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402 result(s) for "Subramanian, B. R."
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Degradation insight of organophosphate pesticide chlorpyrifos through novel intermediate 2,6-dihydroxypyridine by Arthrobacter sp. HM01
Organophosphates (OPs) are hazardous pesticides, but an indispensable part of modern agriculture; collaterally contaminating agricultural soil and surrounding water. They have raised serious food safety and environmental toxicity that adversely affect the terrestrial and aquatic ecosystems and therefore, it become essential to develop a rapid bioremediation technique for restoring the pristine environment. A newly OPs degrading Arthrobacter sp. HM01 was isolated from pesticide-contaminated soil and identified by a ribotyping (16S rRNA) method. Genus Arthrobacter has not been previously reported in chlorpyrifos (CP) degradation, which shows 99% CP (100 mg L−1) degradation within 10 h in mMSM medium and also shows tolerance to a high concentration (1000 mg L−1) of CP. HM01 utilized a broad range of OPs pesticides and other aromatic pollutants including intermediates of CP degradation as sole carbon sources. The maximum CP degradation was obtained at pH 7 and 32 °C. During the degradation, a newly identified intermediate 2,6-dihydroxypyridine was detected through TLC/HPLC/LCMS analysis and a putative pathway was proposed for its degradation. The study also revealed that the organophosphate hydrolase (opdH) gene was responsible for CP degradation, and the opdH-enzyme was located intracellularly. The opdH enzyme was characterized from cell free extract for its optimum pH and temperature requirement, which was 7.0 and 50 °C, respectively. Thus, the results revealed the true potential of HM01 for OPs-bioremediation. Moreover, the strain HM01 showed the fastest rate of CP degradation, among the reported Arthrobacter sp.
Lactiplantibacillus plantarum PGB02 Improved Serum Cholesterol Profile by Tweaking Genes Involved in Cholesterol Homeostasis in Male Swiss Albino Mice
The effect of Lactiplantibacillus plantarum PGB02 isolated from buttermilk on serum cholesterol profile of normal and hypercholesterolemic mice was evaluated. Further changes in the expression of mice genes were determined. The hypercholesterolemia was induced in experimental mice by feeding high cholesterol and fat diet. Serum cholesterol parameters, physical parameters, cholic acid excretion, and cholesterol metabolism related gene expression analysis was carried out. L. plantarum PGB02 efficiently reduced total cholesterol, triglycerides, and LDL-cholesterol and improved HDL-cholesterol in hypercholesterolaemic mice. Body weight was reduced and fecal cholic acid increased in probiotic treatment groups. Gene expression analysis revealed that L. plantarum PGB02 up-regulated the expression of LDL receptors, CYP7A1, ABCA1, ABCG5, ABCG8, and down-regulated the expression of FXR and NPC1L1 genes. Summarizing the mechanism, L. plantarum PGB02 improved hypercholesterolemia by increasing bile acid synthesis and excretion, reducing exogeneous cholesterol absorption from the intestine, and increased LDL clearance through upregulation of LDL-receptors. The present study has given insight into the mechanism of serum cholesterol reduction by bile salt hydrolase positive L. plantarum PGB02 in mice. L. plantarum PGB02 reduced the serum cholesterol level through increased bile acid synthesis and deconjugation and reduced absorption of cholesterol in the intestine. Isolate PGB02 shown cholesterol removal potential as good as statin.
