## 4 Discussion  
The isolation process of lactobacilli was carried out in a dilution process up to 10-8, a multi-step dilution method aimed at reducing the colony density on the medium to facilitate the isolation process [26]. Isolates were grown using MRSA medium supplemented with 1% CaCO\({}_{3}\) and incubated at 37 \({}^{\circ}\)C for 48 h. The deMann, Rogosa, and Sharpe agar (MRSA) was a selective medium for growing lactic acid bacteria. When 1% CaCO\({}_{3}\) was added to the medium, lactobacillus colonies grew, as indicated by the presence of clear zones around the colonies [27]. The isolation resulted in five isolates: DSK 1, DSK 2, DSK 3, DSK 4, and DSK 5.  
The five isolates were tested for antimicrobial activity and microscopic morphology to generate BAL candidates for bacteriocin production. The results indicated DSK 1 as the selected isolate with the highest antibacterial activity. Excessive acidic conditions cause lipopolysaccharide-aschardie dissolution, pore formation that impairs energy synthesis and cell wall permeability, and ultimately leads to pathogen death. DSK 1 is a selected isolate to be further molecularly identified. A molecularly specific test was targeted to confirm dadith's specific strain of bacteria. Dadith products include LABs of various strains. Bacterial strains common in dadith include _Bifidobacterium_, _Lactobacillus_, _Lactococcus_, _Streptococcus_, and _Enterococcus_[28]. Molecularly, Lactobacillus was considered to determine specific strains. As a result of analysis, the lactic acid bacterium was found to be the _Lactobacillus pentosus_ 124-2 strain.  
Based on the results, the pure bacteriocin produced by gel filtration chromatography exhibited higher specific activity than that exhibited in the previous two methods. An increase in specific activity indicates an increase in bacteriocin purity and a constant value indicates that the bacteriocin is completely pure [29]. However, the yield values were too low and should be increased by optimizing production conditions, such as optimizing medium components and non-culturing conditions. Large-scale production of bacterios requires a purification process that achieves >50% yield at 90-fold purification speed [30]. Some bacterios have similar properties, allowing a comparison of different purification processes.  
Antibacterial activity results showed that the bacteriocin _L. pentosus_ was more resistant to the Gram-negative bacterium _Salmonella_ sp. than to the Gram-positive _S. aureus_. Based on comparison, most bacterios produced by Lactobacillus species are only good against Gram-positive bacteria. Bacteriocin-derived _L. pentosus_ was comparable to Nissin, which can only inhibit Gram-positive bacteria, and to _L. plantarum's bacteriocin, which can fight both Gram-positive and harmful bacteria, but it was unable to inhibit fungal growth. In contrast, some _L. pentosus_ bacterios have been reported to be able to inhibit fungi such as Candida albicans, thus providing _L. pentosus_ bacterios with the potential for broader applications [6]. However, bacterios have less antibacterial activity than antibiotics, but due to increasing antibiotic resistance, bacterios have yet to be developed and further studied.  
Various factors affect the activity of bacterios, including temperature, pH, salinity, and surfactants. Characterization of changes in temperature, pH, concentration of NaCl, and surfactant type determined the activity potential of a bacteriocin as an antibacterial agent, its ability to meet the required antibiotic susceptibility pattern, and its inhibitory activity against pathogenic microorganisms. An evaluation test was developed [31]. The production of bacterios used as natural preservatives requires knowledge of their properties and viability under certain conditions in order to understand basic information about the function and specific properties of bacterios in their applications.