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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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Phylogenetic tree showing relation between strain RG-01 and other Bacillus strains.
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Related In: Results  -  Collection


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fig1: Phylogenetic tree showing relation between strain RG-01 and other Bacillus strains.

Mentions: The efficient strain RG-01 was rod-shaped, Gram-positive, aerobe and facultative, motile, with positive Voges-Proskauer, catalase and oxidase test. It grew over a wide range of pH (5–12), temperature (15°–85°C), and NaCl concentration (0.0–16%), and it was able to hydrolyze gelatin, esculin, starch, Tween-20, and Tween-40 and produce acid from glucose. The strain was halotolerant as it grew in the presence of 0.0–12% NaCl but did not require salt for its physiological activities (Table 1). On account of morphological and biochemical characteristics, it was identified as Bacillus sp. by MTCC MTECH, Chandigarh, India. Analysis of 16S rDNA sequence (1444 bp) revealed its 99.3% homology with Bacillus tequilensis strains and was designated as Bacillus tequilensis RG-09. The 1444 bp 16S rDNA sequence was submitted to GenBank (JQ: 619484). The strain RG-09 was in the same cluster of phylogenetic tree (Figure 1) with different strains of Bacillus tequilensis. However, the 16S rDNA sequence analysis indicates that it is a different and novel strain of Bacillus tequilensis. The 16S rDNA is the most widely accepted gene employed for bacterial classification and identification. Gupta et al. [16] emphasized that the use of molecular markers like 16S rDNA as species-specific identification tool has been provided with a truly “microscopic” sensitivity down to single-cell detection.


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)

Phylogenetic tree showing relation between strain RG-01 and other Bacillus strains.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig1: Phylogenetic tree showing relation between strain RG-01 and other Bacillus strains.
Mentions: The efficient strain RG-01 was rod-shaped, Gram-positive, aerobe and facultative, motile, with positive Voges-Proskauer, catalase and oxidase test. It grew over a wide range of pH (5–12), temperature (15°–85°C), and NaCl concentration (0.0–16%), and it was able to hydrolyze gelatin, esculin, starch, Tween-20, and Tween-40 and produce acid from glucose. The strain was halotolerant as it grew in the presence of 0.0–12% NaCl but did not require salt for its physiological activities (Table 1). On account of morphological and biochemical characteristics, it was identified as Bacillus sp. by MTCC MTECH, Chandigarh, India. Analysis of 16S rDNA sequence (1444 bp) revealed its 99.3% homology with Bacillus tequilensis strains and was designated as Bacillus tequilensis RG-09. The 1444 bp 16S rDNA sequence was submitted to GenBank (JQ: 619484). The strain RG-09 was in the same cluster of phylogenetic tree (Figure 1) with different strains of Bacillus tequilensis. However, the 16S rDNA sequence analysis indicates that it is a different and novel strain of Bacillus tequilensis. The 16S rDNA is the most widely accepted gene employed for bacterial classification and identification. Gupta et al. [16] emphasized that the use of molecular markers like 16S rDNA as species-specific identification tool has been provided with a truly “microscopic” sensitivity down to single-cell detection.

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