Effects of substrate binding site residue substitutions of xynA from Bacillus amyloliquefaciens on substrate specificity
Background The aromatic residues of xylanase enzyme, W187, Y124, W144, Y128 and W63 of substrate binding pocket from Bacillus amyloliquefaciens were investigated for their role in substrate binding by homology modelling and sequence analysis. These residues are highly conserved and play an important role in substrate binding through steric hindrance. The substitution of these residues with alanine allows the enzyme to accommodate nonspecific substrates. Results Wild type and mutated genes were cloned and overexpressed in BL21. Optimum pH and temperature of rBAxn exhibited pH 9.0 and 50 °C respectively and it was stable up to 215 h. Along with the physical properties of rBAxn, kinetic parameters ( K m 19.34 ± 0.72 mg/ml; k cat 6449.12 ± 155.37 min − 1 and k cat /K m 333.83 ± 6.78 ml min − 1  mg − 1 ) were also compared with engineered enzymes. Out of five mutations, W63A, Y128A and W144A lost almost 90% activity and Y124A and W187A retained almost 40–45% xylanase activity. Conclusions The site-specific single mutation, led to alteration in substrate specificity from xylan to CMC while in case of double mutant the substrate specificity was altered from xylan to CMC, FP and avicel, indicating the role of aromatic residues on substrate binding, catalytic process and overall catalytic efficiency.
Investigating the Effect of Pipecolic Acid on Specialized Metabolites Involved in Tomato Plant Defense Mechanisms Against Ralstonia solanacearum Wilt Pathogens
The role of pipecolic acid (Pip) in plant immune responses, particularly against bacterial wilt pathogens, is significant. This research aimed to understand the interaction between plant defense-responsive enzymes and Pip by analyzing methanolic extracts from different treatments of tolerant (GAT5) and susceptible (GT2) tomato cultivars. LC-MS analysis demonstrated that the foliar application of Pip significantly influenced tomato metabolites, especially in bacterial wilt-infected plants, with a more pronounced effect in tolerant varieties. Principal component analysis (PCA) revealed that Pip-treated plants of tolerant varieties exhibited better coordinated metabolome profiles than those of susceptible varieties. Notable variations were observed in the levels of specialized metabolites, such as salicylic acid (SA), N-hydroxy pipecolic acid (NHP), and Pip, which are essential for producing defense compounds. Molecular docking studies further explored Pip’s interactions with key plant enzymes involved in defense mechanisms and showed that Pip acts as an effective organic inducer of systemic acquired resistance (SAR). These findings highlight Pip’s potential as a natural agent for enhancing plant tolerance to pathogens, offering promising implications for agricultural practices and improving crop resilience against diseases. This study enhances our understanding of Pip’s role in plant defense and provides a foundation for developing Pip-based strategies for sustainable agriculture.
Influence of net ecosystem metabolism in transferring riverine organic carbon to atmospheric CO₂ in a tropical coastal lagoon (Chilka Lake, India)
Studies on biogeochemical cycling of carbon in the Chilka Lake, Asia's largest brackish lagoon on the east coast of India, revealed, for the first time, strong seasonal and spatial variability associated with salinity distribution. The lake was studied twice during May 2005 (premonsoon) and August 2005 (monsoon). It exchanges waters with the sea (Bay of Bengal) and several rivers open into the lake. The lake showed contrasting levels of dissolved inorganic carbon (DIC) and organic carbon (DOC) in different seasons; DIC was higher by ~22% and DOC was lower by ~36% in premonsoon than in monsoon due to seasonal variations in their supply from rivers and in situ production/mineralisation. The DIC/DOC ratios in the lake during monsoon were influenced by physical mixing of end member water masses and by intense respiration of organic carbon. A strong relationship between excess DIC and apparent oxygen utilisation showed significant control of biological processes over CO₂ production in the lake. Surface partial pressure of CO₂ (pCO₂), calculated using pH-DIC couple according to Cai and Wang (Limnol and Oceanogr 43:657-668, 1998), exhibited discernable gradients during monsoon through northern (1,033-6,522 μatm), central (391-2,573 μatm) and southern (102-718 μatm) lake. The distribution pattern of pCO₂ in the lake seems to be governed by pCO₂ levels in rivers and their discharge rates, which were several folds higher during monsoon than premonsoon. The net CO₂ efflux, based on gas transfer velocity parameterisation of Borges et al. (Limnol and Oceanogr 49(5):1630-1641, 2004), from entire lake during monsoon (141 mmolC m⁻² d⁻¹ equivalent to 2.64 GgC d⁻¹ at basin scale) was higher by 44 times than during premonsoon (9.8 mmolC m⁻² d⁻¹ [almost equal to] 0.06 GgC d⁻¹). 15% of CO₂ efflux from lake in monsoon was contributed by its supply from rivers and the rest was contributed by in situ heterotrophic activity. Based on oxygen and total carbon mass balance, net ecosystem production (NEP) of lake (-308 mmolC m⁻² d⁻¹ [almost equal to] -3.77 GgC d⁻¹) was found to be almost in consistent with the total riverine organic carbon trapped in the lake (229 mmolC m⁻² d⁻¹ [almost equal to] 2.80 GgC d⁻¹) suggesting that the strong heterotrophy in the lake is mainly responsible for elevated fluxes of CO₂ during monsoon. Further, the pelagic net community production represented 92% of NEP and benthic compartment plays only a minor role. This suggests that Chilka lake is an important region in biological transformation of organic carbon to inorganic carbon and its export to the atmosphere.
