Papers by Shanmugasundram Senthil Balan
Process Biochemistry, 2016
Achievements in the Life Sciences, 2016
The use and commercial applications of biosurfactants in the petroleum industries have been raise... more The use and commercial applications of biosurfactants in the petroleum industries have been raised during the past decades. Marine bacteria and their efficiency in crude oil recovery has been less studied than terrestrial strain, hence this present study. A novel marine bacterium Bacillus simplex having promising biosurfactant production was isolated from a petroleum hydrocarbon-contaminated coastal sea sediment samples of Nagapattinam fishing harbor, Tamil Nadu, India. This strain showed most economical biosurfactant production with an agro-industrial waste substrate, sunflower oil cake at 54th h time incubation along with the cultural conditions of 20 ppt salinity, 35°C temperature, and pH 7. The produced biosurfactant was purified, which was accounted at 908 ± 7 mg/L on dry weight basis. The biosurfactant was identified as lipopeptide with a molecular mass of 1111.1 Da which was deduced using TLC, biochemical estimation methods, FT-IR, NMR, and MALDI-TOF MS analysis. Furthermore, this purified lipopeptide surfactant showed consistent and enhanced crude oil recovering efficiency under different salinity conditions (0-30%). Based on the above facts, the isolated novel marine bacterium proved its cheaper production of novel biosurfactant and its promising oil recovering efficiency even at hypersaline conditions. Further, this is the first report of a biosurfactant from the bacterium Bacillus simplex.
Asian Pacific Journal of Tropical Biomedicine, 2012
Abstract Objective In this study, gelatinase producing bacteria were probed from sediment samples... more Abstract Objective In this study, gelatinase producing bacteria were probed from sediment samples of Porto Novo Coastal sites, India. Screening and identification of potential strain were done followed by optimization of physico-chemical parameters; bulk production and gelatinase extraction were carried out. Methods For probing of gelatinase potential producer primary and secondary screening was carried out for qualitative and quantitative estimation. Optimization of physico-chemical parameters for improved production of gelatinase enzyme and large scale of gelatinase was produced. Gelatinase precipitation was standardized using different saturation rates of ammonium sulphate from 10 to 100% at 4°C. Results There were 8 morphologically different gelatinase producing bacteria were initially delved through primary screening tests. Bacillus spp produced maximum gelatinase activity (2.1U/mL) in secondary screening test. Optimizing its abiotic and biotic factors, maximum enzyme activity was achieved at 48h incubation period (2.2U/mL), 2.5 pH (2.5U/mL), 35°C temperature (2.55U/mL), 0.8% lactose (2.6U/mL), 1.4% gelatin (2.9U/mL) as the ideal carbon source and nitrogen source, 1% salinity (2.9U/mL) and 3ml of inoculum containing 5.6×106/mL (3.3U/mL). From the optimized factors, bulk production was carried out and saturation rate of 40% ammonium sulphate, precipitated out maximum enzyme with lowered dry weight indicates its enzyme purity and recovered enzyme showed 4.1U/mg of activity. Conclusion The study revealed that the isolated strain Bacillus spp has its potentiality for industrial scale production and the results will stand as a base line data for the application of gelatinase in future.
Journal of Microbiology and Biotechnology, 2013
The present study dealt with the isolation, identification and enzyme characterization of potenti... more The present study dealt with the isolation, identification and enzyme characterization of potential luminous bacteria from water, sediment, squid, and cuttle fish samples of the Karaikal coast, Bay of Bengal, India during the study period September 2007-August 2008. Bioluminescent strains were screened in SWC agar and identified using biochemical tests. As Shewanella henadai was found to be the most common and abundant species with maximum light emission [69,702,240 photons per second (pps)], the optimum ranges of various physicochemical parameters that enhance the luciferase activity in Shewanella hanedai were worked out. The maximum luciferase activity was observed at the temperature of 25 o C (69,674,387 pps), pH of 8.0 (70,523,671 pps), salinity of 20 ppt (71,674,387 pps), incubation period of 16 h (69,895,714 pps), 4% peptone (70,895,152 pps) as nitrogen source, 0.9% glycerol (71,625,196 pps), and the ionic supplements of 0.3% CaCO 3 (73,991,591 pps), 0.3% K 2 HPO 4 (73,919,915 pps), and 0.2% MgSO 4 (72,161,155 pps). Shewanella hanedai was cultured at optimum ranges for luciferase enzyme characterization. From the centrifuged supernatant, the proteins were precipitated with 60% ammonium sulfate, dialyzed, and purified using anionexchange chromatography, and then luciferase was eluted with 500 mM phosphate of pH 7.0. The purified luciferase enzyme was subjected to SDS-PAGE and the molecular mass was determined as 78 kDa.
