scholarly journals IDENTIFICATION OF GABA-PRODUCING LACTIC ACID BACTERIA FROM CINCALOK FERMENTATION BASED ON THE 16S RRNA AND GAD GENES

2021 ◽  
pp. 102-106
Author(s):  
ADHITYA NAUFAL PRIBADHI ◽  
ENDANG KUSDIYANTINI ◽  
REJEKI SITI FERNIAH
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Target Gene ◽  
Rrna Gene ◽  
Identification Analysis ◽  
16 S Rrna ◽  
The 16S Rrna Gene ◽  
Weissella Confusa

Objective: The research to identify LAB using 16S rRNA potential as high produce GABA and design primer can amplify that gad gene. Methods: Isolation genomic form LAB, molecular identification based 16S rRNA, design primer use primer3plus, and use application serial cloner to ensure the primer can amplify to target gene. Results: That have been carried out based on the analysis of the 16 S rRNA gene have the highest similarity to Weissella confusa strain JCM 1093 with a similarity of 98.38%, while the results of the analysis of the gad gene with several primers that have been designed are not able to amplify the gad gene owned by W. confusa. Conclusion: The results of the analysis based on the 16S rRNA gene for lactic acid bacteria were obtained by Weissella confusa. However, for the results of identification analysis based on the gad gene, the designed primers were unable to amplify the gad gene in W. confusa.

scholarly journals Diversity analysis, identification, and bioprospecting of Lactic Acid Bacteria (LAB) isolated from Sumbawa horse milk

2021 ◽  
Vol 22 (6) ◽  
Author(s):  
KHADIJAH ALLIYA FIDIEN ◽  
Baso Manguntungi ◽  
LINDA SUKMARINI ◽  
APON ZAENAL MUSTOPA ◽  
LITA TRIRATNA ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Lactococcus Lactis ◽  
Rrna Gene ◽  
Diversity Analysis ◽  
Indigenous Bacteria ◽  
Dpph Method ◽  
The 16S Rrna Gene

Abstract. Fidien KA, Manguntungi B, Sukmarini L, Mustopa AZ, Triratna L, Fatimah, Kusdianawati. 2021. Diversity analysis, identification, and bioprospecting of Lactic Acid Bacteria (LAB) isolated from Sumbawa horse milk. Biodiversitas 22: 3333-3340. Sumbawa horse milk has a probiotic potential because of the presence of Lactic Acid Bacteria (LAB). The LAB present in Sumbawa horse milk has been reported to have antimicrobial activities against pathogenic bacteria, including Staphylococcus epidermidis, Staphylococcus aureus, Escherichia coli, and Vibrio cholerae. However, the potential of LAB from Sumbawa horse milk as antioxidant and antidiabetic is still unexplored. Studies related to the diversity of indigenous bacteria in Sumbawa horse milk based on metagenomic analysis have not been widely studied either. Therefore, this study aimed to determine the diversity of species of indigenous bacteria in Sumbawa horse milk and to identify LAB bioprospecting from Sumbawa horse milk. The diversity of indigenous bacterial species was investigated by the 16S rRNA gene-targeted metagenomic approach from bacterial DNA isolated from Sumbawa horse milk. The identification of LAB was also carried out by the 16S rRNA gene identification method. LAB bioprospecting on antioxidant activity was determined using the DPPH method, while the antidiabetic activity was measured using the ?-glucosidase inhibition assay. The diversity analysis of indigenous bacteria based on 16S rRNA gene-based metagenomic revealed at least 7 phyla were relatively abundant in Sumbawa horse milk. The greatest abundance was shown by the phylum Proteobacteria (0.641%) and Firmicutes (0.327%). Enterococcus durans (39.01%) was the species that had the highest abundance in Sumbawa horse milk, followed by Lactococcus garvieae (30.13%) and Lactococcus lactis (19.85%). Moreover, based on the identification of the 16S rRNA gene, eight LAB isolates had similarities to bacterial strains, including Enterococcus faecium DSM 20477, E. faecium NBRC 100486, E. faecium ATCC 19434, E. durans 98D, E. faecalis ATCC 19433, E. faecalis NRBC 100480, Lactococcus lactis subsp. hordniae NBRC 100931 and L. garvieae JCM 10343 with similarity levels of more than 98%. In terms of LAB bioprospecting, the antioxidant assay showed the highest DPPH radical binding activity by L. garvieae L.22PR (43%). Meanwhile, the highest inhibitory activity of ?-glucosidase was shown by E. faecium G.6PR (45%).

