scholarly journals Molecular identification 16S rRNA gene of active proteolytic lactic acid bacteria (LAB) isolated from kelengkeng (Dimocarpus longan) fruit

2019 ◽  
Vol 20 (8) ◽  
Author(s):  
Habibi Hidayat ◽  
WINARTO HARYADI ◽  
SABIRIN MATSJEH ◽  
TRI JOKO RAHARJO
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Molecular Identification ◽  
Leuconostoc Mesenteroides ◽  
Rrna Gene ◽  
16S Rrna Analysis ◽  
Protease Enzyme ◽  
Dimocarpus Longan

Abstract. Hidayat H, Haryadi W, Matsjeh S, Raharjo TJ. 2019. Molecular identification 16S rRNA gene of active proteolytic lactic acid bacteria (LAB) isolated from kelengkeng (Dimocarpus longan) fruit. Biodiversitas 20: 2222-2228. Various fermentation food involves microorganisms, especially Lactic Acid Bacteria (LAB) which has beneficial properties for fermented food. The present study aims to do identification and characterization of active proteolytic LAB from kelengkeng (Dimocarpus longan), a commonly found fruit in South East Asia. From ten LAB isolates, isolates K7 and K8 appeared as basil shape based on Gram staining and were chosen for further examination. The molecular identification of these selected isolates were investigated from the 16S rRNA analysis using universal primer 27F and 1525R. The K7 and K8 isolates showed resistance ability towards acidic environment (pH 2.0) and were examined for protease enzyme screening resulting 7.5 and 6 to clear zone, which was 17 and 12 mm. The result of the molecular characterization from the two isolates indicated that isolate K7 was identified as Leuconostoc mesenteroides subsp suionicum strain LT-38 while isolate K8 was identified as Leuconostoc mesenteroides strain C305.16.

scholarly journals Molecular identification and phylogenetic analysis of GABA-producing lactic acid bacteria isolated from indigenous dadih of West Sumatera, Indonesia

F1000Research ◽  
2019 ◽  
Vol 7 ◽  
pp. 1663
Author(s):  
Lili Anggraini ◽  
Yetti Marlida ◽  
Wizna Wizna ◽  
Jamsari Jamsari ◽  
Mirzah Mirzah ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Molecular Identification ◽  
Gene Sequence ◽  
Buffalo Milk ◽  
Rrna Gene ◽  
Rrna Gene Sequence

Background: Dadih (fermented buffalo milk) is a traditional Indonesian food originating from West Sumatra province. The fermentation process is carried out by lactic acid bacteria (LAB), which are naturally present in buffalo milk.  Lactic acid bacteria have been reported as one of potential producers of γ-aminobutyric acid (GABA). GABA acts as a neurotransmitter inhibitor of the central nervous system. Methods: In this study, molecular identification and phylogenetic analysis of GABA producing LAB isolated from indigenous dadih of West Sumatera were determined. Identification of the GABA-producing LAB DS15 was based on conventional polymerase chain reaction. 16S rRNA gene sequence analysis was used to identify LAB DS15. Results: PCR of the 16S rRNA gene sequence of LAB DS15 gave an approximately 1400 bp amplicon.  Phylogenetic analysis showed that LAB DS15 was Pediococcus acidilactici, with high similarity of 99% at 100% query coverage to Pediococcus acidilactici strain DSM 20284. Conclusions: It can be concluded that GABA producing LAB isolated from indigenous dadih was Pediococcus acidilactici.

scholarly journals Molecular identification and phylogenetic analysis of GABA-producing lactic acid bacteria isolated from indigenous dadih of West Sumatera, Indonesia

F1000Research ◽  
2019 ◽  
Vol 7 ◽  
pp. 1663
Author(s):  
Lili Anggraini ◽  
Yetti Marlida ◽  
Wizna Wizna ◽  
Jamsari Jamsari ◽  
Mirzah Mirzah ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Molecular Identification ◽  
Gene Sequence ◽  
Buffalo Milk ◽  
Rrna Gene ◽  
Rrna Gene Sequence

Background: Dadih (fermented buffalo milk) is a traditional Indonesian food originating from West Sumatra province. The fermentation process is carried out by lactic acid bacteria (LAB), which are naturally present in buffalo milk.  Lactic acid bacteria have been reported as one of potential producers of γ-aminobutyric acid (GABA). GABA acts as a neurotransmitter inhibitor of the central nervous system. Methods: In this study, molecular identification and phylogenetic analysis of GABA producing LAB isolated from indigenous dadih of West Sumatera were determined. Identification of the GABA-producing LAB DS15 was based on conventional polymerase chain reaction. 16S rRNA gene sequence analysis was used to identify LAB DS15. Results: PCR of the 16S rRNA gene sequence of LAB DS15 gave an approximately 1400 bp amplicon.  Phylogenetic analysis showed that LAB DS15 was Pediococcus acidilactici, with high similarity of 99% at 100% query coverage to Pediococcus acidilactici strain DSM 20284. Conclusions: It can be concluded that GABA producing LAB isolated from indigenous dadih was Pediococcus acidilactici.

scholarly journals Molecular identification and phylogenetic analysis of GABA-producing lactic acid bacteria isolated from indigenous dadih of West Sumatera, Indonesia

F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 1663
Author(s):  
Lili Anggraini ◽  
Yetti Marlida ◽  
Wizna Wizna ◽  
Jamsari Jamsari ◽  
Mirzah Mirzah ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Molecular Identification ◽  
Gene Sequence ◽  
Buffalo Milk ◽  
Rrna Gene ◽  
Rrna Gene Sequence

Background: Dadih (fermented buffalo milk) is a traditional Indonesian food originating from West Sumatra province. The fermentation process is carried out by lactic acid bacteria (LAB), which are naturally present in buffalo milk.  Lactic acid bacteria have been reported as one of potential producers of γ-aminobutyric acid (GABA). GABA acts as a neurotransmitter inhibitor of the central nervous system. Methods: In this study, molecular identification and phylogenetic analysis of GABA producing LAB isolated from indigenous dadih of West Sumatera were determined. Identification of the GABA-producing LAB DS15 was based on conventional polymerase chain reaction. 16S rRNA gene sequence analysis was used to identify LAB DS15. Results: PCR of the 16S rRNA gene sequence of LAB DS15 gave an approximately 1400 bp amplicon.  Phylogenetic analysis showed that LAB DS15 was Pediococcus acidilactici, with high similarity of 99% at 100% query coverage to Pediococcus acidilactici strain DSM 20284. Conclusions: It can be concluded that GABA producing LAB isolated from indigenous dadih was Pediococcus acidilactici.

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.

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