scholarly journals Streptomyces flavogriseusHS1: Isolation and Characterization of Extracellular Proteases and Their Compatibility with Laundry Detergents

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Sofiane Ghorbel ◽  
Maher Kammoun ◽  
Hala Soltana ◽  
Moncef Nasri ◽  
Noomen Hmidet

The present study describes the isolation of a new protease producingStreptomycesstrain HS1 and the biochemical characterization of the secreted proteases. By sequencing of its noted 16S rDNA, HS1 strain was found to have a 100% identity withStreptomyces flavogriseus. The highest protease production was found using FermII media. In these conditions maximum protease production (99 U/mL) was obtained after 96 h incubation at 30°C and 150 rpm. HS1 strain produced at least five proteases as revealed by zymogram technique. The enzyme preparation exhibited activity over a broad range of pH (5–11) and temperature (25–70°C). Optimum activity was observed at a pH of 7.0 and a temperature of 50°C. Proteolytic activity was significantly unaffected by Ca2+and Mg2+. EDTA and PMSF highly decreased the original activity. The crude extracellular proteases showed high stability when used as a detergent additive. These properties offer an interesting potential for enzymatic hydrolysis at the industrial level.

Certain microorganisms especially bacteria and fungi are able to use xenobiotic organic compounds as their carbon and nitrogen source for metabolism. Flower farms around lake Naivasha basin uses several agrochemicals especially pesticides to control pests and improve flower production. The aim of this study was to isolate and characterize morphologically and biochemically the main bacterial species that are able to grow and tolerate the pesticide contaminated farm soils. Soil samples were collected from randomly selected five greenhouses from each five flower farms namely Crescent, Elsamere, Karuturi, Malewa and Sewage farms around Lake Naivasha basin. The collected samples were processed for bacterial isolation using the nutrient agar, mac’ Conkey agar, blood agar, Luria-Bertani and Minimum Salt Media nutrient media. The conventional methods of swabbing and streaking were used. Pure colonies of isolates organisms were identified and characterized using standard microbiological technique. Morphological, cultural and biochemical characterization of bacterial species isolated from the flower farm soil samples identified mainly Pseudomonas auriginosa, Escherichia coli, Rhodococcus erythropolis and Bacillus subtilis species. Bacterial growth in pesticide consortia was quantified by monitoring colony growth of the species in liquid culture over time. The viable cell counts were determined turbidimetrically at O.D696nm. All the isolated bacterial species were able to grow in flower farm soil contaminated with organochloride and organophosphate pesticide residues. B. subtilis recorded the highest growth at 1.77±0.07 O.D696nm in pesticide mixture consortia. There was lower growth in organochloride pesticide consortia as compared to organophosphate pesticide consortia.


2021 ◽  
Vol 16 (7) ◽  
pp. 122-129
Author(s):  
Neha Sharma ◽  
Shuchi Kaushik ◽  
Rajesh Singh Tomar

Microbial glutaminase and urease have demonstrated their benefits in various fields like medicinal, pharmaceutical and biotechnological industries. Keeping this viewpoint, the aim of the present study was the isolation and characterization of extracellular enzyme-producing bacteria from soil samples collected from different regions of Gwalior (M.P.). The isolated bacterial cultures were processed by serial dilution method and maintained on nutrient agar medium following standard microbiological laboratory practices for maintenance and preservation of bacteria. We screened out three enzyme producing strains of Salmonella sp., Proteus vulgaris and Bacillus subtilis. The screening was based on biochemical testing and enzyme assays. To accomplish this work, we used differential as well as selective media. All the selected isolates were able to produce enzymes like L-Glutaminase and Urease with different specific enzymatic activity. These bacterial isolates were not reported to show any type of allergenicity when their sequences were checked by bioinformatics tool Algpred. So, these bacterial isolates can be considered as an alternative source for the production of enzymes and can be used for largescale production of enzymes at the industrial level.


2019 ◽  
Vol 295 (6) ◽  
pp. 1598-1612 ◽  
Author(s):  
Ivette M. Menéndez-Perdomo ◽  
Peter J. Facchini

Benzylisoquinoline alkaloids (BIAs) are a major class of plant metabolites with many pharmacological benefits. Sacred lotus (Nelumbo nucifera) is an ancient aquatic plant of medicinal value because of antiviral and immunomodulatory activities linked to its constituent BIAs. Although more than 30 BIAs belonging to the 1-benzylisoquinoline, aporphine, and bisbenzylisoquinoline structural subclasses and displaying a predominant R-enantiomeric conformation have been isolated from N. nucifera, its BIA biosynthetic genes and enzymes remain unknown. Herein, we report the isolation and biochemical characterization of two O-methyltransferases (OMTs) involved in BIA biosynthesis in sacred lotus. Five homologous genes, designated NnOMT1–5 and encoding polypeptides sharing >40% amino acid sequence identity, were expressed in Escherichia coli. Functional characterization of the purified recombinant proteins revealed that NnOMT1 is a regiospecific 1-benzylisoquinoline 6-O-methyltransferase (6OMT) accepting both R- and S-substrates, whereas NnOMT5 is mainly a 7-O-methyltransferase (7OMT), with relatively minor 6OMT activity and a strong stereospecific preference for S-enantiomers. Available aporphines were not accepted as substrates by either enzyme, suggesting that O-methylation precedes BIA formation from 1-benzylisoquinoline intermediates. Km values for NnOMT1 and NnOMT5 were 20 and 13 μm for (R,S)-norcoclaurine and (S)-N-methylcoclaurine, respectively, similar to those for OMTs from other BIA-producing plants. Organ-based correlations of alkaloid content, OMT activity in crude extracts, and OMT gene expression supported physiological roles for NnOMT1 and NnOMT5 in BIA metabolism, occurring primarily in young leaves and embryos of sacred lotus. In summary, our work identifies two OMTs involved in BIA metabolism in the medicinal plant N. nucifera.


