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

Elif DEMİRKAN

Makale detayları

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

Yazarlar:

Elif DEMİRKAN

Yayıncı:

TUBITAK

Ana Başlıklar:

Yaşam bilimleri

Dergi:

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

Belge Tipi:

Makale

Yayın Tarihi:

5/2011

Belge Dili:

İngilizce

Document ID (Org No.):

2000812753

Sıklık:

İki Aylık

eISSN:

1303-6092

pISSN:

1300-0152

Anahtar kelimeler:

Özet:

 

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.

Makale detayları

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

Yazarlar:

Elif DEMİRKAN

Yayıncı:

TUBITAK

Ana Başlıklar:

Yaşam bilimleri

Dergi:

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

Belge Tipi:

Makale

Yayın Tarihi:

5/2011

Belge Dili:

İngilizce

Document ID (Org No.):

2000812753

Sıklık:

İki Aylık

eISSN:

1303-6092

pISSN:

1300-0152

Key words: Bacillus subtilis

mutant

a-amylase

production

purification

characterization

Sayı

Makale detayları

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

Yazarlar: Elif DEMİRKAN

Yayıncı: TUBITAK

Ana Başlıklar: Yaşam bilimleri

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

Belge Tipi: Makale

Yayın Tarihi: 5/2011

Belge Dili: İngilizce

Document ID (Org No.): 2000812753

Sıklık: İki Aylık

eISSN: 1303-6092

pISSN: 1300-0152

Key words: Bacillus subtilis

mutant

a-amylase

production

purification

characterization

Sayı

Yazarlar:

Elif DEMİRKAN

Yayıncı:

TUBITAK

Ana Başlıklar:

Yaşam bilimleri

Dergi:

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

Belge Tipi:

Makale

Yayın Tarihi:

5/2011

Belge Dili:

İngilizce

Document ID (Org No.):

2000812753

Sıklık:

İki Aylık

eISSN:

1303-6092

pISSN:

1300-0152