Production, purifi cation, and characterization of α-amylase by Bacillus subtilis and its mutant derivates

Elif DEMİRKAN

Article Details

Production, purifi cation, and characterization of α-amylase by Bacillus subtilis and its mutant derivates

Authors:

Elif DEMİRKAN

Publisher:

TUBITAK

Main Topics:

Life sciences

Journal:

Turkish Journal of Biology 2011 Vol.35 Issue 6, pp.705-712

Document Type:

Article

Publication Date:

5/2011

Document Language:

English

Document ID (Org No.):

2000812753

Frequency:

Bimonthly

eISSN:

1303-6092

pISSN:

1300-0152

Keywords:

Abstract:

 

The effects of various carbon and nitrogen sources on production of a-amylase by Bacillus subtilis and its mutant derivates were investigated. The maximum production of a-amylase by all strains was obtained in the presence of mesoinositol as the carbon source. There was no more significant increase in enzyme yield in the case of the supplementation of nitrogen sources, whereas malt extract and tryptone were preferred nitrogen sources for amylase production by Bacillus subtilis and mutant U 2-6 strain, respectively. a-Amylases of B. subtilis and its mutant strain (EBUE 5-3) were purified through a series of steps, and characterized. The optimum temperature and pH values of the purified amylases were found to be 45 °C and 6.0, respectively. The enzyme of mutant strain had more stability than the enzyme of the parental strain in alkaline conditions (85% at pH 8.0 for 1 h). The Km and Vmax values of both amylases were also compared. Enzymes were strongly inhibited by Cu2+, Hg2+, and Ag2+, but activated by Ca2+, Ba2+, Mg2+, Li2+, and Mn2+. Metal ion concentration of 1 mM had a greater effect on enzyme activities than 5 mM did. The estimated molecular weight of the purified enzymes was 56 kDa. The N-terminal amino acid sequence of amylases produced by the parental and the mutant strain showed homology.

 

The effects of various carbon and nitrogen sources on production of a-amylase by Bacillus subtilis and its mutant derivates were investigated. The maximum production of a-amylase by all strains was obtained in the presence of mesoinositol as the carbon source. There was no more significant increase in enzyme yield in the case of the supplementation of nitrogen sources, whereas malt extract and tryptone were preferred nitrogen sources for amylase production by Bacillus subtilis and mutant U 2-6 strain, respectively. a-Amylases of B. subtilis and its mutant strain (EBUE 5-3) were purified through a series of steps, and characterized. The optimum temperature and pH values of the purified amylases were found to be 45 °C and 6.0, respectively. The enzyme of mutant strain had more stability than the enzyme of the parental strain in alkaline conditions (85% at pH 8.0 for 1 h). The Km and Vmax values of both amylases were also compared. Enzymes were strongly inhibited by Cu2+, Hg2+, and Ag2+, but activated by Ca2+, Ba2+, Mg2+, Li2+, and Mn2+. Metal ion concentration of 1 mM had a greater effect on enzyme activities than 5 mM did. The estimated molecular weight of the purified enzymes was 56 kDa. The N-terminal amino acid sequence of amylases produced by the parental and the mutant strain showed homology.

Article Details

Production, purifi cation, and characterization of α-amylase by Bacillus subtilis and its mutant derivates

Authors:

Elif DEMİRKAN

Publisher:

TUBITAK

Main Topics:

Life sciences

Journal:

Turkish Journal of Biology 2011 Vol.35 Issue 6, pp.705-712

Document Type:

Article

Publication Date:

5/2011

Document Language:

English

Document ID (Org No.):

2000812753

Frequency:

Bimonthly

eISSN:

1303-6092

pISSN:

1300-0152

Key words: Bacillus subtilis

mutant

a-amylase

production

purification

characterization

Issues

Article Details

Production, purifi cation, and characterization of α-amylase by Bacillus subtilis and its mutant derivates

Authors: Elif DEMİRKAN

Publisher: TUBITAK

Main Topics: Life sciences

Journal: Turkish Journal of Biology 2011 Vol.35 Issue 6, pp.705-712

Document Type: Article

Publication Date: 5/2011

Document Language: English

Document ID (Org No.): 2000812753

Frequency: Bimonthly

eISSN: 1303-6092

pISSN: 1300-0152

Key words: Bacillus subtilis

mutant

a-amylase

production

purification

characterization

Issues

Authors:

Elif DEMİRKAN

Publisher:

TUBITAK

Main Topics:

Life sciences

Journal:

Turkish Journal of Biology 2011 Vol.35 Issue 6, pp.705-712

Document Type:

Article

Publication Date:

5/2011

Document Language:

English

Document ID (Org No.):

2000812753

Frequency:

Bimonthly

eISSN:

1303-6092

pISSN:

1300-0152