scholarly journals Animal Rennets as Sources of Dairy Lactic Acid Bacteria

2014 ◽  
Vol 80 (7) ◽  
pp. 2050-2061 ◽  
Author(s):  
Margherita Cruciata ◽  
Ciro Sannino ◽  
Danilo Ercolini ◽  
Maria L. Scatassa ◽  
Francesca De Filippis ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Streptococcus Thermophilus ◽  
Species Level ◽  
Lactobacillus Delbrueckii ◽  
Rrna Gene ◽  
Microbial Composition ◽  
Content Type

ABSTRACTThe microbial composition of artisan and industrial animal rennet pastes was studied by using both culture-dependent and -independent approaches. Pyrosequencing targeting the 16S rRNA gene allowed to identify 361 operational taxonomic units (OTUs) to the genus/species level. Among lactic acid bacteria (LAB),Streptococcus thermophilusand some lactobacilli, mainlyLactobacillus crispatusandLactobacillus reuteri, were the most abundant species, with differences among the samples. Twelve groups of microorganisms were targeted by viable plate counts revealing a dominance of mesophilic cocci. All rennets were able to acidify ultrahigh-temperature-processed (UHT) milk as shown by pH and total titratable acidity (TTA). Presumptive LAB isolated at the highest dilutions of acidified milks were phenotypically characterized, grouped, differentiated at the strain level by randomly amplified polymorphic DNA (RAPD)-PCR analysis, and subjected to 16S rRNA gene sequencing. Only 18 strains were clearly identified at the species level, asEnterococcus casseliflavus,Enterococcus faecium,Enterococcus faecalis,Enterococcus lactis,Lactobacillus delbrueckii, andStreptococcus thermophilus, while the other strains, all belonging to the genusEnterococcus, could not be allotted into any previously described species. The phylogenetic analysis showed that these strains might represent different unknown species. All strains were evaluated for their dairy technological performances. All isolates produced diacetyl, and 10 of them produced a rapid pH drop in milk, but only 3 isolates were also autolytic. This work showed that animal rennet pastes can be sources of LAB, mainly enterococci, that might contribute to the microbial diversity associated with dairy productions.

A PCR Assay for Detection of Acetic Acid–Tolerant Lactic Acid Bacteria in Acidic Food Products

2004 ◽  
Vol 67 (3) ◽  
pp. 610-615 ◽  
Author(s):  
SHIGERU NAKANO ◽  
ATSUSHI MATSUMURA ◽  
TOSHIHIRO YAMADA
Keyword(s):  
Acetic Acid ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Enrichment Culture ◽  
Rrna Gene ◽  
Bacterial Strains ◽  
Pcr Assay ◽  
Acid Tolerant

