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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 addition of different concentrations of glucose in basal medium and (b) effect of addition of various elements in basal medium.
© Copyright Policy - open-access
Related In: Results  -  Collection


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fig4: The production of amylase by Bacillus subtilis BI19 in shake flask fermentation. (a) Effect of addition of different concentrations of glucose in basal medium and (b) effect of addition of various elements in basal medium.

Mentions: Addition of free glucose in the fermentation medium was found to suppress amylase production greatly as shown in Figure 4(a). The inhibitory effect of glucose on α-amylase synthesis increased with the increase of glucose concentration in the medium. Addition of 2% glucose resulted in about 54% of production loss (3.33 U/mL/min). Similar results were found by Nusrat and Rahman (2007) [21]. The most possible reason may the suppression of synthesis of carbohydrate degrading enzymes by readily metabolizable substrates such as glucose and fructose (mediated by the protein encoded by the CreA gene) [31].


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 addition of different concentrations of glucose in basal medium and (b) effect of addition of various elements in basal medium.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig4: The production of amylase by Bacillus subtilis BI19 in shake flask fermentation. (a) Effect of addition of different concentrations of glucose in basal medium and (b) effect of addition of various elements in basal medium.
Mentions: Addition of free glucose in the fermentation medium was found to suppress amylase production greatly as shown in Figure 4(a). The inhibitory effect of glucose on α-amylase synthesis increased with the increase of glucose concentration in the medium. Addition of 2% glucose resulted in about 54% of production loss (3.33 U/mL/min). Similar results were found by Nusrat and Rahman (2007) [21]. The most possible reason may the suppression of synthesis of carbohydrate degrading enzymes by readily metabolizable substrates such as glucose and fructose (mediated by the protein encoded by the CreA gene) [31].

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