Effects of pH and high ionic strength on the adsorption and activity of native and mutated cellobiohydrolase I from Trichoderma reesei

Tapani Reinikainen (Corresponding Author), Olle Teleman, Tuula Teeri

Research output: Contribution to journalArticleScientificpeer-review

137 Citations (Scopus)

Abstract

Cellobiohydrolase I (CBHI) is the major cellulase of Trichoderma reesei. The enzyme contains a discrete cellulose‐binding domain (CBD), which increases its binding and activity on crystalline cellulose. We studied cellulase‐cellulose interactions using site‐directed mutagenesis on the basis of the three‐dimensional structure of the CBD of CBHI.
Three mutant proteins which have earlier been produced in Saccharomyces cerevisiae were expressed in the native host organism. The data presented here support the hypothesis that a conserved tyrosine (Y492) located on the flat and more hydrophilic surface of the CBD is essential for the functionality.
The data also suggest that the more hydrophobic surface is not directly involved in the CBD function. The pH dependence of the adsorption revealed that electrostatic repulsion between the bound proteins may also control the adsorption.
The binding of CBHI to cellulose was significantly affected by high ionic strength suggesting that the interaction with cellulose includes a hydrophobic effect. High ionic strength increased the activity of the isolated core and of mutant proteins on crystalline cellulose, indicating that once productively bound, the enzymes are capable of solubilizing cellulose even with a mutagenized or with no CBD
Original languageEnglish
Pages (from-to)392-403
JournalProteins
Volume22
Issue number4
DOIs
Publication statusPublished - 1995
MoE publication typeA1 Journal article-refereed

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Cellulose 1,4-beta-Cellobiosidase
Trichoderma
Ionic strength
Cellulose
Osmolar Concentration
Adsorption
Mutant Proteins
Crystalline materials
Mutagenesis
Cellulase
Enzymes
Static Electricity
Yeast
Saccharomyces cerevisiae
Tyrosine
Electrostatics
Proteins

Cite this

Reinikainen, Tapani ; Teleman, Olle ; Teeri, Tuula. / Effects of pH and high ionic strength on the adsorption and activity of native and mutated cellobiohydrolase I from Trichoderma reesei. In: Proteins. 1995 ; Vol. 22, No. 4. pp. 392-403.
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title = "Effects of pH and high ionic strength on the adsorption and activity of native and mutated cellobiohydrolase I from Trichoderma reesei",
abstract = "Cellobiohydrolase I (CBHI) is the major cellulase of Trichoderma reesei. The enzyme contains a discrete cellulose‐binding domain (CBD), which increases its binding and activity on crystalline cellulose. We studied cellulase‐cellulose interactions using site‐directed mutagenesis on the basis of the three‐dimensional structure of the CBD of CBHI. Three mutant proteins which have earlier been produced in Saccharomyces cerevisiae were expressed in the native host organism. The data presented here support the hypothesis that a conserved tyrosine (Y492) located on the flat and more hydrophilic surface of the CBD is essential for the functionality. The data also suggest that the more hydrophobic surface is not directly involved in the CBD function. The pH dependence of the adsorption revealed that electrostatic repulsion between the bound proteins may also control the adsorption. The binding of CBHI to cellulose was significantly affected by high ionic strength suggesting that the interaction with cellulose includes a hydrophobic effect. High ionic strength increased the activity of the isolated core and of mutant proteins on crystalline cellulose, indicating that once productively bound, the enzymes are capable of solubilizing cellulose even with a mutagenized or with no CBD",
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Effects of pH and high ionic strength on the adsorption and activity of native and mutated cellobiohydrolase I from Trichoderma reesei. / Reinikainen, Tapani (Corresponding Author); Teleman, Olle; Teeri, Tuula.

In: Proteins, Vol. 22, No. 4, 1995, p. 392-403.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Effects of pH and high ionic strength on the adsorption and activity of native and mutated cellobiohydrolase I from Trichoderma reesei

AU - Reinikainen, Tapani

AU - Teleman, Olle

AU - Teeri, Tuula

N1 - Project code: BEL4319

PY - 1995

Y1 - 1995

N2 - Cellobiohydrolase I (CBHI) is the major cellulase of Trichoderma reesei. The enzyme contains a discrete cellulose‐binding domain (CBD), which increases its binding and activity on crystalline cellulose. We studied cellulase‐cellulose interactions using site‐directed mutagenesis on the basis of the three‐dimensional structure of the CBD of CBHI. Three mutant proteins which have earlier been produced in Saccharomyces cerevisiae were expressed in the native host organism. The data presented here support the hypothesis that a conserved tyrosine (Y492) located on the flat and more hydrophilic surface of the CBD is essential for the functionality. The data also suggest that the more hydrophobic surface is not directly involved in the CBD function. The pH dependence of the adsorption revealed that electrostatic repulsion between the bound proteins may also control the adsorption. The binding of CBHI to cellulose was significantly affected by high ionic strength suggesting that the interaction with cellulose includes a hydrophobic effect. High ionic strength increased the activity of the isolated core and of mutant proteins on crystalline cellulose, indicating that once productively bound, the enzymes are capable of solubilizing cellulose even with a mutagenized or with no CBD

AB - Cellobiohydrolase I (CBHI) is the major cellulase of Trichoderma reesei. The enzyme contains a discrete cellulose‐binding domain (CBD), which increases its binding and activity on crystalline cellulose. We studied cellulase‐cellulose interactions using site‐directed mutagenesis on the basis of the three‐dimensional structure of the CBD of CBHI. Three mutant proteins which have earlier been produced in Saccharomyces cerevisiae were expressed in the native host organism. The data presented here support the hypothesis that a conserved tyrosine (Y492) located on the flat and more hydrophilic surface of the CBD is essential for the functionality. The data also suggest that the more hydrophobic surface is not directly involved in the CBD function. The pH dependence of the adsorption revealed that electrostatic repulsion between the bound proteins may also control the adsorption. The binding of CBHI to cellulose was significantly affected by high ionic strength suggesting that the interaction with cellulose includes a hydrophobic effect. High ionic strength increased the activity of the isolated core and of mutant proteins on crystalline cellulose, indicating that once productively bound, the enzymes are capable of solubilizing cellulose even with a mutagenized or with no CBD

U2 - 10.1002/prot.340220409

DO - 10.1002/prot.340220409

M3 - Article

VL - 22

SP - 392

EP - 403

JO - Proteins

JF - Proteins

SN - 0887-3585

IS - 4

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