Influence of nutrient input on the trophic state of a tropical brackish water lagoon
Ecosystem level changes in water quality and biotic communities in coastal lagoons have been associated with intensification of anthropogenic pressures. In light of incipient changes in Asia’s largest brackish water lagoon (Chilika, India), an examination of different dissolved nutrients distribution and phytoplankton biomass, was conducted through seasonal water quality monitoring in the year 2011. The lagoon showed both spatial and temporal variation in nutrient concentration, mostly altered by freshwater input, regulated the chlorophyll distribution as well. Dissolved inorganic N:P ratio in the lagoon showed nitrogen limitation in May and December, 2011. Chlorophyll in the lagoon varied between 3.38 and 17.66 mg m −3 . Spatially, northern part of the lagoon showed higher values of DIN and chlorophyll during most part of the year, except in May, when highest DIN was recorded in the southern part. Statistical analysis revealed that dissolved NH 4 + –N and urea could combinedly explain 43% of Chlorophyll- a (Chl- a ) variability which was relatively higher than that explained by NO 3 − –N and NO 2 − –N (12.4%) in lagoon water. Trophic state index calculated for different sectors of the lagoon confirmed the inter-sectoral and inter-seasonal shift from mesotrophic to eutrophic conditions largely depending on nutrient rich freshwater input.
Anticancer Properties of Highly Purified l-Asparaginase from Withania somnifera L. against Acute Lymphoblastic Leukemia
Withania somnifera L. has been traditionally used as a sedative and hypnotic. The present study was carried out for the purification, characterization, and in vitro cytotoxicity of l-asparaginase from W. somnifera L. l-Asparaginase was purified from the fruits of W. somnifera L. up to 95% through chromatography. The purified l-asparaginase was characterized by size exclusion chromatography, polyacrylamide gel electrophoresis (PAGE), and 2D PAGE. The antitumor and growth inhibition effect of the l-asparaginase was assessed using [3-(4, 5-dimethyl-thiazol-2yl)-2, 5-diphenyl-tetrazolium bromide] (MTT) colorimetric dye reduction method. The purified enzyme is a homodimer, with a molecular mass of 72 ± 0.5 kDa, and the pI value of the enzyme was around 5.1. This is the first report of the plant containing l-asparaginase with antitumor activity. Data obtained from the MTT assay showed a LD₅₀ value of 1.45 ± 0.05 IU/ml. W. somnifera L. proved to be an effective and a novel source of l-asparaginase. Furthermore, it shows a lot of similarity with bacterial l-asparaginases EC-2.
