16S rDNA Genotyping of Lactic Acid Bacteria Using PCR-RFLP Analysis

Background and Objectives: Lontar (Borassus flabellifer L.) is widely grown in Indonesia and one of its products is palm sap. Palm sap contains a high level of sugar, making it suitable as a medium to increase the lactic acid bacteria (LAB) production of exopolysaccharides (EPS). This study aimed to isolate the EPS-producing LAB from palm sap and evaluate its EPS production. LAB isolation was carried out on MRS agar containing 0.5% CaCO3 . Materials and Methods: The screening and production of EPS were carried out on MRS media supplemented with 10% sucrose. The molecular identification of the selected EPS-producing LAB was based on 16S rDNA. A quantitative analysis of EPS polymer dry mass and total sugar was conducted using one-way ANOVA. Results: In this study, five EPS-producing LABs were found: Fructobacillus fructosus N4, Leuconostoc mesenteroides N5, Leuconostoc mesenteroides N7, Leuconostoc mesenteroides N9, and Fructobacillus fructosus N10. The highest EPS yield in liquid media was 10.997 ± 1.591 g/L by Leuconostoc mesenteroides N7, whereas the lowest was 4.505 ± 0.459 g/L by Fructobacillus fructosus N10. Conclusion: This study found Fructobacillus fructosus strains as EPS producers that have never been reported before.

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Identification and Characterization Indigenous of Lactobacillus spfrom Bovine Rumen Fluidof Slaughterhouse

KnE Life Sciences ◽

10.18502/kls.v3i6.1182 ◽

2017 ◽

Vol 3 (6) ◽

pp. 549

Author(s):

Tri Nurhajati ◽

Koesnoto Soepranianondo ◽

Widya Paramita Lokapirnasari ◽

Adriana Monica Sahidu

Keyword(s):

Lactic Acid ◽

Nucleotide Sequence ◽

Lactic Acid Bacteria ◽

16S Rdna ◽

Biochemical Characterization ◽

Rumen Fluid ◽

Lactobacillus Rhamnosus ◽

Bovine Rumen ◽

Pcr Method

The discovery and characterisation of indigenous lactic acid bacteria (LAB) are important for diversity microbes as candidate probiotic. This research was aimed to identify lactic acid bacteria isolate from isolation process of local bovine rumen fluid from slaughterhouse in Surabaya Indonesia. Genotypic testing was conducted by analyzing 16S rDNA and biochemical identification. DNA of sample isolate was isolated and then amplified in vitro through the PCR method. Determination of nucleotide sequence of 16S rDNA was performed with sequencing method. The result of nucleotide sequence was than compared with GenBank database. The BLAST was then applied to identify the phylogenetic tree. Based on the biochemical characterization and nucleotide sequences, that isolate was identified as Lactobacillus rhamnosus subsp TG15. The result of this research showed that L.rhamnosus subsp TG15 showed viability bacteria in MRSA as control as much as 1.1 x 108 CFU/ml, mean while in MRSA pH 2, L.rhamnosus subsp TG15 showed its viability as much as 9.3 x 106 CFU/ml. Viability of isolate on bile tolerance 0.3% was 2.4 x 107 CFU/ml). Index antagonist bacteria test on S.aureus showed inhibition diameter as much as 2.0 mm and in antagonist test on E.coli as much as 2.5 mm. Based on the result, it could be concluded that this research found a new strain of lactic acid bacteria, L.rhamnosus subsp TG15 and that isolate has ability as the probiotic candidate. Keywords: L.rhamnosus subsp TG15; survival on acidity; bile salts; S.aureus and E.coli

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Identiﬁcation and Characteristics of Lactic Acid Bacteria Isolated from Stylosanthes guianensis sw.

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.361-363.701 ◽

2011 ◽

Vol 361-363 ◽

pp. 701-706 ◽

Cited By ~ 1

Author(s):

Hai Rui Zhai ◽

Han Lin Zhou ◽

Jin Song Yang ◽

Qian Wang ◽

Hai Sheng Tan ◽

...

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

16S Rdna ◽

Enterococcus Faecalis ◽

Acid Production ◽

Lactic Acid Production ◽

Hainan Province ◽

Enterococcus Hirae ◽

Stylosanthes Guianensis ◽

Better Than

Four lactic acid bacteria (LAB) strains named HN78, HN87, HN91 and HN113 were separated fromStylosanthes guianensis SW. of Hainan province, which were identified bacterially using the classical classification and the sequences of 16S rDNA. The results showed that four strains wereEnterococcus hirae,Enterococcus faecalis,Lactococcus lactisandLactobacillus plantarum. The capability of lactic acid production of HN113 and HN91 were better than HN78 and HN87.

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Diversity of Lactic Acid Bacteria in dairy products of Southern Rajasthan

South Asian Journal of Experimental Biology ◽

10.38150/sajeb.3(2).p92-96 ◽

2013 ◽

Vol 3 (2) ◽

pp. 92-96

Author(s):

Nadim Chishty ◽

Srinivasan R ◽

Dinesh K. Kumawat ◽

Yogesh Franklin ◽

Anil Tripathi

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

16S Rdna ◽

Dairy Products ◽

Raw Milk ◽

16S Rdna Gene ◽

Growth Requirements ◽

Rdna Sequencing ◽

Fermented Dairy Products ◽

Fermented Dairy

Lactic acid bacteria (LAB) are a group of gram positive, non spore forming, cocci or rod shaped, catalase negative and fastidious organisms. They are considered as “GRAS” (Generally Recognized As Safe) organisms. A total of 86 bacterial isolates were isolated from different samples of raw milk, buttermilk and curd by using MRS agar and M17 agar. Lactic Acid Bacteria have similar nutritional and growth requirements, it becomes difficult and laborious to identify them by classical methods. Hence, molecular typing was attempted to find the diversity, 16S rDNA gene amplification was done using specific 16S rDNA primers and amplified gene products were digested with different restriction endonuclease enzymes. Further the 16S rDNA sequencing was carried out and the sequences were compared with the available gene sequences in NCBI website by using BLASTn. All isolates obtained from dairy products were accurately identified as Lc. lactis, Lb. plantarum, Leu. mesenteroides, E. durans, Lb. fermentum, Lc. garviae and Lb. casei. The predominant LAB were Lactobacillus casei and Leu. mesenteroides, which were isolated from all sampling sites. The results demonstrate that traditional fermented dairy products from different regions of Rajasthan have complex compositions of LAB species.

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