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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 different metal ions on amylase production and stability. The control flask does not contain any metal ions. Test flasks contained different metal ions in the medium at a level of 25 mM. The flasks were inoculated with culture and were incubated at initial pH 7.0, 55°C for 24 h. For enzyme activity, the reaction was assayed and for stability enzyme was preincubated with different metal ions (25 mM) 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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fig10: Effect of different metal ions on amylase production and stability. The control flask does not contain any metal ions. Test flasks contained different metal ions in the medium at a level of 25 mM. The flasks were inoculated with culture and were incubated at initial pH 7.0, 55°C for 24 h. For enzyme activity, the reaction was assayed and for stability enzyme was preincubated with different metal ions (25 mM) 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: For the study of the effect of metal ions, viz, NaCl, CaCl2, MgSO4, HgCl2, FeSO4, NiCl2, CuSO4, CoCl2, and ZnCl2 at a concentration of 25 mM were individually tested in the basal medium at their optimal temperature, incubation period, and pH to observe the effect on enzyme production and stability by Bacillus tequilensis RG-01. Out of these metal ions, sodium and calcium ions were found to be the best for amylase production (6500 and 6750 U/mL) and stability (120 and 145%). While amylase production and stability of Bacillus tequilensis RG-01 were slightly reduced in the presence of Fe2+, Mg2+, Cu2+, Ni2-, Co2+, and Zn2+. Bacillus tequilensis RG-01 also tolerates 25 mM HgCl2, a novel finding of this strain (Figure 10). Amylolytic enzymes are metalloenzymes with up to six Ca2+ atoms at the active site whose catalytic activity can be affected by mono- and divalent metals [35]. Ca2+ enhanced the activity of α-amylase from C. perfringens [36], B. subtilis [37], and Acremonium sporosulcatum [38]. The inhibition of B. subtilis JS-2004 α-amylase by Co2+, Cu2+, and Ba2+ ions could be due to competition between the exogenous cations and the protein associated cations, resulting in decreased metalloenzymes activity [39]. Hernandez et al. [40] reported that amylase activity was strongly inhibited by Cu2+, Hg2+, and Zn2+.


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 different metal ions on amylase production and stability. The control flask does not contain any metal ions. Test flasks contained different metal ions in the medium at a level of 25 mM. The flasks were inoculated with culture and were incubated at initial pH 7.0, 55°C for 24 h. For enzyme activity, the reaction was assayed and for stability enzyme was preincubated with different metal ions (25 mM) 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

fig10: Effect of different metal ions on amylase production and stability. The control flask does not contain any metal ions. Test flasks contained different metal ions in the medium at a level of 25 mM. The flasks were inoculated with culture and were incubated at initial pH 7.0, 55°C for 24 h. For enzyme activity, the reaction was assayed and for stability enzyme was preincubated with different metal ions (25 mM) 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: For the study of the effect of metal ions, viz, NaCl, CaCl2, MgSO4, HgCl2, FeSO4, NiCl2, CuSO4, CoCl2, and ZnCl2 at a concentration of 25 mM were individually tested in the basal medium at their optimal temperature, incubation period, and pH to observe the effect on enzyme production and stability by Bacillus tequilensis RG-01. Out of these metal ions, sodium and calcium ions were found to be the best for amylase production (6500 and 6750 U/mL) and stability (120 and 145%). While amylase production and stability of Bacillus tequilensis RG-01 were slightly reduced in the presence of Fe2+, Mg2+, Cu2+, Ni2-, Co2+, and Zn2+. Bacillus tequilensis RG-01 also tolerates 25 mM HgCl2, a novel finding of this strain (Figure 10). Amylolytic enzymes are metalloenzymes with up to six Ca2+ atoms at the active site whose catalytic activity can be affected by mono- and divalent metals [35]. Ca2+ enhanced the activity of α-amylase from C. perfringens [36], B. subtilis [37], and Acremonium sporosulcatum [38]. The inhibition of B. subtilis JS-2004 α-amylase by Co2+, Cu2+, and Ba2+ ions could be due to competition between the exogenous cations and the protein associated cations, resulting in decreased metalloenzymes activity [39]. Hernandez et al. [40] reported that amylase activity was strongly inhibited by Cu2+, Hg2+, and Zn2+.

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