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Molecular Identification of a Newly Isolated Bacillus subtilis BI19 and Optimization of Production Conditions for Enhanced Production of Extracellular Amylase.

Dash BK, Rahman MM, Sarker PK - Biomed Res Int (2015)

Bottom Line: Rice flour (1.25%) as a cheap natural carbon source was found to induce amylase production mostly.Addition of surfactants like Tween 80 (0.25 g/L) and sodium lauryl sulfate (0.2 g/L) resulted in 28% and 15% increase in enzyme production, respectively.These results suggest that B. subtilis BI19 could be exploited for production of amylase at relatively low cost and time.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology and Genetic Engineering, Faculty of Applied Science and Technology, Islamic University, Kushtia 7003, Bangladesh ; Department of Genetic Engineering and Biotechnology, Faculty of Biological Science and Technology, Jessore University of Science and Technology, Jessore 7408, Bangladesh.

ABSTRACT
A study was carried out with a newly isolated bacterial strain yielding extracellular amylase. The phylogenetic tree constructed on the basis of 16S rDNA gene sequences revealed this strain as clustered with the closest members of Bacillus sp. and identified as Bacillus subtilis BI19. The effect of various fermentation conditions on amylase production through shake-flask culture was investigated. Rice flour (1.25%) as a cheap natural carbon source was found to induce amylase production mostly. A combination of peptone and tryptone as organic and ammonium sulfate as inorganic nitrogen sources gave highest yield. Maximum production was obtained after 24 h of incubation at 37 °C with an initial medium pH 8.0. Addition of surfactants like Tween 80 (0.25 g/L) and sodium lauryl sulfate (0.2 g/L) resulted in 28% and 15% increase in enzyme production, respectively. Amylase production was 3.06 times higher when optimized production conditions were used. Optimum reaction temperature and pH for crude amylase activity were 50 °C and 6.0, respectively. The crude enzyme showed activity and stability over a fair range of temperature and pH. These results suggest that B. subtilis BI19 could be exploited for production of amylase at relatively low cost and time.

No MeSH data available.


Related in: MedlinePlus

The production of amylase by Bacillus subtilis BI19 in shake flask fermentation. (a) Effect of different carbon sources and (b) effect of different concentrations of rice flour. Each value is an average of three replicates, pH 7.0, incubation period 72 h, incubation temperature 37°C, agitation speed 150 rpm, and inoculums volume 2%.
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Related In: Results  -  Collection


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fig1: The production of amylase by Bacillus subtilis BI19 in shake flask fermentation. (a) Effect of different carbon sources and (b) effect of different concentrations of rice flour. Each value is an average of three replicates, pH 7.0, incubation period 72 h, incubation temperature 37°C, agitation speed 150 rpm, and inoculums volume 2%.

Mentions: The production of amylase by B. subtilis reported to be effected by various carbon sources [5]. In our study, rice flour, starch and corn flour were found to be the stimulator of amylase production (Figure 1(a)). It may be due to starch and rice flour metabolized slowly by the bacterium as complex carbohydrate sources with increasing accumulation of inducible amylase in the fermentation medium [25]. Rice flour at a concentration of 1.25% (w/v) supported optimal enzyme production, followed by a decline at higher concentrations (Figure 1(b)). This can be attributed to the high viscosity of culture broth at such concentrations that interferes with O2 transfer leading to limitations of dissolved O2 for growth of bacteria [26]. Hence, these starch-rich rice and corn flours may prove useful and cheaper alternative natural sources of carbon and energy for the bacterial production of amylase.


Molecular Identification of a Newly Isolated Bacillus subtilis BI19 and Optimization of Production Conditions for Enhanced Production of Extracellular Amylase.

Dash BK, Rahman MM, Sarker PK - Biomed Res Int (2015)

The production of amylase by Bacillus subtilis BI19 in shake flask fermentation. (a) Effect of different carbon sources and (b) effect of different concentrations of rice flour. Each value is an average of three replicates, pH 7.0, incubation period 72 h, incubation temperature 37°C, agitation speed 150 rpm, and inoculums volume 2%.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4477212&req=5

fig1: The production of amylase by Bacillus subtilis BI19 in shake flask fermentation. (a) Effect of different carbon sources and (b) effect of different concentrations of rice flour. Each value is an average of three replicates, pH 7.0, incubation period 72 h, incubation temperature 37°C, agitation speed 150 rpm, and inoculums volume 2%.
Mentions: The production of amylase by B. subtilis reported to be effected by various carbon sources [5]. In our study, rice flour, starch and corn flour were found to be the stimulator of amylase production (Figure 1(a)). It may be due to starch and rice flour metabolized slowly by the bacterium as complex carbohydrate sources with increasing accumulation of inducible amylase in the fermentation medium [25]. Rice flour at a concentration of 1.25% (w/v) supported optimal enzyme production, followed by a decline at higher concentrations (Figure 1(b)). This can be attributed to the high viscosity of culture broth at such concentrations that interferes with O2 transfer leading to limitations of dissolved O2 for growth of bacteria [26]. Hence, these starch-rich rice and corn flours may prove useful and cheaper alternative natural sources of carbon and energy for the bacterial production of amylase.

Bottom Line: Rice flour (1.25%) as a cheap natural carbon source was found to induce amylase production mostly.Addition of surfactants like Tween 80 (0.25 g/L) and sodium lauryl sulfate (0.2 g/L) resulted in 28% and 15% increase in enzyme production, respectively.These results suggest that B. subtilis BI19 could be exploited for production of amylase at relatively low cost and time.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology and Genetic Engineering, Faculty of Applied Science and Technology, Islamic University, Kushtia 7003, Bangladesh ; Department of Genetic Engineering and Biotechnology, Faculty of Biological Science and Technology, Jessore University of Science and Technology, Jessore 7408, Bangladesh.

ABSTRACT
A study was carried out with a newly isolated bacterial strain yielding extracellular amylase. The phylogenetic tree constructed on the basis of 16S rDNA gene sequences revealed this strain as clustered with the closest members of Bacillus sp. and identified as Bacillus subtilis BI19. The effect of various fermentation conditions on amylase production through shake-flask culture was investigated. Rice flour (1.25%) as a cheap natural carbon source was found to induce amylase production mostly. A combination of peptone and tryptone as organic and ammonium sulfate as inorganic nitrogen sources gave highest yield. Maximum production was obtained after 24 h of incubation at 37 °C with an initial medium pH 8.0. Addition of surfactants like Tween 80 (0.25 g/L) and sodium lauryl sulfate (0.2 g/L) resulted in 28% and 15% increase in enzyme production, respectively. Amylase production was 3.06 times higher when optimized production conditions were used. Optimum reaction temperature and pH for crude amylase activity were 50 °C and 6.0, respectively. The crude enzyme showed activity and stability over a fair range of temperature and pH. These results suggest that B. subtilis BI19 could be exploited for production of amylase at relatively low cost and time.

No MeSH data available.


Related in: MedlinePlus