scholarly journals Isolation and Characterization of L-Glutaminase producing Bacteria

2020 ◽  
Author(s):  
Rabia Saleem ◽  
Safia Ahmed

AbstractBeing a significant protein L-glutaminases discovers potential applications in various divisions running from nourishment industry to restorative and cure. It is generally disseminated in microbes, actinomycetes, yeast and organisms. Glutaminase is the principal enzyme that changes glutamine to glutamate. The samples were gathered from soil of Taxila, Wah Cantt and Quetta, Pakistan for the isolation of glutaminase producing bacteria. After primary screening, subordinate screening was done which includes multiple testification such as purification, observation of morphological characters and biochemical testing of bacterial strains along with 16S rRNA sequence homology testing. Five bacterial strains were selected showing glutaminase positive test in screening, enzyme production via fermentation and enzymatic and protein assays. Taxonomical characterization of the isolates identified them as Bacillus subtilis U1, Achromobacter xylosoxidans G1, Bacillus subtilis Q2, Stenotrophomonas maltophilia U3 and Alcaligenes faecalis S3. The optimization of different effectors such as incubation time, inducers, carbon source, pH, and nitrogen source were also put under consideration. There was slight difference among incubation of bacterial culture, overall, 36 hours of incubation time was the best for glutaminase production by all the strains. Optimal pH was around 9 in Achromobacter xylosoxidans G1 and Alcaligenes faecalis S3, pH 6 in Bacillus subtilis U1, pH 8 in Stenotrophomonas maltophilia U3, pH 6-8 in Bacillus subtilis Q2. Best glutaminase production was obtained at 37°C by Bacillus subtilis U1and Bacillus subtilis Q2, 30°C for Achromobacter xylosoxidans G1, Stenotrophomonas maltophilia U3 and 25°C by Alcaligenes faecalis S3. The carbon sources put fluctuated effects on activity of enzyme in such a way that glucose was the best carbon source for Bacillus subtilis U1and Bacillus subtilis Q2, Sorbitol for Achromobacter xylosoxidans G1 and Alcaligenes faecalis S3 while xylose was the best for Stenotrophomonas maltophilia U3. Yeast extract and Trypton were among good nitrogen sources for Achromobacter xylosoxidans G1 and of Bacillus subtilis U1 respectively. Glutamine was the best inducer for Bacillus subtilis Q2, Alcaligenes faecalis S3 and Stenotrophomonas maltophilia U3, while lysine for Achromobacter xylosoxidans G1 and glycine act as good inducer in case of Bacillus subtilis U1. After implementation of optimal conditions microbial L-glutaminase production can be achieved and the bacterial isolates have a great potential for production of glutaminase enzyme and their applications.

2015 ◽  
Vol 77 (31) ◽  
Author(s):  
Suganthi Thevarajoo ◽  
Chitra Selvaratnam ◽  
Kian Mau Goh ◽  
Fazilah Abd. Manan ◽  
Zaharah Ibrahim ◽  
...  

Marine environment remained as largely unexplored source for researchers to discover marine microorganisms with novel properties. This study aims to isolate marine bacteria from the seashore of Desaru, Malaysia. Totally, six bacterial strains were successfully obtained and were identified by complete 16S rRNA sequencing. The characterizations of bacterial strains were performed based on morphological tests, Gram-staining, biochemical tests, and antibiotic sensitivity. The 16S rRNA sequence of D-2, D-4, D-7, D-15, D-31, and D-33 revealed a high identity of 97 to 99% with taxa belong to genera of Pseudomonas, Marinomonas, Exiquobacterium, Micrococcus, Pseudoalteromonas, and Shewanella respectively. Strain D-31 exhibited higher tolerance towards antibiotics kanamycin, ampicillin, and erythromycin while the growth of other strains were retarded by at least two of these antibiotics. We further characterized strain D-4 and D-31 that belonged to Marinomonas sp. and Pseudoalteromonas sp.. Both genera are interesting as earlier researchers have discovered new antibacterial substances, industrial enzymes and unique secondary metabolites.


