Immobilized metal ion affinity nanospheres for a-amylase immobilization

Tülden KALBURCU; Münire Nalan TÜZMEN

Makale detayları

Immobilized metal ion affinity nanospheres for a-amylase immobilization

Yazarlar:

Tülden KALBURCU | Münire Nalan TÜZMEN

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Chemistry 2014 Vol.38 Issue 1, pp.28-40

Belge Tipi:

Makale

Yayın Tarihi:

5/2014

Belge Dili:

Türkçe

Document ID (Org No.):

2000727238

Sıklık:

İki Aylık

eISSN:

1303-6130

pISSN:

1300-0527

Özet:

 

Immobilized metal chelate affinity chromatography (IMAC) support was practiced for a-amylase immobilization. Poly(hydroxyethylmethacrylate-methacryloylamidotryptophan)-Ni2+ [p(HEMA-MAT)-Ni2+] nanospheres, average diameter 100 nm, were produced by surfactant free emulsion polymerization. Characterizations of p(HEMA-MAT)-Ni2+ nanospheres were carried out by Fourier transform infrared (FTIR) spectroscopy and scanning electron microscope (SEM). In addition, average particle size, size distribution, and surface charge were specified. The amount of N-methacryloylamidotryptophan (MAT) incorporated to polymer was determined as 1.95 mmol/g polymers by using nitrogen stoichiometry. The specific surface areas of poly(hydroxyethylmethacrylate) [p(HEMA)] and p(HEMA-MAT) nanospheres were calculated as 1856 m2/g and 1914 m2/g, respectively. Protein adsorption increased with increasing initial protein concentration and maximum a-amylase adsorption on p(HEMA-MAT)-Ni2+ nanospheres was observed at pH 4.0. Both free and immobilized a-amylase showed pH optimum at pH 7.0. It was determined that the immobilized a-amylase had better thermostability than the free one. Immobilization of the enzyme did not significantly change the kinetic parameters. The storage stability of a-amylase increased upon immobilization. It was also observed that p(HEMA-MAT)-Ni2+ nanospheres can be repeatedly used for a-amylase immobilization.

 

Immobilized metal chelate affinity chromatography (IMAC) support was practiced for a-amylase immobilization. Poly(hydroxyethylmethacrylate-methacryloylamidotryptophan)-Ni2+ [p(HEMA-MAT)-Ni2+] nanospheres, average diameter 100 nm, were produced by surfactant free emulsion polymerization. Characterizations of p(HEMA-MAT)-Ni2+ nanospheres were carried out by Fourier transform infrared (FTIR) spectroscopy and scanning electron microscope (SEM). In addition, average particle size, size distribution, and surface charge were specified. The amount of N-methacryloylamidotryptophan (MAT) incorporated to polymer was determined as 1.95 mmol/g polymers by using nitrogen stoichiometry. The specific surface areas of poly(hydroxyethylmethacrylate) [p(HEMA)] and p(HEMA-MAT) nanospheres were calculated as 1856 m2/g and 1914 m2/g, respectively. Protein adsorption increased with increasing initial protein concentration and maximum a-amylase adsorption on p(HEMA-MAT)-Ni2+ nanospheres was observed at pH 4.0. Both free and immobilized a-amylase showed pH optimum at pH 7.0. It was determined that the immobilized a-amylase had better thermostability than the free one. Immobilization of the enzyme did not significantly change the kinetic parameters. The storage stability of a-amylase increased upon immobilization. It was also observed that p(HEMA-MAT)-Ni2+ nanospheres can be repeatedly used for a-amylase immobilization.

Makale detayları

Immobilized metal ion affinity nanospheres for a-amylase immobilization

Yazarlar: Tülden KALBURCU | Münire Nalan TÜZMEN

Yayıncı: TUBITAK

Dergi: Turkish Journal of Chemistry 2014 Vol.38 Issue 1, pp.28-40

Belge Tipi: Makale

Yayın Tarihi: 5/2014

Belge Dili: Türkçe

Document ID (Org No.): 2000727238

Sıklık: İki Aylık

eISSN: 1303-6130

pISSN: 1300-0527

Sayı

Yazarlar:

Tülden KALBURCU | Münire Nalan TÜZMEN

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Chemistry 2014 Vol.38 Issue 1, pp.28-40

Belge Tipi:

Makale

Yayın Tarihi:

5/2014

Belge Dili:

Türkçe

Document ID (Org No.):

2000727238

Sıklık:

İki Aylık

eISSN:

1303-6130

pISSN:

1300-0527