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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 pH on amylase production, activity, and stability. The flasks were inoculated with culture and were incubated at different pH (5–12) for 24 h at 55°C. For enzyme activity, the reaction was assayed at respective pH and for stability enzyme was preincubated with buffers (100 mM, in ratio 1 : 1) of different pH (5–12) 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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fig4: Effect of pH on amylase production, activity, and stability. The flasks were inoculated with culture and were incubated at different pH (5–12) for 24 h at 55°C. For enzyme activity, the reaction was assayed at respective pH and for stability enzyme was preincubated with buffers (100 mM, in ratio 1 : 1) of different pH (5–12) 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: Among the physical parameters, the pH of the growth medium plays an important role by inducing morphological change in the organism and in enzyme secretion [19, 20]. Different pH, viz, 5.0 to 12.0 in the medium, was tested for amylase production, activity, and stability by Bacillus tequilensis RG-01 at their optimal temperature and incubation period. Bacillus tequilensis RG-01 showed 5200 U/mL enzyme productions, 100% activity, and 127% stability at pH 7.0 (Figure 4). Most of the Bacillus strains used commercially for the production of bacterial α-amylases have an optimum pH between 6.0 and 7.0 for growth and enzyme production [4, 18]. Strain RG-01 showed 86% activity and 91% stability at pH 9.0 (Figure 4). Haq et al. [21] reported pH 7.5–8.0 to be the best for the production of alpha amylase by Bacillus subtilis. Further increase and decrease in the medium pH reduced the enzyme production, activity, and stability (Figure 4). The pH change observed during the growth of the organism also affects product stability in the medium. Results show that enzyme production was generally active and stable from pH 5.0 to 10.0, which indicates excellent buffering property. The pH values also serve as a valuable indicator of the initiation and end of enzyme synthesis [22].


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 pH on amylase production, activity, and stability. The flasks were inoculated with culture and were incubated at different pH (5–12) for 24 h at 55°C. For enzyme activity, the reaction was assayed at respective pH and for stability enzyme was preincubated with buffers (100 mM, in ratio 1 : 1) of different pH (5–12) 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

fig4: Effect of pH on amylase production, activity, and stability. The flasks were inoculated with culture and were incubated at different pH (5–12) for 24 h at 55°C. For enzyme activity, the reaction was assayed at respective pH and for stability enzyme was preincubated with buffers (100 mM, in ratio 1 : 1) of different pH (5–12) 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: Among the physical parameters, the pH of the growth medium plays an important role by inducing morphological change in the organism and in enzyme secretion [19, 20]. Different pH, viz, 5.0 to 12.0 in the medium, was tested for amylase production, activity, and stability by Bacillus tequilensis RG-01 at their optimal temperature and incubation period. Bacillus tequilensis RG-01 showed 5200 U/mL enzyme productions, 100% activity, and 127% stability at pH 7.0 (Figure 4). Most of the Bacillus strains used commercially for the production of bacterial α-amylases have an optimum pH between 6.0 and 7.0 for growth and enzyme production [4, 18]. Strain RG-01 showed 86% activity and 91% stability at pH 9.0 (Figure 4). Haq et al. [21] reported pH 7.5–8.0 to be the best for the production of alpha amylase by Bacillus subtilis. Further increase and decrease in the medium pH reduced the enzyme production, activity, and stability (Figure 4). The pH change observed during the growth of the organism also affects product stability in the medium. Results show that enzyme production was generally active and stable from pH 5.0 to 10.0, which indicates excellent buffering property. The pH values also serve as a valuable indicator of the initiation and end of enzyme synthesis [22].

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