Converting spent mushroom substrates into organic fertilizer helps to tackle the problem of pollution in edible canna starch processing villages and adds new value to the production chain of edible canna. To successfully turn the spent substrates into compost, there is certainly an indispensable role for cellulolytic microorganisms, in which Bacillus strains are always important. Several bacterial strains have been isolated from spent edible canna substrate after cultivation of monkey head mushroom in this study. Among isolated strains, the strain NDK5 has been selected exhibiting the highest cellulolytic activities with solubilization indexes of 6.14 and 18.3 mm for the ratio between the halo zone diameters and the colony diameters in the point cultivation method (SIratio) and the offset between the halo zone diameters and the agar hole diameters (SIoffset), respectively. The highest CMCase activity was 4.29 ± 0.071 U/ml. Morphological, physiological, biochemical, and 16S rRNA sequence analyses (100% homology with B. amyloliquefaciens sp. plantarum FZB42) were further carried out for the selected strain, leading to the identification of the strain as B. amyloliquefaciens sp. plantarum NDK5 strain. In addition, NDK5 was proved to have a capacity for synthesizing indole-3-acetic acid, a plant growth hormone, on an L-tryptophan-containing medium. Trial incubation of spent mushroom edible canna-substrate with the strain NDK5 showed increases in several quality criteria of the waste after 20 days of incubation, that meet the standard criteria for bio-organic fertilizer according to TCVN 7185:2002.


2021 ◽  
Vol 9 (2) ◽  
pp. 7-12
Author(s):  
Abdulrahman Abdulhamid Arabo ◽  
Raji Arabi Bamanga ◽  
Mujiburrahman Fadilu ◽  
Musa Abubakar ◽  
Fatima Abdullahi Shehu ◽  
...  

This study aimed to isolate and identify biosurfactant producing and diesel alkanes degrading bacteria. For this reason, bacteria isolated from the diesel contaminated site were screened for their potential to produce biosurfactants and degrade diesel alkanes. Primary selection of diesel degraders was carried out by using conventional enrichment culture technique where 12 bacterial strains were isolated based on their ability to grow on minimal media supplemented with diesel as sole carbon source, which was followed by qualitative screening methods for potential biosurfactant production. Isolate B11 was the only candidate that shows positive signs for drop collapse, foaming, haemolytic test, oil displacement of more than 22 ± 0.05 mm, and emulsification (E24) of 14 ± 0.30%. The effect of various culture parameters (incubation time, diesel concentration, nitrogen source, pH and temperature) on biodegradation of diesel was evaluated. The optimum incubation time was confirmed to be 120 days for isolates B11, the optimum PH was confirmed as 8.0 for the isolate, Similarly, the optimum temperature was confirmed as 35oC. In addition, diesel oil was used as the sole carbon source for the isolates. The favourable diesel concentration was 12.5 % (v/v) for the isolate. The isolate has shown degradative ability towards Tridecane (C13), dodecane, 2, 6, 10-trimethyl- (C15), Tetradecane (C14), 2,6,10-Trimethyltridecane (C16), Pentadecane (C15). It degraded between 0.27% - 9.65% individual diesel oil alkanes. The strain has exhibited the potential of degrading diesel oil n-alkanes and was identified as Alcaligenes species strain B11 (MZ027604) using the 16S rRNA sequencing.


Respuestas ◽  
2020 ◽  
Vol 25 (1) ◽  
pp. 127-137
Author(s):  
Liliana Yanet Suárez-Contreras ◽  
Luz Francy Yañez-Meneses

Bacterial identification is carried out by conventional methods based on phenotypic characteristics, since their implementation and costs are more easily accessible. However, molecular identification allows us to know the true identity of the genus and species. The molecular identification of 24 bacterial strains preserved in the Strain Bank of the University Francisco de Paula Santander, Campos Eliseos Experimental Center, identified under macroscopic and microscopic phenotypic criteria, was carried out. Initially, the strains preserved in saline solution were reactivated and characterized macro- and microscopically, then DNA extraction was performed and PCR was done to amplify the 16S rRNA region allowing access to the DNA sequence of interest; the samples were sent to be sequenced and through bioinformatic tools the identity of each bacterium was known. The strains: BLB003, BLB009, BLB011, BLB012, BLB014, BLB016, BLB018, BLB022, BLB023, BLB024, BLB033, were identified as Bacillus cereus; BLB010 as Bacillus thurigiensis; while BLB030, BLB031, BLB032, as Bacillus pumilus; BLB020 as Bacillus amyloliquefaciens; BLB001, BLB004, BLB007, and BLB037, formed the group of Bacillus subtilis; and it is possible that there are divergent ramifications between species of Bacillus in phylogenetic trees. Another grouping that was observed in the phylogenetic tree are the strains BLB019 and BLB029 that correspond to Achromobacter xylosoxidans and Alcaligenes faecalis respectively. Also another group BLB013 and BLB017, were identified as Stenotrophomonas maltophilia. It is important to take into account that sometimes 16S rRNA presents a low discrimination capacity for some genera and species due to recent divergences, it is necessary to complement the identification with the study of other genes.