2003 ◽  
Vol 93 (4) ◽  
pp. 485-492 ◽  
Author(s):  
Elena G. Biosca ◽  
Raquel González ◽  
María José López-López ◽  
Santiago Soria ◽  
Carmina Montón ◽  
...  

The drippy nut disease of oak was first described in California in 1967 and, since then, the causal agent has not been reported in any other area. This study describes for the first time in Europe the isolation of Brenneria (Erwinia) quercina from bark canker in addition to drippy bud and drippy nut in Quercus ilex and Q. pyrenaica. The bark canker and drippy bud symptoms were not previously described as caused by this bacterium. No fungal pathogens were associated with any of the symptoms. Physiological and biochemical characterization identified the pathogenic isolates from Spain as belonging to B. quercina, similar to the reference strain CFBP 1266. Fatty acid profiles of the Spanish isolates also were similar to the strain of B. quercina from California. Serological analysis by indirect immunofluorescence and enzyme-linked immunosorbent assay using polyclonal antisera against the reference strain of B. quercina and one Spanish oak isolate revealed some antigenic heterogeneity between isolates of different origins. Pathogenicity tests demonstrated that the Spanish isolates were able to reproduce internal symptoms of necrosis and acorn exudation in Q. ilex and Q. pyrenaica and suggest that B. quercina may be associated, among other causes, with the oak decline syndrome affecting Spanish oak forests.


1984 ◽  
Vol 218 (2) ◽  
pp. 591-599 ◽  
Author(s):  
T Saermark ◽  
P M Jones ◽  
I C A F Robinson

We have developed small-scale methods for the isolation and biochemical characterization of subcellular fractions from single guinea-pig posterior-pituitary glands. Secretory vesicles and coated microvesicles produced in this way were of similar purity to those isolated from large amounts of tissue by conventional ultracentrifugation. [35S]Cysteine injected into the hypothalamus was found in the soluble contents of secretory vesicles isolated from the neural lobes 24 h later. High-pressure liquid-chromatographic analysis revealed that the radiolabel was incorporated into the expected neurosecretory products (oxytocin, vasopressin and neurophysin) and also into a biosynthetic intermediate in the vasopressin system. The membranes of secretory vesicles were labelled with [3H]choline 24 h after its hypothalamic injection. Little or no [3H]choline could be demonstrated in coated microvesicles at this time, although these structures were labelled 5 days after injection. Stimulating hormone secretion by chronic dehydration produced a significant fall in [3H]choline content of the secretory-vesicle membranes without any transfer of label into coated microvesicles, suggesting that coated microvesicles are not involved in membrane retrieval in the neurohypophysis.


2020 ◽  
Vol 11 (1) ◽  
pp. 818-825
Author(s):  
Kolli Guna Ranjan ◽  
Girija Sankar G ◽  
Satyanarayana Raju D V V

Commercial probiotics have bacteria which offer scope for specific treatment as a probiotic against diarrhoea, boosting immune response and relieving stress. Research in probiotics and aquaculture has been done to promote sustainable aquaculture. Studies on microbial ecology in aquaculture and their benefits on human systems are to be assessed. Hence, the present study was aimed at the isolation and characterization of these bacteria obtained from various commercial probiotics. The commercial probiotic samples were collected for isolation using MRS agar. The colonies were selected on the basis of colonial morphology.  Isolates were put through cell morphology, physiology and different biochemical tests. Probiotic sample confirmation was done using 16srRNA molecular technique. MEGA7 was used to conduct phylogenetic evolutionary analysis and tabulate a distance matrix. In results, the isolates manifested non-identical growth patterns at dissimilar conc. of NaCl (2.0,4.0 and 6.5) oxygen and at different temperatures (150C, 300C and 450C). On the basis of sugar utilization, physiological testing, biochemical characterization and genetic identification tests, all isolates were established to different species of Lactobacillus rhamnosus, Pediococcus acidilactici and Lactobacillus Plantarum. A systematic protocol was done to identify, characterize probiotic samples and identify them by genetic analysis. Probiotic use is carefully assessed by regulations


Sign in / Sign up

Export Citation Format

Share Document