A PCR assay for the detection of acetic acid–tolerant lactic acid bacteria in the genera of Lactobacillus and Pediococcus was developed in this study. Primers targeting the bacterial 16S rRNA gene were newly designed and used in this PCR assay. To determine the specificity of the assay, 56 different bacterial strains (of 33 genera), 2 fungi, 3 animals, and 4 plants were tested. Results were positive for most tested bacterial members of 16S rRNA gene–based phylogenetic groups (classi ed in the Lactobacillus casei and Pediococcus group), including Lactobacillus fructivorans, Lactobacillus brevis, Lactobacillus buchneri, Lactobacillus plantarum, and Lactobacillus paracasei. For all other bacterial strains and eukaryote tested, results were negative. Bacterial DNA for PCR was prepared with a simple procedure with the use of Chelex 100 resin from culture after growth in deMan Rogosa Sharpe broth (pH 6.0). To test this PCR assay for the monitoring of the acetic acid–tolerant lactic acid bacteria, L. fructivorans was inoculated into several acidic food as an indicator. Before the PCR, the inoculation of 10 to 50 CFU of bacteria per g of food was followed by a 28-h enrichment culture step, and the PCR assay allowed the detection of bacterial cells. Including the enrichment culture step, the entire PCR detection process can be completed within 30 h.

scholarly journals Molecular characterization of lactic acid bacteria producing edible biofilm isolated from kimchi

2020 ◽  
Vol 21 (3) ◽  
Author(s):  
FADILLA SAPALINA ◽  
Endah Retnaningrum
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Lactobacillus Brevis ◽  
Fermented Food ◽  
Gene Isolation ◽  
Assay Method ◽  
Rrna Gene ◽  
Production Test

Biofilm is a community of microorganisms that interrelated and covered by an extracellular polymer matrix. This biofilm can be produced by Lactic acid bacteria (LAB) which was edible for human and animal.  Therefore, it possible to be applied in the food and health industry. One source of LAB is kimchi, as fermented food from Korea and it has good benefits for health. This research aimed to obtain LAB from kimchi that can produce edible biofilms and to identify LAB producing edible biofilms based on the 16S rRNA gene. Isolation of lactic acid bacteria from kimchi cultured in MRS agar medium containing 1% CaCO3, biofilm production test with biofilm assay method and zeolite as a substrate to adherent cells. For amplification of 16S rRNA gene used primer 27F and 1492R. The isolate of KA2, KA5, KB1, and KC4 isolated from kimchi and could produce biofilms with the highest biofilm formation at 48 hours incubation time. Based on molecular identification with 16S rRNA gene sequencing, the four isolates identified as Lactobacillus brevis species.

scholarly journals Lactobacillus floricola sp. nov., lactic acid bacteria isolated from mountain flowers

2011 ◽  
Vol 61 (6) ◽  
pp. 1356-1359 ◽  
Author(s):  
Shinji Kawasaki ◽  
Kana Kurosawa ◽  
Madoka Miyazaki ◽  
Chisato Yagi ◽  
Yoritaka Kitajima ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Sequence Similarity ◽  
Rrna Gene ◽  
The Novel ◽  
Gene Sequences ◽  
Significant Relationships ◽  
Lactobacillus Kefiri

Five strains (Ryu1-2T, Gon2-9, Ryu4-3, Nog8-1 and Aza1-1) of lactic acid bacteria were isolated from flowers in mountainous areas in Japan, Oze National Park, Iizuna mountain and the Nikko area. The five isolates were found to share almost identical (99.6–100 % similar) 16S rRNA gene sequences and were therefore deemed to belong to the same species. These isolates exhibited low levels of 16S rRNA gene sequence similarity to known lactic acid bacteria; the closest recognized relatives to strain Ryu1-2T were the type strains of Lactobacillus hilgardii (92.8 % similarity), Lactobacillus kefiri (92.7 %), Lactobacillus composti (92.6 %) and Lactobacillus buchneri (92.4 %). Comparative analyses of rpoA and pheS gene sequences demonstrated that the novel isolates did not show significant relationships to other Lactobacillus species. The strains were Gram-stain-positive, catalase-negative and homofermentative. The isolates utilized a narrow range of carbohydrates as sources of carbon and energy, including glucose and fructose. On the basis of phenotypic characteristics and phylogenetic data, these isolates represent a novel species of the genus Lactobacillus, for which the name Lactobacillus floricola sp. nov. is proposed. The type strain is Ryu1-2T ( = NRIC 0774T  = JCM 16512T  = DSM 23037T).

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