefineries.

Lactic acid bacteria (LAB) have extensive industrial applications as producers of lactic acid, as probiotics, as biocontrol agents and as biopreservatives. LAB play a large role in food fermentation and in silage processes, where crops such as grass, legumes, cereals or corn are fermented into high-moisture feed that is storable and can be used to feed cattle, sheep or other ruminants. LAB also have great applications within green biorefineries, with simultaneous production of protein-rich feed for monogastric animals, silage or feed pellets for ruminants and production of lactic acid or specific amino acids. In green biorefineries, fresh or ensiled wet biomass is mechanically fractionated into green juice and solid residues (press cake), where the plant juice, for example, can be used for production of lactic acid using LAB. In a process named 'ENLAC', recovery of protein and chlorophyll from silage by simultaneous lactic acid fermentation and enzyme hydrolysis has been developed. Furthermore, a process for protein recovery was recently developed by applying a specific LAB starter culture to green juice from freshly harvested crops. This paper focuses on reviewing LAB for their applications within biorefining of 'green' crops such as clover, alfalfa, grasses and other green plant materials.

Cheese yeasts.

Numerous traditionally aged cheeses are surface ripened and develop a biofilm, known as the cheese rind, on their surfaces. The rind of such cheeses comprises a complex community of bacterial and fungal species that are jointly responsible for the typical characteristics of the various cheese varieties. Surface ripening starts directly after brining with the rapid colonization of the cheese surface by yeasts. The initially dominant yeasts are acid and salt-tolerant and are capable of metabolizing the lactate produced by the starter lactic acid bacteria and of producing NH3 from amino acids. Both processes cause the pH of the cheese surface to rise dramatically. This so-called deacidification process enables the establishment of a salt-tolerant, Gram-positive bacterial community that is less acid-tolerant. Over the past decade, knowledge of yeast diversity in cheeses has increased considerably. The yeast species with the highest prevalence on surface-ripened cheeses are Debaryomyces hansenii and Geotrichum candidum, but up to 30 species can be found. In the cheese core, only lactose-fermenting yeasts, such as Kluyveromyces marxianus, are expected to grow. Yeasts are recognized as having an indispensable impact on the development of cheese flavour and texture because of their deacidifying, proteolytic, and/or lipolytic activity. Yeasts are used not only in the production of surface-ripened cheeses but also as adjunct cultures in the vat milk in order to modify ripening behaviour and flavour of the cheese. However, yeasts may also be responsible for spoilage of cheese, causing early blowing, off-flavour, brown discolouration, and other visible alterations of cheese.

Isolation of lactic acid bacteria from cattle rumen as starter in silage manufacture

The purpose of this research is to isolate lactic acid bacteria (LAB) found in rumen liquid of Bali cattle at slaughterhouse of Oeba City of Kupang, and to determine the benefits of lactic acid bacteria on the fermentation process of rice straw silage as animal feed based on the quality of rice straw given starter of lactic acid bacteria which is isolated from rumen fluid. Rumen fluid samples were cultured in De Man-Rogosa-Sharpe agar, isolated and identified by bacteria with Gram staining, catalase test and motility. Positive (LAB), bred with MRS (De Man-Rogosa-Sharpe) broth and added with palm sugar. The research treatment was differentiated according to the different dosage of LAB from each treatment (P0-P6). The results of the study can be said that LAB from rumen liquid can be used as a starter in rice straw silage with a damage level not reaching 50%.