Characterization of a Recombinant Glutaminase-Free L-Asparaginase (ansA3) Enzyme with High Catalytic Activity from Bacillus licheniformis
L-Asparaginase (3.5.1.1) is an enzyme widely used to treat the acute lymphoblastic leukemia. Two genes coding for L-asparaginase (ansA1 and ansA3) from Bacillus licheniformis MTCC 429 were cloned and overexpressed in Escherichia coli BL21 (DE3) cells. The recombinant proteins were purified to homogeneity by one-step purification process and further characterized for various biochemical parameters. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed that both the enzymes are monomers of ∼37 kDa. Recombinant ansA1 was found to be highly unstable, and recombinant ansA3 was catalytically active and stable, which showed an optimum activity of 407.65 IU/mg at 37 °C and pH 8. Recombinant ansA3 showed higher substrate specificity for L-asparagine with negligible glutaminase activity. Kinetic parameters like K ₘ , Vₘₐₓ, kcₐₜ, and kcₐₜ/K ₘ were calculated for recombinant ansA3.
Influence of suspended particulate matter on nutrient biogeochemistry of a tropical shallow lagoon, Chilika, India
The behaviour of suspended particulate matter (SPM), salinity profile, dissolved nutrients, total (T.Chl- a ) and size fractionated chlorophyll- a (F.Chl- a ) were studied seasonally at Chilika Lagoon, east coast of India, during 2008–2009. The study showed large spatio-temporal variations among these parameters. The concentration of dissolved inorganic nitrogen and inorganic phosphate were found to be maximum during the monsoon, followed by post- and pre-monsoon, although the mean N:P ratios, which indicate the relative availability of N with respect to P, were 9.13 ± 3.09, 16.57 ± 11.53 and 5.47 ± 3.13, respectively. It was evident from the results that during pre-monsoon and postmonsoon, the lagoon exhibits nitrogen limitation. Mean T.Chl- a biomass in the lagoon showed distinct seasonality with maximum values during the pre-monsoon (23.12 ± 9.75 mg m −3 ) followed by monsoon and post-monsoon. Irrespective of seasons, maximum T.Chl- a was found in the northern part of the lagoon. SPM concentrations during the monsoon were relatively higher in the freshwater dominated zones compared to seawater dominated areas, indicating its riverine sources. The correlation between SPM and various dissolved nutrients ( p  < 0.05) suggests its influence on the physico-chemical conditions at varying levels. It is summarized that seasonal variation of SPM and nutrients contributed by rivers, wind induced re-suspension events and in situ  regeneration processes play a crucial role in the lagoon biogeochemical cycle.
Storm surge vulnerability along Chennai–Cuddalore coast due to a severe cyclone THANE
A very severe cyclonic storm ‘Thane’ developed over the Bay of Bengal during 25–30 December 2011, crossed the Tamilnadu coast between Pondicherry and Cuddalore (southeast coast of India) in early hours of 30 December with a wind speed 120–140 km/h. The offshore tide record reveals that the surge started to generate around 1100 hours on 29 December 2011 with a height 0.4 m and later raised to 0.68 m at the time of land fall, that is, early hours (0000 hours) of 30 December 2011. Field reconnaissance survey on surge run-up and inundation distance at 15 selected locations of cyclone affected areas reveals that the vulnerability levels are highly variable along the coast. The inundation distance extended up to 30–230 m landward from the shoreline and run-up reached to 1.6–3.2 m above chart datum depending upon the cross-shore geometry of the location. In the areas (Verranampattinam, Chinamudaliyar kuppam and Silver beach) near cyclone landfall, the run-up was up to 2.5–3.2 m and the inundation distance extended up to a maximum of 230 m. However, in the areas located about 150 km north of cyclone landfall, the run-up limited from 1.6 to 1.7 m and the inundation distance extended up to only a maximum of 169 m. The inundation distance is mainly influenced by the slope of the beach. In the areas having beach slope between 1 in 135 and 1 in 220, the inundation distance was 210–230 m. However, in the areas having beach slope 1 in 17 to 1 in 34, the inundation distance is restricted between 35 and 50 m.