Inhibition of enzymatic hydrolysis by residual lignins from softwood: Study of enzyme binding and inactivation on lignin-rich surface

Jenni Rahikainen, Saara Mikander, Kaisa Marjamaa, Tarja Tamminen, Angelos Lappas, Liisa Viikari, Kristiina Kruus (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

173 Citations (Scopus)

Abstract

Lignin‐derived inhibition is a major obstacle restricting the enzymatic hydrolysis of cell wall polysaccharides especially with softwood lignocellulosics. Enzyme adsorption on lignin is suggested to contribute to the inhibitory effect of lignin. The interaction of cellulases with softwood lignin was studied in the present work with commercial Trichoderma reesei cellulases (Celluclast) and lignin‐rich residues isolated from steam pretreated softwood (SPS) by enzymatic and acid hydrolysis. Both lignin preparations inhibited the hydrolysis of microcrystalline cellulose (Avicel) and adsorbed the major cellulases present in the commercial cellulase mixture. The adsorption phenomenon was studied at low temperature (4°C) and at the typical hydrolysis temperature (45°C) by following activities of free and lignin‐bound enzymes. Severe inactivation of the lignin‐bound enzymes was observed at 45°C, however at 4°C the enzymes retained well their activity. Furthermore, SDS–PAGE analysis of the lignin‐bound enzymes indicated that very strong interactions form between the residue and the enzymes at 45°C, because the enzymes were not released from the residue in the electrophoresis. These results suggest that heat‐induced denaturation may take place on the surface of softwood lignin at the hydrolysis temperature.
Original languageEnglish
Pages (from-to)2823-2834
Number of pages12
JournalBiotechnology and Bioengineering
Volume108
Issue number12
DOIs
Publication statusPublished - 2011
MoE publication typeA1 Journal article-refereed

Fingerprint

Enzyme inhibition
Enzymatic hydrolysis
Softwoods
Lignin
Hydrolysis
Enzymes
Cellulases
Adsorption
Temperature
Trichoderma
Denaturation
Cellulase
Steam
Polysaccharides
Electrophoresis
Cellulose
Cell Wall
Catalyst activity
Cells
Acids

Keywords

  • lignocellulose
  • softwood lignin
  • enzymatic hydrolysis
  • cellulase
  • cellulase adsorption
  • Trichoderma reesei
  • denaturation

Cite this

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title = "Inhibition of enzymatic hydrolysis by residual lignins from softwood: Study of enzyme binding and inactivation on lignin-rich surface",
abstract = "Lignin‐derived inhibition is a major obstacle restricting the enzymatic hydrolysis of cell wall polysaccharides especially with softwood lignocellulosics. Enzyme adsorption on lignin is suggested to contribute to the inhibitory effect of lignin. The interaction of cellulases with softwood lignin was studied in the present work with commercial Trichoderma reesei cellulases (Celluclast) and lignin‐rich residues isolated from steam pretreated softwood (SPS) by enzymatic and acid hydrolysis. Both lignin preparations inhibited the hydrolysis of microcrystalline cellulose (Avicel) and adsorbed the major cellulases present in the commercial cellulase mixture. The adsorption phenomenon was studied at low temperature (4°C) and at the typical hydrolysis temperature (45°C) by following activities of free and lignin‐bound enzymes. Severe inactivation of the lignin‐bound enzymes was observed at 45°C, however at 4°C the enzymes retained well their activity. Furthermore, SDS–PAGE analysis of the lignin‐bound enzymes indicated that very strong interactions form between the residue and the enzymes at 45°C, because the enzymes were not released from the residue in the electrophoresis. These results suggest that heat‐induced denaturation may take place on the surface of softwood lignin at the hydrolysis temperature.",
keywords = "lignocellulose, softwood lignin, enzymatic hydrolysis, cellulase, cellulase adsorption, Trichoderma reesei, denaturation",
author = "Jenni Rahikainen and Saara Mikander and Kaisa Marjamaa and Tarja Tamminen and Angelos Lappas and Liisa Viikari and Kristiina Kruus",
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Inhibition of enzymatic hydrolysis by residual lignins from softwood : Study of enzyme binding and inactivation on lignin-rich surface. / Rahikainen, Jenni; Mikander, Saara; Marjamaa, Kaisa; Tamminen, Tarja; Lappas, Angelos; Viikari, Liisa; Kruus, Kristiina (Corresponding Author).

In: Biotechnology and Bioengineering, Vol. 108, No. 12, 2011, p. 2823-2834.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Inhibition of enzymatic hydrolysis by residual lignins from softwood

T2 - Study of enzyme binding and inactivation on lignin-rich surface

AU - Rahikainen, Jenni

AU - Mikander, Saara

AU - Marjamaa, Kaisa

AU - Tamminen, Tarja

AU - Lappas, Angelos

AU - Viikari, Liisa

AU - Kruus, Kristiina

PY - 2011

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N2 - Lignin‐derived inhibition is a major obstacle restricting the enzymatic hydrolysis of cell wall polysaccharides especially with softwood lignocellulosics. Enzyme adsorption on lignin is suggested to contribute to the inhibitory effect of lignin. The interaction of cellulases with softwood lignin was studied in the present work with commercial Trichoderma reesei cellulases (Celluclast) and lignin‐rich residues isolated from steam pretreated softwood (SPS) by enzymatic and acid hydrolysis. Both lignin preparations inhibited the hydrolysis of microcrystalline cellulose (Avicel) and adsorbed the major cellulases present in the commercial cellulase mixture. The adsorption phenomenon was studied at low temperature (4°C) and at the typical hydrolysis temperature (45°C) by following activities of free and lignin‐bound enzymes. Severe inactivation of the lignin‐bound enzymes was observed at 45°C, however at 4°C the enzymes retained well their activity. Furthermore, SDS–PAGE analysis of the lignin‐bound enzymes indicated that very strong interactions form between the residue and the enzymes at 45°C, because the enzymes were not released from the residue in the electrophoresis. These results suggest that heat‐induced denaturation may take place on the surface of softwood lignin at the hydrolysis temperature.

AB - Lignin‐derived inhibition is a major obstacle restricting the enzymatic hydrolysis of cell wall polysaccharides especially with softwood lignocellulosics. Enzyme adsorption on lignin is suggested to contribute to the inhibitory effect of lignin. The interaction of cellulases with softwood lignin was studied in the present work with commercial Trichoderma reesei cellulases (Celluclast) and lignin‐rich residues isolated from steam pretreated softwood (SPS) by enzymatic and acid hydrolysis. Both lignin preparations inhibited the hydrolysis of microcrystalline cellulose (Avicel) and adsorbed the major cellulases present in the commercial cellulase mixture. The adsorption phenomenon was studied at low temperature (4°C) and at the typical hydrolysis temperature (45°C) by following activities of free and lignin‐bound enzymes. Severe inactivation of the lignin‐bound enzymes was observed at 45°C, however at 4°C the enzymes retained well their activity. Furthermore, SDS–PAGE analysis of the lignin‐bound enzymes indicated that very strong interactions form between the residue and the enzymes at 45°C, because the enzymes were not released from the residue in the electrophoresis. These results suggest that heat‐induced denaturation may take place on the surface of softwood lignin at the hydrolysis temperature.

KW - lignocellulose

KW - softwood lignin

KW - enzymatic hydrolysis

KW - cellulase

KW - cellulase adsorption

KW - Trichoderma reesei

KW - denaturation

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JO - Biotechnology and Bioengineering

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SN - 0006-3592

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