African Journal of Microbiology Research, 2012
The rapidly increasing importance of enzyme urease applications has drawn attention to the need o... more The rapidly increasing importance of enzyme urease applications has drawn attention to the need of enzyme study. It prompted the present study to hunt a promising bacterial strain with the desired nature from the Porto Novo coast. Urease enzyme is used in diagnostic kits for measuring blood urea, removal of urea from alcoholic beverages, urease conductometric biosensors for detection of heavy metal ions, etc. In our study, urease enzyme produced by marine bacteria, was isolated, identified and characterized. Samples of water and sediment from Porto Novo coast were taken for analysis. After that, sediment sample showed more number of urease producing bacteria when compared to water sample. Urease producing bacteria were identified as Klebseilla spp, Proteus spp, Lactobacillus spp and Streptococcus spp, from that urease producer Klebseilla spp showed maximum urease production through phenol hypochlorite assay. The potential strain was optimized with their physiochemical parameters showing maximum urease production at 48 h incubation, pH 7, temperature 35°C, salinity 20 ppt producing 1.7, 1.72, 1.67 and 1.72 U/ml, respectively. An optimization with different carbon and nitrogen source showed maximum at 0.7% glucose with 2.05 U/ml and 0.7% peptone with 2.15 U/ml. Mineral supplements 0.03% sodium acetate, 0.04% potassium dihydrogen phosphate, 0.04% nickel sulphate and 0.04% magnesium sulphate showed maximum production of 2.18, 2.27, 2.37 and 2.4 U/ml, respectively. Urea, the enzyme substrate showed maximum production at 0.3% with 2.25 U/ml urease production. Final optimization with inoculum size showed maximum production with increased volume of inoculum with 50 ml of 6-8 × 10 7 cell/ml to 500 ml of fermentation medium, showed 2.65 U/ml of urease on 36 h of incubation. Urease enzyme was produced in fermentation medium with the above optimized condition and showed a production of 2.8 U/ml. Then the enzyme was partially purified using dialysis membrane after being fractionated with 60% ammonium sulphate at 5°C. The characterization of urease enzyme from marine bacterium Klebsiella species and the optimization of various physicochemical factors for maximum urease production and its activity stand as a ready reference for more elaborate work of this line in future. The results of the present study will be a base line data for the application of this urease.
3 Biotech, 2016
Biosurfactants have gained a renewed interest in the recent years for their commercial applicatio... more Biosurfactants have gained a renewed interest in the recent years for their commercial application in diverse research areas. Recent evidences suggest that the antimicrobial activities exhibited by biosurfactants make them promising molecules for the application in the field of therapeutics. Marine microbes are well known for their unique metabolic and functional properties; however, few reports are available till date regarding their biosurfactant production and antimicrobial potential. In an ongoing survey for bioactive microbial metabolites from microbes isolated from diverse ecological niches, a marine Staphylococcus saprophyticus SBPS 15 isolated from the petroleum hydrocarbon contaminated coastal site, Puducherry, India, was identified as a promising biosurfactant producer based on multiple screening methods. This bacterium exhibited growth-dependent biosurfactant production and the recorded yield was 1.345 ± 0.056 g/L (on dry weight basis). The biosurfactant was purified and chemically characterized as a glycolipid with a molecular mass of 606.7 Da, based on TLC, biochemical estimation methods, FT-IR spectrum and MALDI-TOF-MS analysis. Further, the estimated molecular mass was different from the earlier reports on biosurfactants. This new glycolipid biosurfactant exhibited a board range of pH and temperature stability. Furthermore, it revealed a promising antimicrobial activity against many tested human pathogenic bacterial and fungal clinical isolates. Based on these observations, the isolated biosurfactant from the marine S. saprophyticus revealed board physicochemical stabilities and possess excellent antimicrobial activities which proves its significance for possible use in various therapeutic and biomedical applications. To the best of our knowledge, this is the first report of a biosurfactant from the bacterium, S. saprophyticus.