Influence of the early bacterial biofilms developed on vats made with seven wood types on PDO Vastedda della valle del Belìce cheese characteristics

Early vat bacterial biofilms developed spontaneously through contact with whey have been characterized on seven wood types (Castanea sativa Miller, Cedrus libani, A. Rich., Prunus avium L., Fraxinus ornus L., Juglans regia L., Pinus nigra J.F. Arnold and Populus nigra L.). The present study aimed to evaluate the influence of these biofilms on the microbiological, chemical, physical and sensory characteristics of PDO Vastedda della valle del Belìce (VdB) cheese, processed traditionally from raw ewe's milk using wooden tools. To this purpose, the experimental cheeses after 15 d of refrigerated storage were examined. Lactic acid bacteria (LAB) populations dominated the microbial community of all samples. The species more frequently identified were Lactococcus lactis among starter LAB and Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus fermentum and Pediococcus pentosaceus among non starter LAB. Culture-independent analysis of microbiota diversity was performed by MiSeq Illumina that identified Streptococcus as major group followed by members of Enterobacteriaceae family, Lactococcus and Lactobacillus. Generally, the seven tree species did not negatively affect the physicochemical composition of VdB cheeses. Chestnut (both Sicilian and Calabrian) vats produced cheeses with significant lower hue angle (a*/b*) than other wood types. Among chemical parameters, significant variations were registered for aw, primary and secondary lipid oxidation state (significantly lower for the VdB cheeses produced with poplar wood), and volatile organic compounds (VOCs). The significant differences detected among the VOCs emitted from cheeses were not perceived by the panelists who recognized all cheeses from the different trials as similar. This study confirmed the suitability of cedar, cherry, ash, walnut, black pine and poplar as alternative woods to chestnut for the production of the wooden vats employed in cheese making for the Sicilian traditional dairy productions.

Randomized, double blind placebo-controlled pilot study of the antihypertensive effects of Grana Padano D.O.P. cheese consumption in mild - moderate hypertensive subjects.

Grana Padano, an Italian protected designation of origin (PDO) semi-fat cheese, undergoes a long ripening period during which the proteolysis carried out by natural starter lactic acid bacteria releases peptides having sustained angiotensin-converting enzyme (ACE)-inhibitory activity. The length (generally 3-8 amino acid residues) and the sequence of these peptides are responsible for their ability to elicit ACE-inhibitory activity. The aim of this study has been the evaluation of the effect of a daily dietary supplement consisting in a small amount (30 g/day) of Grana Padano cheese, in terms of the lowering of the blood pressure (BP) of mild-moderate hypertensive subjects. Thirty mild-moderate hypertensive patients, with BP values not on target (> 140 and/or > 90 mmHg) after at least 3 months of stable treatment were considered in this randomized, double-blind placebo-controlled cross-over study. All patients randomly received a dietary integration (30 g/day) of Grana Padano cheese or a placebo (made from flavored grated bread mixed with fats and salts in concentrations equal to those of the cheese). BP was evaluated at baseline and at the end of the active and placebo treatments (2 months each) by: - Office BP (OBP); - Automated Office BP (AOBP) using the BpTRU®, an automated oscillometric device that provides the average of multiple (n=6) blood pressure measurements; - Ambulatory Blood Pressure (ABP) 24 hour monitoring. Dietary integration with Grana Padano cheese resulted in a significant decrease in Office, Automated Office and Ambulatory BP. The mean decrease (vs. placebo) for 24-hour ABP was -3.5 mmHg for systolic and -2.4 mmHg for diastolic BP (p = 0.0063 and p = 0.0065, respectively). Daily dietary integration with 30 g of Grana Padano DOP cheese effectively reduces BP and may help mild-to-moderate hypertensive patients to reach a target BP.

Effect of addition of a starter cultures on physicochemical and sensory properties of “alheira”, a smoked sausage-like product

