Studies on the protein-carbohydrate interaction of the Trichoderma harzianum chitinase Chit42

Research output: Contribution to conferenceConference AbstractScientific

Abstract

Trichoderma harzianum is a filamentous fungus that can secrete several chitinases (EC 3.2.1.14) to the culture medium under inductive conditions. T. harzianum Chit42 is among the best studied Trichoderma chitinases hydrolysing polymeric chitin mainly into chitobiose, a disaccharide. Chit42 is a single catalytic module enzyme, which belongs to the glycosyl hydrolase family 18 having a (beta/alfa)8 barrel fold. A three-dimensional structural model of T. harzianum Chit42 was built based on the 53% amino acid sequence identity with another fungal chitinase CiX1 from Coccidioides immitis, whose crystal structure has been determined (1). Our modelling studies suggested that Chit42 has at least 7 subsites (-5-4-3-2-1+1+2) for binding the polymeric substrate, and the substrate binding groove is formed by loops positioned between the carboxy-terminal end of the beta-strands and the amino-terminal end of the alfa-helices (2). We have additionally shown that the wild-type Chit42 can recognize animal like, beta-1,4-galactosylated and alfa-1,3-fucosylated chito-oligosaccharides (2). Our aim is to investigate the interactions of the T. harzianum Chit42 with natural and modified chito-oligosaccharides using wild-type and different engineered chitinase variants. The work will involve heterologous expression of the chitinase cDNA in E.coli, protein engineering by rational design or by directed evolution methods, and characterising the wild-type and mutant chitinases for their activity and binding specificity. The characterisation of the protein-carbohydrate interactions will include the usage of atomic force microscopy force-distance curves.
Original languageEnglish
Number of pages1
Publication statusPublished - 2006
MoE publication typeNot Eligible
Event9th European Training Course on Carbohydrates - Wageningen, Netherlands
Duration: 6 Jun 20069 Jun 2006
Conference number: 9

Course

Course9th European Training Course on Carbohydrates
CountryNetherlands
CityWageningen
Period6/06/069/06/06

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Chitinases
Carbohydrates
Proteins
Oligosaccharides
Chitin
Disaccharides
Escherichia coli Proteins
Hydrolases
Substrates
Fungi
Culture Media
Atomic force microscopy
Animals
Complementary DNA
Crystal structure
Amino Acids
Enzymes

Cite this

@conference{e47a81cfc76140008eadaf756e3de953,
title = "Studies on the protein-carbohydrate interaction of the Trichoderma harzianum chitinase Chit42",
abstract = "Trichoderma harzianum is a filamentous fungus that can secrete several chitinases (EC 3.2.1.14) to the culture medium under inductive conditions. T. harzianum Chit42 is among the best studied Trichoderma chitinases hydrolysing polymeric chitin mainly into chitobiose, a disaccharide. Chit42 is a single catalytic module enzyme, which belongs to the glycosyl hydrolase family 18 having a (beta/alfa)8 barrel fold. A three-dimensional structural model of T. harzianum Chit42 was built based on the 53{\%} amino acid sequence identity with another fungal chitinase CiX1 from Coccidioides immitis, whose crystal structure has been determined (1). Our modelling studies suggested that Chit42 has at least 7 subsites (-5-4-3-2-1+1+2) for binding the polymeric substrate, and the substrate binding groove is formed by loops positioned between the carboxy-terminal end of the beta-strands and the amino-terminal end of the alfa-helices (2). We have additionally shown that the wild-type Chit42 can recognize animal like, beta-1,4-galactosylated and alfa-1,3-fucosylated chito-oligosaccharides (2). Our aim is to investigate the interactions of the T. harzianum Chit42 with natural and modified chito-oligosaccharides using wild-type and different engineered chitinase variants. The work will involve heterologous expression of the chitinase cDNA in E.coli, protein engineering by rational design or by directed evolution methods, and characterising the wild-type and mutant chitinases for their activity and binding specificity. The characterisation of the protein-carbohydrate interactions will include the usage of atomic force microscopy force-distance curves.",
author = "Michael Lienemann and Harry Boer and Arja Paananen and Anu Koivula",
note = "Project : 105-B5SU00412-STI ; 9th European Training Course on Carbohydrates ; Conference date: 06-06-2006 Through 09-06-2006",
year = "2006",
language = "English",

}

Lienemann, M, Boer, H, Paananen, A & Koivula, A 2006, 'Studies on the protein-carbohydrate interaction of the Trichoderma harzianum chitinase Chit42' 9th European Training Course on Carbohydrates, Wageningen, Netherlands, 6/06/06 - 9/06/06, .

