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Enhanced production and characterization of a solvent stable amylase from solvent tolerant Bacillus tequilensis RG-01: thermostable and surfactant resistant.

Tiwari S, Shukla N, Mishra P, Gaur R - ScientificWorldJournal (2014)

Bottom Line: The enzyme was showed it 100% activity at 55°C and pH 7.0 with 119% and 127% stability at 55°C and pH 7.0, respectively.The enzyme was also stable in the presence of SDS, Tween-40, Tween-60, and Tween-80 (1%) and was found stimulatory effect, respectively.Only Triton-X-100 showed a moderate inhibitory effect (5%) on amylase activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Centre of Excellence, Dr. Ram Manohar Lohia Avadh University, Faizabad, Uttar Pradesh 224001, India.

ABSTRACT
Ten bacterial strains isolated from the soil samples in the presence of cyclohexane were screened for amylase production. Among them, culture RG-01 was adjudged as the best amylase producer and was identified as Bacillus tequilensis from MTCC, Chandigarh. The isolate showed maximum amylase production (8100 U/mL) in the presence of starch, peptone, and Ca(2+) ions at 55°C pH 7.0 within 24 h of incubation. The enzyme was stable in the presence of n-dodecane, isooctane, n-decane, xylene, toluene, n-hexane, n-butanol, and cyclohexane, respectively. The presence of benzene, methanol, and ethanol marginally reduced the amylase stability, respectively. The enzyme was showed it 100% activity at 55°C and pH 7.0 with 119% and 127% stability at 55°C and pH 7.0, respectively. The enzyme was also stable in the presence of SDS, Tween-40, Tween-60, and Tween-80 (1%) and was found stimulatory effect, respectively. Only Triton-X-100 showed a moderate inhibitory effect (5%) on amylase activity. This isolate (Bacillus tequilensis RG-01) may be useful in several industrial applications owing to its thermotolerant and organic solvents and surfactants resistance characteristics.

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Related in: MedlinePlus

Effect of incubation periods on amylase production and stability. The flasks were inoculated with culture and were incubated at different incubation periods (4–48 h) at initial pH 7.0, 55°C. For enzyme activity, the reaction was assayed at respective incubation periods and for stability enzyme was preincubated for 4–48 h at 55°C for 1 h and assayed by standard assay method. Error bars presented mean values of ±standard deviation of triplicates of three independent experiments.
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fig5: Effect of incubation periods on amylase production and stability. The flasks were inoculated with culture and were incubated at different incubation periods (4–48 h) at initial pH 7.0, 55°C. For enzyme activity, the reaction was assayed at respective incubation periods and for stability enzyme was preincubated for 4–48 h at 55°C for 1 h and assayed by standard assay method. Error bars presented mean values of ±standard deviation of triplicates of three independent experiments.

Mentions: Just after optimization of temperature for amylase production, in the liquid medium, incubation period was optimized for enzyme production and stability. The results clearly indicated that Bacillus tequilensis RG-01 showed 4800 U/mL enzyme activity with 125% stability in 20 h of incubation (Figure 5). Further increase in the incubation period did not increase the enzyme production, but the stability of enzyme is 100% in 72 h (data not shown). In contrast to our results, Özdemir et al. [23] reported that α-amylase production by B. subtilis was maximum (2902 U/mg) in 72 h, after which a gradual decrease was observed. It may be due to denaturation or decomposition of α-amylase owing to interaction with other components in the medium, as it is reported elsewhere [24]. Incubation time depends on the characteristics of the culture, on growth rate, and enzyme production [25]. Moreover, the reaction for maximum enzyme production at 72 h could be due to the fact that the microorganism was in its exponential phase. At later stage, when nutrients are depleted, it reaches its stationary phase and can start to produce secondary metabolites, thus resulting in a lower yield of enzyme [26]. Thus, our strain producing α-amylase within 20 h of incubation is better than that reported by the other workers mentioned above.