In an ongoing survey for bioactive microbial metabolites from different biospheres of India, a ne... more In an ongoing survey for bioactive microbial metabolites from different biospheres of India, a new marine bacterium identified as Pontibacter korlensis strain SBK-47 was isolated from the coastal waters of Karaikal, Puducherry, India, which produced a novel lipopeptide biosurfactant. The biosurfactant was purified and structurally elucidated as Palmitic acid-Ser-Asp-Val-Ser-Ser based on TLC, FT-IR, NMR, GC–MS, HPLC, MALDI-TOF and tandem MS analysis. This novel lipopeptide biosurfactant was named as Pontifactin. Pontifactin exhibited a surface tension reduction and critical micelle concentration (CMC) of 25 mN/m and 25 mg/L, respectively. Furthermore, the biosurfactant showed emulsifi-cation and surface tension stability over a wide range of pH (4–10) and temperature up to 100 °C. Pontifactin showed promising antimicrobial activity against Streptococcus mutans, Micrococcus luteus, Salmonella typhi and Klebsiella oxy-toca at a biosurfactant concentration ranging between 1 and 2 mg/mL and maximum anti-biofilm activity at the biosur-factant concentration of 2 mg/mL against Bacillus subtilis, Staphylococcus aureus, Salmonella typhi and Vibrio cholerae. This is the first report on Pontifactin, a multifunctional lipopeptide biosurfactant, produced by a marine Pontibacter kor-lensis strain SBK-47, exhibiting promising surface-active, antimicrobial and anti-biofilm activities and thus finds possible use in biomedical applications.
The present study dealt with the isolation, identification
and enzyme characterization of potenti... more The present study dealt with the isolation, identification
and enzyme characterization of potential luminous bacteria
from water, sediment, squid, and cuttle fish samples of the
Karaikal coast, Bay of Bengal, India during the study
period September 2007 – August 2008. Bioluminescent
strains were screened in SWC agar and identified using
biochemical tests. As Shewanella henadai was found to be
the most common and abundant species with maximum
light emission [69,702,240 photons per second (pps)], the
optimum ranges of various physicochemical parameters that
enhance the luciferase activity in Shewanella hanedai were
worked out. The maximum luciferase activity was observed
at the temperature of 25oC (69,674,387 pps), pH of 8.0
(70,523,671 pps), salinity of 20 ppt (71,674,387 pps), incubation
period of 16 h (69,895,714 pps), 4% peptone (70,895,152 pps)
as nitrogen source, 0.9% glycerol (71,625,196 pps), and the
ionic supplements of 0.3% CaCO3 (73,991,591 pps), 0.3%
K2HPO4 (73,919,915 pps), and 0.2% MgSO4 (72,161,155 pps).
Shewanella hanedai was cultured at optimum ranges for
luciferase enzyme characterization. From the centrifuged
supernatant, the proteins were precipitated with 60%
ammonium sulfate, dialyzed, and purified using anion-
exchange chromatography, and then luciferase was eluted
with 500 mM phosphate of pH 7.0. The purified luciferase
enzyme was subjected to SDS-PAGE and the molecular
mass was determined as 78 kDa.
Objective: In this study, gelatinase producing bacteria were probed from sediment samples of Po... more Objective: In this study, gelatinase producing bacteria were probed from sediment samples of Porto Novo Coastal sites, India. Screening and identification of potential strain were done followed by optimization of physico-chemical parameters; bulk production and gelatinase extraction
were carried out.