2002 ◽  
Vol 68 (9) ◽  
pp. 4390-4398 ◽  
Author(s):  
S. A. F. T. van Hijum ◽  
G. H. van Geel-Schutten ◽  
H. Rahaoui ◽  
M. J. E. C. van der Maarel ◽  
L. Dijkhuizen

ABSTRACT Fructosyltransferase (FTF) enzymes produce fructose polymers (fructans) from sucrose. Here, we report the isolation and characterization of an FTF-encoding gene from Lactobacillus reuteri strain 121. A C-terminally truncated version of the ftf gene was successfully expressed in Escherichia coli. When incubated with sucrose, the purified recombinant FTF enzyme produced large amounts of fructo-oligosaccharides (FOS) with β-(2→1)-linked fructosyl units, plus a high-molecular-weight fructan polymer (>107) with β-(2→1) linkages (an inulin). FOS, but not inulin, was found in supernatants of L. reuteri strain 121 cultures grown on medium containing sucrose. Bacterial inulin production has been reported for only Streptococcus mutans strains. FOS production has been reported for a few bacterial strains. This paper reports the first-time isolation and molecular characterization of (i) a Lactobacillus ftf gene, (ii) an inulosucrase associated with a generally regarded as safe bacterium, (iii) an FTF enzyme synthesizing both a high molecular weight inulin and FOS, and (iv) an FTF protein containing a cell wall-anchoring LPXTG motif. The biological relevance and potential health benefits of an inulosucrase associated with an L. reuteri strain remain to be established.


Extremophiles ◽  
2002 ◽  
Vol 6 (6) ◽  
pp. 499-506 ◽  
Author(s):  
Niran Roongsawang ◽  
Jiraporn Thaniyavarn ◽  
Suthep Thaniyavarn ◽  
Takayuki Kameyama ◽  
Mitsuru Haruki ◽  
...  

2021 ◽  
Vol 16 (8) ◽  
pp. 110-117
Author(s):  
Kannan Abhirami ◽  
K. Jayakumar

Phosphorous is considered as a major parameter for crop yield. Its availability to plant is independent of its abundance. For the plants to utilize phosphorous, it is to be converted to absorbable form. Here, the part rendered by phosphate solubilizing bacteria is significant for it plays a crucial role in the formation of plant usable phosphate from organic forms. In the present work, an effort had been made to isolate and identify phosphate solubilising bacterial isolate from the rhizhospheric soils of various plants in Ponthenpuzha forest. One of the isolate from Cymbopogon citrates responded positively to Pikovskaya’s medium by producing a halo zone during in vitro culture. Colony features and 16S rRNA sequence analysis identified the isolate as Burkholderia sps. We have reported the presence of genus Burkholderia in the rhizospheric zone of Cymbopogon citratus. Further studies are warranted for species level identification of the isolate.


2013 ◽  
Vol 33 (2) ◽  
pp. 295-303 ◽  
Author(s):  
Cintia Anabela Mazzucotelli ◽  
Alejandra Graciela Ponce ◽  
Catalina Elena Kotlar ◽  
María del Rosario Moreira

Nutrients ◽  
2018 ◽  
Vol 10 (11) ◽  
pp. 1684 ◽  
Author(s):  
Larissa Celiberto ◽  
Roseli Pinto ◽  
Elizeu Rossi ◽  
Bruce Vallance ◽  
Daniela Cavallini

Modulation of the gut microbiota through the use of probiotics has been widely used to treat or prevent several intestinal diseases. However, inconsistent results have compromised the efficacy of this approach, especially in severe conditions such as inflammatory bowel disease (IBD). The purpose of our study was to develop a personalized probiotic strategy and assess its efficacy in a murine model of intestinal inflammation. Commensal bacterial strains were isolated from the feces of healthy mice and then administered back to the host as a personalized treatment in dextran sodium sulfate (DSS)-induced colitis. Colonic tissues were collected for histological analysis and to investigate inflammatory markers such as Il-1β, Il-6, TGF-β, and Il-10, and the enzyme myeloperoxidase as a neutrophil marker. The group that received the personalized probiotic showed reduced susceptibility to DSS-colitis as compared to a commercial probiotic. This protection was characterized by a lower disease activity index and reduced histopathological damage in the colon. Moreover, the personalized probiotic was more effective in modulating the host immune response, leading to decreased Il-1β and Il-6 and increased TGF-β and Il-10 expression. In conclusion, our study suggests that personalized probiotics may possess an advantage over commercial probiotics in treating dysbiotic-related conditions, possibly because they are derived directly from the host’s own microbiota.


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