Enhanced production and characterization of a solvent stable amylase from solvent tolerant Bacillus tequilensis RG-01: thermostable and surfactant resistant.

Tiwari S, Shukla N, Mishra P, Gaur R - ScientificWorldJournal (2014)

Effect of incubation periods on amylase production and stability. The flasks were inoculated with culture and were incubated at different incubation periods (4–48 h) at initial pH 7.0, 55°C. For enzyme activity, the reaction was assayed at respective incubation periods and for stability enzyme was preincubated for 4–48 h at 55°C for 1 h and assayed by standard assay method. Error bars presented mean values of ±standard deviation of triplicates of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4226188&req=5

fig5: Effect of incubation periods on amylase production and stability. The flasks were inoculated with culture and were incubated at different incubation periods (4–48 h) at initial pH 7.0, 55°C. For enzyme activity, the reaction was assayed at respective incubation periods and for stability enzyme was preincubated for 4–48 h at 55°C for 1 h and assayed by standard assay method. Error bars presented mean values of ±standard deviation of triplicates of three independent experiments.
Mentions: Just after optimization of temperature for amylase production, in the liquid medium, incubation period was optimized for enzyme production and stability. The results clearly indicated that Bacillus tequilensis RG-01 showed 4800 U/mL enzyme activity with 125% stability in 20 h of incubation (Figure 5). Further increase in the incubation period did not increase the enzyme production, but the stability of enzyme is 100% in 72 h (data not shown). In contrast to our results, Özdemir et al. [23] reported that α-amylase production by B. subtilis was maximum (2902 U/mg) in 72 h, after which a gradual decrease was observed. It may be due to denaturation or decomposition of α-amylase owing to interaction with other components in the medium, as it is reported elsewhere [24]. Incubation time depends on the characteristics of the culture, on growth rate, and enzyme production [25]. Moreover, the reaction for maximum enzyme production at 72 h could be due to the fact that the microorganism was in its exponential phase. At later stage, when nutrients are depleted, it reaches its stationary phase and can start to produce secondary metabolites, thus resulting in a lower yield of enzyme [26]. Thus, our strain producing α-amylase within 20 h of incubation is better than that reported by the other workers mentioned above.

Bottom Line: The enzyme was showed it 100% activity at 55°C and pH 7.0 with 119% and 127% stability at 55°C and pH 7.0, respectively.The enzyme was also stable in the presence of SDS, Tween-40, Tween-60, and Tween-80 (1%) and was found stimulatory effect, respectively.Only Triton-X-100 showed a moderate inhibitory effect (5%) on amylase activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Centre of Excellence, Dr. Ram Manohar Lohia Avadh University, Faizabad, Uttar Pradesh 224001, India.

ABSTRACT
Ten bacterial strains isolated from the soil samples in the presence of cyclohexane were screened for amylase production. Among them, culture RG-01 was adjudged as the best amylase producer and was identified as Bacillus tequilensis from MTCC, Chandigarh. The isolate showed maximum amylase production (8100 U/mL) in the presence of starch, peptone, and Ca(2+) ions at 55°C pH 7.0 within 24 h of incubation. The enzyme was stable in the presence of n-dodecane, isooctane, n-decane, xylene, toluene, n-hexane, n-butanol, and cyclohexane, respectively. The presence of benzene, methanol, and ethanol marginally reduced the amylase stability, respectively. The enzyme was showed it 100% activity at 55°C and pH 7.0 with 119% and 127% stability at 55°C and pH 7.0, respectively. The enzyme was also stable in the presence of SDS, Tween-40, Tween-60, and Tween-80 (1%) and was found stimulatory effect, respectively. Only Triton-X-100 showed a moderate inhibitory effect (5%) on amylase activity. This isolate (Bacillus tequilensis RG-01) may be useful in several industrial applications owing to its thermotolerant and organic solvents and surfactants resistance characteristics.

Show MeSH
Related in: MedlinePlus