Methods: For probing of gelatinase potential producer primary and secondary screening was carried out for qualitative and quantitative estimation. Optimization of physico- chemical parameters for improved production of gelatinase enzyme and large scale of gelatinase was produced. Gelatinase precipitation was standardized using different saturation rates of ammonium sulphate from 10 to 100% at 4.Results: There were 8 morphological different gelatinase producing bacteria were initially delved through primary screening tests. Bacillus
sp. produced maximum gelatinase activity (2.1U/mL) in secondary screening test. Optimizing its abiotic and biotic factors, maximum enzyme activity was achieved at 48h incubation period
(2.2U/mL), 2.5 pH (2.5U/mL), 35 degree Celsius temperature (2.55U/mL), 0.8% lactose (2.6U/mL), 1.4% gelatin
(2.9U/mL) as the ideal carbon source and nitrogen source, 1% salinity (2.9U/mL) and 3ml of inoculum containing 5.6 multiplied by 106/ mL (3.3U/mL). From the optimized factors, bulk production was
carried out and saturation rate of 40% ammonium sulphate, precipitated out maximum enzyme with lowered dry weight indicates its enzyme purity and recovered enzyme showed 4.1U/mg of activity. Conclusion: The study revealed that the isolated strain Bacillus spp has its otentiality
for industrial scale production and the results will stand as a base line data for the application of gelatinase in future.
The rapidly increasing importance of enzyme urease applications has drawn attention to the need o... more The rapidly increasing importance of enzyme urease applications has drawn attention to the need of enzyme study. It prompted the present study to hunt a promising bacterial strain with the desired nature from the Porto Novo coast. Urease enzyme is used in diagnostic kits for measuring blood urea, removal of urea from alcoholic beverages, urease conductometric biosensors for detection of heavy metal ions, etc. In our study, urease enzyme produced by marine bacteria, was isolated, identified and characterized. Samples of water and sediment from Porto Novo coast were taken for analysis. After that, sediment sample showed more number of urease producing bacteria when compared to water sample. Urease producing bacteria were identified as Klebseilla spp, Proteus spp, Lactobacillus spp and Streptococcus spp, from that urease producer Klebseilla spp showed maximum urease production through phenol hypochlorite assay. The potential strain was optimized with their physiochemical parameters showing maximum urease production at 48 h incubation, pH 7, temperature 35°C, salinity 20 ppt producing 1.7, 1.72, 1.67 and 1.72 U/ml, respectively. An optimization with different carbon and nitrogen source showed maximum at 0.7% glucose with 2.05 U/ml and 0.7% peptone with 2.15 U/ml. Mineral supplements 0.03% sodium acetate, 0.04% potassium dihydrogen phosphate, 0.04% nickel sulphate and 0.04% magnesium sulphate showed maximum production of 2.18, 2.27, 2.37 and 2.4 U/ml, respectively. Urea, the enzyme substrate showed maximum production at 0.3% with 2.25 U/ml urease production. Final optimization with inoculum size showed maximum production with increased volume of inoculum with 50 ml of 6-8 × 10 7 cell/ml to 500 ml of fermentation medium, showed 2.65 U/ml of urease on 36 h of incubation. Urease enzyme was produced in fermentation medium with the above optimized condition and showed a production of 2.8 U/ml. Then the enzyme was partially purified using dialysis membrane after being fractionated with 60% ammonium sulphate at 5°C. The characterization of urease enzyme from marine bacterium Klebsiella species and the optimization of various physico-chemical factors for maximum urease production and its activity stand as a ready reference for more elaborate work of this line in future. The results of the present study will be a base line data for the application of this urease.