Introduction: “Alheira” is a traditional Portuguese smoked sausage-like product that can be manufactured with different types of meat and bread. Biopreservation of foods has gained special prominence due to the possibility of naturally controlling the growth of pathogenic and spoilage microorganisms, with a growing interest in the use of lactic acid bacteria (LAB) and their bacteriocins. Objectives: To ascertain the effect of adding fresh and dried starter LAB cultures (Lactobacillus plantarum ST153Ch) on the physicochemical and sensory characteristics of “Alheira”. Methods: All samples were produced at the industrial level. Sampling for analytical procedures was performed at 0, 15, 30, 45, 60, 75 and 90 days of storage at 4 C. Physicochemical characteristics (moisture content, pH, water activity (aw), instrumental hardness and adhesiveness, instrumental colour (L*, a*, b*)) and sensory evaluation, performed by 9 panellists regarding visual appearance, odour, texture and taste (11 attributes), were evaluated along the storage at 4 C. Results: Significant differences (p<0.05) were observed in all samples regarding moisture content, pH, aw, instrumental hardness, adhesiveness and in colour parameters, during the storage period. Conclusions: No significant differences in both inoculated “Alheira” samples compared to the control assay were found in sensory analysis until day 90, meaning that L. plantarum ST153Ch starter culture, fresh or dried, could be applied successfully until at least until day 75.

Ropy exopolysaccharide‐producing Bifidobacterium longum YS108R as a starter culture for fermented milk

SummaryBifidobacteria are generally inferior compared to traditional starter lactic acid bacteria (LAB) in terms of fermentation properties. The results of this study showed that the use of a ropy EPS producing strain Bifidobacterium longumYS108R during fermentation improved the physicochemical properties of fermented milk, including decreased whey syneresis and increased water‐holding capacity and apparent viscosity, however, another non‐ropy EPS producing strain B. longum C11A10B did not exert such effects. Furthermore, we found that synergistically fermented milk with YS108R and commercial starter could reduce the production of acetic acid, a metabolite of bifidobacteria, and scored better in terms of sensory evaluation, especially with regards to mouthfeel texture. Comparative genomic analysis revealed a single mutation in the protein‐tyrosine kinase gene of strain YS108R, which may relate to ropy phenotype. Our results indicate that ropy‐EPS producing B. longumYS108R is suitable as an adjunct culture for the manufacture of novel fermented dairy products.

PENINGKATAN BIOAVAIBILITAS PAKAN IKAN BERBASIS NABATI DENGAN AGEN FERMENTASI

Integration of simple biotechnology practice into conventional feed manufacturing methods is expected to improve the quality of artificial feed. Fermentation technology can be used as an alternative way to increase digestibility or bioavailability of artificial feed. The fermentation agent used is the starter of Lactic Acid Bacteria (LAB), Saccharomyces cerevisiae (Sc) and Rhizopus oligosporus (Ro). This experiment used Completely Randomized Design (CRD) with four treatments (F1; without fermentation agents, F2; BAL, F3; Sc and F4; Ro) and three replications so that the total experimental units were 12 experimental units. The variables measured included the physical and chemical quality of feed, the level of feed consumption and the growth parameters of tilapia. Fermented feed produced significant fish growth (P &lt;0.05) even with the same feed consumption level compared with the control. Breakdown of the nutrient content as a result of the fermentation process causes a far more efficient utilization.

Lactic Acid Bacteria Starter Participating in Hygienic Long Shelf-Life of the Plaa-Som Fermented Product

Consumptions of fermented vinegar made of fresh fruit juices have been increased dramatically due to their freshness, high vitamin content, and low caloric consumption. Unpasteurized fruit juice produced by pressing or squeezing of the fruits also have many diverse microflora which is normally present on the surface of fruits during harvest and postharvest processing and possibly include transport, storage, and processing. In the study, many microorganisms producing acid especially bacteria, fungi, and yeasts demonstrated the high acid production and using fruit juice as a substrate for their growth. Three acid producing bacteria were isolated and characterized for the acid production as well as applied for the fruit vinegar fermentation process.

