scholarly journals Analysis of 16S rRNA gene lactic acid bacteria (LAB) isolate from Markisa fruit (Passiflora sp.) as a producer of protease enzyme and probiotics

2017 ◽  
Author(s):  
Habibi Hidayat
2019 ◽  
Vol 20 (8) ◽  
Author(s):  
Habibi Hidayat ◽  
WINARTO HARYADI ◽  
SABIRIN MATSJEH ◽  
TRI JOKO RAHARJO

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.


2014 ◽  
Vol 80 (7) ◽  
pp. 2050-2061 ◽  
Author(s):  
Margherita Cruciata ◽  
Ciro Sannino ◽  
Danilo Ercolini ◽  
Maria L. Scatassa ◽  
Francesca De Filippis ◽  
...  

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.


2004 ◽  
Vol 67 (3) ◽  
pp. 610-615 ◽  
Author(s):  
SHIGERU NAKANO ◽  
ATSUSHI MATSUMURA ◽  
TOSHIHIRO YAMADA

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.


2020 ◽  
Vol 21 (3) ◽  
Author(s):  
FADILLA SAPALINA ◽  
Endah Retnaningrum

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.


2011 ◽  
Vol 61 (6) ◽  
pp. 1356-1359 ◽  
Author(s):  
Shinji Kawasaki ◽  
Kana Kurosawa ◽  
Madoka Miyazaki ◽  
Chisato Yagi ◽  
Yoritaka Kitajima ◽  
...  

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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