Biosurfactants have gained a renewed interest in the recent years for their commercial applicatio... more Biosurfactants have gained a renewed interest in the recent years for their commercial application in diverse research areas. Recent evidences suggest that the antimicrobial activities exhibited by biosurfactants make them promising molecules for the application in the field of therapeutics. Marine microbes are well known for their unique metabolic and functional properties; however, few reports are available till date regarding their biosurfactant production and antimicrobial potential. In an ongoing survey for bioactive microbial metabolites from microbes isolated from diverse ecological niches, a marine Staphylococcus saprophyticus SBPS 15 isolated from the petroleum hydrocarbon contaminated coastal site, Pudu-cherry, India, was identified as a promising biosurfactant producer based on multiple screening methods. This bacterium exhibited growth-dependent biosurfactant production and the recorded yield was 1.345 ± 0.056 g/L (on dry weight basis). The biosurfactant was purified and chemically characterized as a glycolipid with a molecular mass of 606.7 Da, based on TLC, biochemical estimation methods, FT-IR spectrum and MALDI-TOF–MS analysis. Further, the estimated molecular mass was different from the earlier reports on biosurfactants. This new glycolipid biosurfactant exhibited a board range of pH and temperature stability. Furthermore, it revealed a promising antimicrobial activity against many tested human patho-genic bacterial and fungal clinical isolates. Based on these observations, the isolated biosurfactant from the marine S. saprophyticus revealed board physicochemical stabilities and possess excellent antimicrobial activities which proves its significance for possible use in various therapeutic and biomedical applications. To the best of our knowledge, this is the first report of a biosurfactant from the bacterium, S. saprophyticus.
The use and commercial applications of biosurfactants in the petroleum industries have been raise... more The use and commercial applications of biosurfactants in the petroleum industries have been raised during the past decades. Marine bacteria and their efficiency in crude oil recovery has been less studied than terrestrial strain, hence this present study. A novel marine bacterium Bacillus simplex having promising biosurfactant production was isolated from a petroleum hydrocarbon-contaminated coastal sea sediment samples of Nagapattinam fishing harbor, Tamil Nadu, India. This strain showed most economical biosurfactant production with an agro-industrial waste substrate, sunflower oil cake at 54th h time incubation along with the cultural conditions of 20 ppt salinity, 35 °C temperature, and pH 7. The produced biosurfactant was purified, which was accounted at 908 ± 7 mg/L on dry weight basis. The biosurfactant was identified as lipopeptide with a molecular mass of 1111.1 Da which was deduced using TLC, biochemical estimation methods, FT-IR, NMR, and MALDI-TOF MS analysis. Furthermore, this purified lipopeptide surfactant showed consistent and enhanced crude oil recovering efficiency under different salinity conditions (0–30%). Based on the above facts, the isolated novel marine bacterium proved its cheaper production of novel biosurfactant and its promising oil recovering efficiency even at hypersaline conditions. Further, this is the first report of a biosurfactant from the bacterium Bacillus simplex.
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Papers by Shanmugasundram Senthil Balan
and enzyme characterization of potential luminous bacteria
from water, sediment, squid, and cuttle fish samples of the
Karaikal coast, Bay of Bengal, India during the study
period September 2007 – August 2008. Bioluminescent
strains were screened in SWC agar and identified using
biochemical tests. As Shewanella henadai was found to be
the most common and abundant species with maximum
light emission [69,702,240 photons per second (pps)], the
optimum ranges of various physicochemical parameters that
enhance the luciferase activity in Shewanella hanedai were
worked out. The maximum luciferase activity was observed
at the temperature of 25oC (69,674,387 pps), pH of 8.0
(70,523,671 pps), salinity of 20 ppt (71,674,387 pps), incubation
period of 16 h (69,895,714 pps), 4% peptone (70,895,152 pps)
as nitrogen source, 0.9% glycerol (71,625,196 pps), and the
ionic supplements of 0.3% CaCO3 (73,991,591 pps), 0.3%
K2HPO4 (73,919,915 pps), and 0.2% MgSO4 (72,161,155 pps).
Shewanella hanedai was cultured at optimum ranges for
luciferase enzyme characterization. From the centrifuged
supernatant, the proteins were precipitated with 60%
ammonium sulfate, dialyzed, and purified using anion-
exchange chromatography, and then luciferase was eluted
with 500 mM phosphate of pH 7.0. The purified luciferase
enzyme was subjected to SDS-PAGE and the molecular
mass was determined as 78 kDa.
were carried out.
