Production and Characterization of crude chitinase from Trichoderma virens

Chitinase hydrolyzes chitin, a biopolymer of N-acetylglucosamine which is being widely used in biological and agricultural research. Optimization of chitinase production by Trichoderma virens was conducted using batch cultures of Trichoderma Minimal Medium (TM medium) that consists of 2.0% (w/v) col...

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Main Authors: Wasli, Azaliza Safarida, Md Salleh, Madihah, Md Illias, Rosli
Format: Article
Language:English
Published: School of Postgraduate Studies, UTM 2006
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Online Access:http://eprints.utm.my/id/eprint/3045/1/RPCES_2006_Dr_Madihah.pdf
http://eprints.utm.my/id/eprint/3045/
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Institution: Universiti Teknologi Malaysia
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spelling my.utm.30452017-10-24T01:04:48Z http://eprints.utm.my/id/eprint/3045/ Production and Characterization of crude chitinase from Trichoderma virens Wasli, Azaliza Safarida Md Salleh, Madihah Md Illias, Rosli Q Science (General) Chitinase hydrolyzes chitin, a biopolymer of N-acetylglucosamine which is being widely used in biological and agricultural research. Optimization of chitinase production by Trichoderma virens was conducted using batch cultures of Trichoderma Minimal Medium (TM medium) that consists of 2.0% (w/v) colloidal chitin as substrate. Fermentation was performed at agitation speed of 200 rpm and incubated at 30ºC in 250 ml shake flasks. Optimum chitin concentration was obtained at 4.0% (w/v). The addition of a stimulant, methanol, at 0.2% (v/v) enhances chitinase production by 71.4%. Higher concentrations of methanol would inhibit chitinase production. Works for optimizing agitation speed was carried out in a 2.0 L batch culture using B.Braun Biostat-B bioreactor. The optimum conditions for chitinase productions were performed in the same bioreactor using disc turbine impellers (diameter of 5.5 cm) with air flow rate of 1.0 l/m and agitation speed of 250 rpm resulting in an increased chitinase activity of 0.447 U/ml compared to 0.052 U/ml in shake flask fermentation. This improvement of 88.4% might be due to efficient mixing and high oxygen transfer rate which is essential for fungal growth. Characterization of crude chitinase was carried out to determine the optimal conditions for the crude chitinase activity to hydrolyze chitin and the crude enzyme stability. The crucial physical factors affecting the optimum and stability of chitinase activity were pH and temperature. pH optimum was achieved at pH 4.0 in citrate phosphate buffer and chitinase was very stable at pH 3.0-7.0, with 350% of residual activity is retained at pH 7.0. Optimum temperature for chitinase activity was achieved at 50ºC while its stability decreases with the increment of temperature. Only 37.5% of residual chitinase activity was retained at 70ºC. School of Postgraduate Studies, UTM 2006-07-25 Article NonPeerReviewed application/pdf en http://eprints.utm.my/id/eprint/3045/1/RPCES_2006_Dr_Madihah.pdf Wasli, Azaliza Safarida and Md Salleh, Madihah and Md Illias, Rosli (2006) Production and Characterization of crude chitinase from Trichoderma virens. Regional Postgraduate Conference on Engineering and Science . pp. 625-630.
institution Universiti Teknologi Malaysia
building UTM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Teknologi Malaysia
content_source UTM Institutional Repository
url_provider http://eprints.utm.my/
language English
topic Q Science (General)
spellingShingle Q Science (General)
Wasli, Azaliza Safarida
Md Salleh, Madihah
Md Illias, Rosli
Production and Characterization of crude chitinase from Trichoderma virens
description Chitinase hydrolyzes chitin, a biopolymer of N-acetylglucosamine which is being widely used in biological and agricultural research. Optimization of chitinase production by Trichoderma virens was conducted using batch cultures of Trichoderma Minimal Medium (TM medium) that consists of 2.0% (w/v) colloidal chitin as substrate. Fermentation was performed at agitation speed of 200 rpm and incubated at 30ºC in 250 ml shake flasks. Optimum chitin concentration was obtained at 4.0% (w/v). The addition of a stimulant, methanol, at 0.2% (v/v) enhances chitinase production by 71.4%. Higher concentrations of methanol would inhibit chitinase production. Works for optimizing agitation speed was carried out in a 2.0 L batch culture using B.Braun Biostat-B bioreactor. The optimum conditions for chitinase productions were performed in the same bioreactor using disc turbine impellers (diameter of 5.5 cm) with air flow rate of 1.0 l/m and agitation speed of 250 rpm resulting in an increased chitinase activity of 0.447 U/ml compared to 0.052 U/ml in shake flask fermentation. This improvement of 88.4% might be due to efficient mixing and high oxygen transfer rate which is essential for fungal growth. Characterization of crude chitinase was carried out to determine the optimal conditions for the crude chitinase activity to hydrolyze chitin and the crude enzyme stability. The crucial physical factors affecting the optimum and stability of chitinase activity were pH and temperature. pH optimum was achieved at pH 4.0 in citrate phosphate buffer and chitinase was very stable at pH 3.0-7.0, with 350% of residual activity is retained at pH 7.0. Optimum temperature for chitinase activity was achieved at 50ºC while its stability decreases with the increment of temperature. Only 37.5% of residual chitinase activity was retained at 70ºC.
format Article
author Wasli, Azaliza Safarida
Md Salleh, Madihah
Md Illias, Rosli
author_facet Wasli, Azaliza Safarida
Md Salleh, Madihah
Md Illias, Rosli
author_sort Wasli, Azaliza Safarida
title Production and Characterization of crude chitinase from Trichoderma virens
title_short Production and Characterization of crude chitinase from Trichoderma virens
title_full Production and Characterization of crude chitinase from Trichoderma virens
title_fullStr Production and Characterization of crude chitinase from Trichoderma virens
title_full_unstemmed Production and Characterization of crude chitinase from Trichoderma virens
title_sort production and characterization of crude chitinase from trichoderma virens
publisher School of Postgraduate Studies, UTM
publishDate 2006
url http://eprints.utm.my/id/eprint/3045/1/RPCES_2006_Dr_Madihah.pdf
http://eprints.utm.my/id/eprint/3045/
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