Effect of High-Pressure Processing on Quality and Microbiological Properties of a Fermented Beverage Manufactured from Sweet Whey Throughout Refrigerated Storage

The production of fermented beverages is a promising way to valorize by-products of dairy manufacturing. However, the shelf-life of these products is often limited by the post-acidification process that occurs during storage. In this work, we manufactured a fermented beverage from sweet whey by using the starter lactic acid bacteria (SLAB) Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus. High-pressure processing (HPP) at 200 MPa for 10 min or 400 MPa for 1 min were applied after manufacturing. The aim of this study was to evaluate the effect of HPP on the quality of the beverage and on the behavior of the SLAB. Both high hydrostatic pressure treatments preserved flavor and texture attributes until 45 days post-HPP, without affecting chromatic parameters. Plate counts for both species were lower in HPP-treated beverages (HB) than in control beverages (CB), although treatment at 200 MPa maintained optimal amounts of total SLAB (6.6–7.9 log CFU/mL). Conversely, quantitative PCR (qPCR) and reverse transcription-qPCR (RT-qPCR) revealed that bacterial DNA or mRNA levels persisted after HPP (> 1.4 × 105 genome or cDNA copies/mL), even upon 400-MPa treatments. As a whole, this study indicated that HPP preserved the quality of the beverage until 45 days post-HPP, which is longer than the shelf-life of conventional fermented beverages obtained from milk. Moreover, our results obtained with these SLAB in a fermented dairy beverage upon HPP extend previous findings regarding the limitations of culture-dependent methods to assess microbial viability.

Generation of Bacteriophage-Insensitive Mutants of Streptococcus thermophilus via an Antisense RNA CRISPR-Cas Silencing Approach.

Predation of starter lactic acid bacteria such as Streptococcus thermophilus by bacteriophages is a persistent and costly problem in the dairy industry. CRISPR-mediated bacteriophage insensitive mutants (BIMs), while straightforward to generate and verify, can quickly be overcome by mutant phages. The aim of this study was to develop a tool allowing the generation of derivatives of commercial S. thermophilus strains which are resistant to phage attack through a non-CRISPR-mediated mechanism, with the objective of generating BIMs exhibiting stable resistance against a range of isolated lytic S. thermophilus phages. To achieve this, standard BIM generation was complemented by the use of the wild-type (WT) strain which had been transformed with an antisense mRNA-generating plasmid (targeting a crucial CRISPR-associated [cas] gene) in order to facilitate the generation of non-CRISPR-mediated BIMs. Phage sensitivity assays suggest that non-CRISPR-mediated BIMs exhibit some advantages compared to CRISPR-mediated BIMs derived from the same strain.IMPORTANCE The outlined approach reveals the presence of a powerful host-imposed barrier for phage infection in S. thermophilus Considering the detrimental economic consequences of phage infection in the dairy processing environment, the developed methodology has widespread applications, particularly where other methods may not be practical or effective in obtaining robust, phage-tolerant S. thermophilus starter strains.

Changes in folate characteristics and its identification in broccoli (Brassica oleracea Italica) extract fermented by Lactic Acid Bacteria Mixed Culture (LAB)

Broccoli (Brassica oleracea Italica) was fermented by cultures of lactic acid bacteria (LAB) as a potential source of natural folic acid. This study aimed to evalte characteristic changes and to identify folate compounds from broccoli extract, fermented by mixed LAB cultures (L. bulgaricus, S. thermophulus, L.acidophilus, Bd. bifidum). The formulation of broccoli extract was fermented with variation of LAB starter culture with concentrations of 10 and 20%(v/v), and the change of characteristic of folic acid compound during fermentation (0 to 48 hours) with an interval of 8 hours was evaluated. The results showed that the fermentation of broccoli extract with different concentration of LAB culture had an effect on the concentration of folic acid produced, as well as the change of concentration of folic acid during the fermentation time interval. The optimum condition was obtained based on the highest folic acid concentration of 6.74%, at culture concentration of 20% during 24 hour fermentation with the value of folic acid concentration of 72.11 μg/mL, pH value of 4.29, total sugars of 34.61%, total acids of 0, 97%, dissolved protein of 14.64 mg/mL and total LAB of log 13.02 + 0.05 cfu / ml.