Methods: For probing of gelatinase potential producer primary and secondary screening was carried out for qualitative and quantitative estimation. Optimization of physico- chemical parameters for improved production of gelatinase enzyme and large scale of gelatinase was produced. Gelatinase precipitation was standardized using different saturation rates of ammonium sulphate from 10 to 100% at 4.Results: There were 8 morphological different gelatinase producing bacteria were initially delved through primary screening tests. Bacillus
sp. produced maximum gelatinase activity (2.1U/mL) in secondary screening test. Optimizing its abiotic and biotic factors, maximum enzyme activity was achieved at 48h incubation period
(2.2U/mL), 2.5 pH (2.5U/mL), 35 degree Celsius temperature (2.55U/mL), 0.8% lactose (2.6U/mL), 1.4% gelatin
(2.9U/mL) as the ideal carbon source and nitrogen source, 1% salinity (2.9U/mL) and 3ml of inoculum containing 5.6 multiplied by 106/ mL (3.3U/mL). From the optimized factors, bulk production was
carried out and saturation rate of 40% ammonium sulphate, precipitated out maximum enzyme with lowered dry weight indicates its enzyme purity and recovered enzyme showed 4.1U/mg of activity. Conclusion: The study revealed that the isolated strain Bacillus spp has its otentiality
for industrial scale production and the results will stand as a base line data for the application of gelatinase in future.
and enzyme characterization of potential luminous bacteria
from water, sediment, squid, and cuttle fish samples of the
Karaikal coast, Bay of Bengal, India during the study
period September 2007 – August 2008. Bioluminescent
strains were screened in SWC agar and identified using
biochemical tests. As Shewanella henadai was found to be
the most common and abundant species with maximum
light emission [69,702,240 photons per second (pps)], the
optimum ranges of various physicochemical parameters that
enhance the luciferase activity in Shewanella hanedai were
worked out. The maximum luciferase activity was observed
at the temperature of 25oC (69,674,387 pps), pH of 8.0
(70,523,671 pps), salinity of 20 ppt (71,674,387 pps), incubation
period of 16 h (69,895,714 pps), 4% peptone (70,895,152 pps)
as nitrogen source, 0.9% glycerol (71,625,196 pps), and the
ionic supplements of 0.3% CaCO3 (73,991,591 pps), 0.3%
K2HPO4 (73,919,915 pps), and 0.2% MgSO4 (72,161,155 pps).
Shewanella hanedai was cultured at optimum ranges for
luciferase enzyme characterization. From the centrifuged
supernatant, the proteins were precipitated with 60%
ammonium sulfate, dialyzed, and purified using anion-
exchange chromatography, and then luciferase was eluted
with 500 mM phosphate of pH 7.0. The purified luciferase
enzyme was subjected to SDS-PAGE and the molecular
mass was determined as 78 kDa.
were carried out.
Methods: For probing of gelatinase potential producer primary and secondary screening was carried out for qualitative and quantitative estimation. Optimization of physico- chemical parameters for improved production of gelatinase enzyme and large scale of gelatinase was produced. Gelatinase precipitation was standardized using different saturation rates of ammonium sulphate from 10 to 100% at 4.Results: There were 8 morphological different gelatinase producing bacteria were initially delved through primary screening tests. Bacillus
sp. produced maximum gelatinase activity (2.1U/mL) in secondary screening test. Optimizing its abiotic and biotic factors, maximum enzyme activity was achieved at 48h incubation period
(2.2U/mL), 2.5 pH (2.5U/mL), 35 degree Celsius temperature (2.55U/mL), 0.8% lactose (2.6U/mL), 1.4% gelatin
(2.9U/mL) as the ideal carbon source and nitrogen source, 1% salinity (2.9U/mL) and 3ml of inoculum containing 5.6 multiplied by 106/ mL (3.3U/mL). From the optimized factors, bulk production was
carried out and saturation rate of 40% ammonium sulphate, precipitated out maximum enzyme with lowered dry weight indicates its enzyme purity and recovered enzyme showed 4.1U/mg of activity. Conclusion: The study revealed that the isolated strain Bacillus spp has its otentiality
for industrial scale production and the results will stand as a base line data for the application of gelatinase in future.