An extracellular antifungal chitinase from Lecanicillium lecanii: purification, properties, and application in biocontrol against plant pathogenic fungi

Huu Quan NGUYEN; Dinh Thi QUYEN; Sy Le Thanh NGUYEN; Van Hanh VU

doi:https://doi.org/10.3906/biy-1402-28

Makale detayları

An extracellular antifungal chitinase from Lecanicillium lecanii: purification, properties, and application in biocontrol against plant pathogenic fungi

Yazarlar:

Huu Quan NGUYEN | Dinh Thi QUYEN | Sy Le Thanh NGUYEN | Van Hanh VU

Yayıncı:

TUBITAK

Ana Başlıklar:

Yaşam bilimleri

Dergi:

Turkish Journal of Biology 2015 Vol.39 Issue 1, pp.6-14

Belge Tipi:

Makale

Yayın Tarihi:

3/2015

Belge Dili:

İngilizce

Document ID (Org No.):

2000812008

Sıklık:

İki Aylık

eISSN:

1303-6092

pISSN:

1300-0152

DOI:

https://doi.org/10.3906/biy-1402-28

Anahtar kelimeler:

Özet:

 

An extracellular antifungal chitinase from L. lecanii strain 43H was purified by ammonium sulfate precipitation and DEAE-Sephadex A-50 ion exchange chromatography; it showed a molecular mass of approximately 33 kDa with a specific activity of 167.5 U/mg protein and purification factor of 2.5. Optimum temperature and pH were observed at 40 °C and pH 6.0, respectively. This enzyme was stable at up to 40 °C and at pH 5.0-6.0. The kinetic constants Km and Vmax determined for the chitinase with colloidal chitin as substrate was 0.82 mg/mL and 4.51 U/mg, respectively. The presence of 5-15 mM tested metal ions led to activation of the chitinase activity with an increase of up to 126% except for Al3+, Ag+, and Hg2+. Tween 80 (0.5%), Tween 20 (1%-2%), and Triton X-100 (1%) increased the enzyme activity by up to 25%, whereas higher concentration of 2% SDS completely inhibited the enzyme. The chitinase exhibited high resistance to organic solvents (methanol, acetone) and retained 89%-95% of its initial activity. The chitinase was found to inhibit the conidial germination and mycelial growth of plant pathogenic fungi. These results suggest that chitinase might be used in enzymatic reactions and as a potential fungicide against pathogens.

 

An extracellular antifungal chitinase from L. lecanii strain 43H was purified by ammonium sulfate precipitation and DEAE-Sephadex A-50 ion exchange chromatography; it showed a molecular mass of approximately 33 kDa with a specific activity of 167.5 U/mg protein and purification factor of 2.5. Optimum temperature and pH were observed at 40 °C and pH 6.0, respectively. This enzyme was stable at up to 40 °C and at pH 5.0-6.0. The kinetic constants Km and Vmax determined for the chitinase with colloidal chitin as substrate was 0.82 mg/mL and 4.51 U/mg, respectively. The presence of 5-15 mM tested metal ions led to activation of the chitinase activity with an increase of up to 126% except for Al3+, Ag+, and Hg2+. Tween 80 (0.5%), Tween 20 (1%-2%), and Triton X-100 (1%) increased the enzyme activity by up to 25%, whereas higher concentration of 2% SDS completely inhibited the enzyme. The chitinase exhibited high resistance to organic solvents (methanol, acetone) and retained 89%-95% of its initial activity. The chitinase was found to inhibit the conidial germination and mycelial growth of plant pathogenic fungi. These results suggest that chitinase might be used in enzymatic reactions and as a potential fungicide against pathogens.

Makale detayları

An extracellular antifungal chitinase from Lecanicillium lecanii: purification, properties, and application in biocontrol against plant pathogenic fungi

Yazarlar:

Huu Quan NGUYEN | Dinh Thi QUYEN | Sy Le Thanh NGUYEN | Van Hanh VU

Yayıncı:

TUBITAK

Ana Başlıklar:

Yaşam bilimleri

Dergi:

Turkish Journal of Biology 2015 Vol.39 Issue 1, pp.6-14

Belge Tipi:

Makale

Yayın Tarihi:

3/2015

Belge Dili:

İngilizce

Document ID (Org No.):

2000812008

Sıklık:

İki Aylık

eISSN:

1303-6092

pISSN:

1300-0152

DOI:

https://doi.org/10.3906/biy-1402-28

Antifungal activity

characterization

chitinase

Lecanicillium lecanii

purification

Sayı

Makale detayları

An extracellular antifungal chitinase from Lecanicillium lecanii: purification, properties, and application in biocontrol against plant pathogenic fungi

Yazarlar: Huu Quan NGUYEN | Dinh Thi QUYEN | Sy Le Thanh NGUYEN | Van Hanh VU

Yayıncı: TUBITAK

Ana Başlıklar: Yaşam bilimleri

Dergi: Turkish Journal of Biology 2015 Vol.39 Issue 1, pp.6-14

Belge Tipi: Makale

Yayın Tarihi: 3/2015

Belge Dili: İngilizce

Document ID (Org No.): 2000812008

Sıklık: İki Aylık

eISSN: 1303-6092

pISSN: 1300-0152

DOI: https://doi.org/10.3906/biy-1402-28

Antifungal activity

characterization

chitinase

Lecanicillium lecanii

purification

Sayı

Yazarlar:

Huu Quan NGUYEN | Dinh Thi QUYEN | Sy Le Thanh NGUYEN | Van Hanh VU

Yayıncı:

TUBITAK

Ana Başlıklar:

Yaşam bilimleri

Dergi:

Turkish Journal of Biology 2015 Vol.39 Issue 1, pp.6-14

Belge Tipi:

Makale

Yayın Tarihi:

3/2015

Belge Dili:

İngilizce

Document ID (Org No.):

2000812008

Sıklık:

İki Aylık

eISSN:

1303-6092

pISSN:

1300-0152

DOI:

https://doi.org/10.3906/biy-1402-28