Studies on the protein-carbohydrate interaction of the Trichoderma harzianum chitinase Chit42. / Lienemann, Michael; Boer, Harry; Paananen, Arja; Koivula, Anu.

2006. Abstract from 9th European Training Course on Carbohydrates, Wageningen, Netherlands.

Research output: Contribution to conferenceConference AbstractScientific

TY - CONF

T1 - Studies on the protein-carbohydrate interaction of the Trichoderma harzianum chitinase Chit42

AU - Lienemann, Michael

AU - Boer, Harry

AU - Paananen, Arja

AU - Koivula, Anu

N1 - Project : 105-B5SU00412-STI

PY - 2006

Y1 - 2006

N2 - Trichoderma harzianum is a filamentous fungus that can secrete several chitinases (EC 3.2.1.14) to the culture medium under inductive conditions. T. harzianum Chit42 is among the best studied Trichoderma chitinases hydrolysing polymeric chitin mainly into chitobiose, a disaccharide. Chit42 is a single catalytic module enzyme, which belongs to the glycosyl hydrolase family 18 having a (beta/alfa)8 barrel fold. A three-dimensional structural model of T. harzianum Chit42 was built based on the 53% amino acid sequence identity with another fungal chitinase CiX1 from Coccidioides immitis, whose crystal structure has been determined (1). Our modelling studies suggested that Chit42 has at least 7 subsites (-5-4-3-2-1+1+2) for binding the polymeric substrate, and the substrate binding groove is formed by loops positioned between the carboxy-terminal end of the beta-strands and the amino-terminal end of the alfa-helices (2). We have additionally shown that the wild-type Chit42 can recognize animal like, beta-1,4-galactosylated and alfa-1,3-fucosylated chito-oligosaccharides (2). Our aim is to investigate the interactions of the T. harzianum Chit42 with natural and modified chito-oligosaccharides using wild-type and different engineered chitinase variants. The work will involve heterologous expression of the chitinase cDNA in E.coli, protein engineering by rational design or by directed evolution methods, and characterising the wild-type and mutant chitinases for their activity and binding specificity. The characterisation of the protein-carbohydrate interactions will include the usage of atomic force microscopy force-distance curves.

AB - Trichoderma harzianum is a filamentous fungus that can secrete several chitinases (EC 3.2.1.14) to the culture medium under inductive conditions. T. harzianum Chit42 is among the best studied Trichoderma chitinases hydrolysing polymeric chitin mainly into chitobiose, a disaccharide. Chit42 is a single catalytic module enzyme, which belongs to the glycosyl hydrolase family 18 having a (beta/alfa)8 barrel fold. A three-dimensional structural model of T. harzianum Chit42 was built based on the 53% amino acid sequence identity with another fungal chitinase CiX1 from Coccidioides immitis, whose crystal structure has been determined (1). Our modelling studies suggested that Chit42 has at least 7 subsites (-5-4-3-2-1+1+2) for binding the polymeric substrate, and the substrate binding groove is formed by loops positioned between the carboxy-terminal end of the beta-strands and the amino-terminal end of the alfa-helices (2). We have additionally shown that the wild-type Chit42 can recognize animal like, beta-1,4-galactosylated and alfa-1,3-fucosylated chito-oligosaccharides (2). Our aim is to investigate the interactions of the T. harzianum Chit42 with natural and modified chito-oligosaccharides using wild-type and different engineered chitinase variants. The work will involve heterologous expression of the chitinase cDNA in E.coli, protein engineering by rational design or by directed evolution methods, and characterising the wild-type and mutant chitinases for their activity and binding specificity. The characterisation of the protein-carbohydrate interactions will include the usage of atomic force microscopy force-distance curves.

M3 - Conference Abstract

ER -

Lienemann M, Boer H, Paananen A, Koivula A. Studies on the protein-carbohydrate interaction of the Trichoderma harzianum chitinase Chit42. 2006. Abstract from 9th European Training Course on Carbohydrates, Wageningen, Netherlands.