Purification and biochemical characterization of an extracellular lipase from psychrotolerant Pseudomonas fluorescens KE38

Aysun ADAN GÖKBULUT; Alper ARSLANOĞLU

Makale detayları

Purification and biochemical characterization of an extracellular lipase from psychrotolerant Pseudomonas fluorescens KE38

Yazarlar:

Aysun ADAN GÖKBULUT | Alper ARSLANOĞLU

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Biology 2013 Vol.37 Issue 5, pp.538-546

Belge Tipi:

Makale

Yayın Tarihi:

4/2013

Belge Dili:

Türkçe

Document ID (Org No.):

2000812354

Sıklık:

İki Aylık

eISSN:

1303-6092

pISSN:

1300-0152

Anahtar kelimeler:

Özet:

 

An extracellular lipase producing bacterium was isolated from a soil sample, and identified as a strain of Pseudomonas fluorescens by 16S rRNA gene sequencing. It was named Pseudomonas fluorescens KE38. KE38 showed psychrotolerant properties with an optimum growth temperature of 25 °C. The lipase enzyme secreted by KE38 was purified 41.13-fold with an overall yield of 54.99%, and a specific activity of 337.3 U/mg. The molecular mass of purified lipase was estimated to be approximately 43 kDa by SDS-PAGE. Although the lipase was active at a temperature range of 1565 °C, it exhibited maximum activity at 45 °C, at pH 8.0. The enzyme exhibited high stability retaining 100% and 70% of its activity after an incubation period of 45 and 100 min at 45 °C and pH 8.0 respectively. It also showed a broad substrate specificity acting on p-nitrophenyl esters with C8-C18 acyl groups as substrates and was activated by Ca2+ and Ni2+ at 1 mM. While the enzyme retained its activity levels in the presence of a variety of organic solvents, DMSO and dimethylformamide enhanced this. High stability, broad substrate specificity and activity at cold temperatures in the presence of organic solvents, and metal ions make the extracellular lipase of KE38 a candidate for industrial applications.

 

An extracellular lipase producing bacterium was isolated from a soil sample, and identified as a strain of Pseudomonas fluorescens by 16S rRNA gene sequencing. It was named Pseudomonas fluorescens KE38. KE38 showed psychrotolerant properties with an optimum growth temperature of 25 °C. The lipase enzyme secreted by KE38 was purified 41.13-fold with an overall yield of 54.99%, and a specific activity of 337.3 U/mg. The molecular mass of purified lipase was estimated to be approximately 43 kDa by SDS-PAGE. Although the lipase was active at a temperature range of 1565 °C, it exhibited maximum activity at 45 °C, at pH 8.0. The enzyme exhibited high stability retaining 100% and 70% of its activity after an incubation period of 45 and 100 min at 45 °C and pH 8.0 respectively. It also showed a broad substrate specificity acting on p-nitrophenyl esters with C8-C18 acyl groups as substrates and was activated by Ca2+ and Ni2+ at 1 mM. While the enzyme retained its activity levels in the presence of a variety of organic solvents, DMSO and dimethylformamide enhanced this. High stability, broad substrate specificity and activity at cold temperatures in the presence of organic solvents, and metal ions make the extracellular lipase of KE38 a candidate for industrial applications.

Makale detayları

Purification and biochemical characterization of an extracellular lipase from psychrotolerant Pseudomonas fluorescens KE38

Yazarlar:

Aysun ADAN GÖKBULUT | Alper ARSLANOĞLU

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Biology 2013 Vol.37 Issue 5, pp.538-546

Belge Tipi:

Makale

Yayın Tarihi:

4/2013

Belge Dili:

Türkçe

Document ID (Org No.):

2000812354

Sıklık:

İki Aylık

eISSN:

1303-6092

pISSN:

1300-0152

Key words: Pseudomonas fluorescens

extracellular lipase

enzyme purification

Sayı

Makale detayları

Purification and biochemical characterization of an extracellular lipase from psychrotolerant Pseudomonas fluorescens KE38

Yazarlar: Aysun ADAN GÖKBULUT | Alper ARSLANOĞLU

Yayıncı: TUBITAK

Dergi: Turkish Journal of Biology 2013 Vol.37 Issue 5, pp.538-546

Belge Tipi: Makale

Yayın Tarihi: 4/2013

Belge Dili: Türkçe

Document ID (Org No.): 2000812354

Sıklık: İki Aylık

eISSN: 1303-6092

pISSN: 1300-0152

Key words: Pseudomonas fluorescens

extracellular lipase

enzyme purification

Sayı

Yazarlar:

Aysun ADAN GÖKBULUT | Alper ARSLANOĞLU

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Biology 2013 Vol.37 Issue 5, pp.538-546

Belge Tipi:

Makale

Yayın Tarihi:

4/2013

Belge Dili:

Türkçe

Document ID (Org No.):

2000812354

Sıklık:

İki Aylık

eISSN:

1303-6092

pISSN:

1300-0152