A comparative study of hydrolysis and transglycosylation activities of fungal beta-glucosidases

Christina Helena Bohlin, Peter Westh, Martin Johannes Baumann, Eigil Præstgaard, Kim Borch, Jens Praestgaard, Monrad Rune N

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

β-glucosidases (BGs) from Aspergillus fumigatus, Aspergillus niger, Aspergillus oryzae, Magnaporthe grisea, Neurospora crassa, and Penicillium brasilianum were purified to homogeneity, and investigated for their (simultaneous) hydrolytic and transglycosylation activity in samples with high concentrations of either cellobiose or glucose. The rate of the hydrolytic process (which converts one cellobiose to two glucose molecules) shows a maximum around 10–15 mM cellobiose and decreases with further increase in the concentration of substrate. At the highest investigated concentration (100 mM cellobiose), the hydrolytic activity for the different enzymes ranged from 10% to 55% of the maximum value. This decline in hydrolysis was essentially compensated by increased transglycosylation (which converts two cellobiose to one glucose and one trisaccharide). Hence, it was concluded that the hydrolytic slowdown at high substrate concentrations solely relies on an increased flow through the transglycosylation pathway and not an inhibition that delays the catalytic cycle. Transglycosylation was also detected at high product (glucose) concentrations, but in this case, it was not a major cause for the slowdown in hydrolysis. The experimental data was modeled to obtain kinetic parameters for both hydrolysis and transglycosylation. These parameters were subsequently used in calculations that quantified the negative effects on BG activity of respectively transglycosylation and product inhibition. The kinetic parameters and the mathematical method presented here allow estimation of these effects, and we suggest that this may be useful for the evaluation of BGs for industrial use.
OriginalsprogDansk
TidsskriftApplied Microbiology and Biotechnology
Vol/bind97
Udgave nummer1
Sider (fra-til)159-169
ISSN0175-7598
DOI
StatusUdgivet - jan. 2013

Citer dette

Bohlin, Christina Helena ; Westh, Peter ; Baumann, Martin Johannes ; Præstgaard, Eigil ; Borch, Kim ; Praestgaard, Jens ; Rune N, Monrad. / A comparative study of hydrolysis and transglycosylation activities of fungal beta-glucosidases. I: Applied Microbiology and Biotechnology. 2013 ; Bind 97, Nr. 1. s. 159-169.
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A comparative study of hydrolysis and transglycosylation activities of fungal beta-glucosidases. / Bohlin, Christina Helena; Westh, Peter; Baumann, Martin Johannes; Præstgaard, Eigil; Borch, Kim; Praestgaard, Jens ; Rune N, Monrad.

I: Applied Microbiology and Biotechnology, Bind 97, Nr. 1, 01.2013, s. 159-169.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - A comparative study of hydrolysis and transglycosylation activities of fungal beta-glucosidases

AU - Bohlin, Christina Helena

AU - Westh, Peter

AU - Baumann, Martin Johannes

AU - Præstgaard, Eigil

AU - Borch, Kim

AU - Praestgaard, Jens

AU - Rune N, Monrad

PY - 2013/1

Y1 - 2013/1

N2 - β-glucosidases (BGs) from Aspergillus fumigatus, Aspergillus niger, Aspergillus oryzae, Magnaporthe grisea, Neurospora crassa, and Penicillium brasilianum were purified to homogeneity, and investigated for their (simultaneous) hydrolytic and transglycosylation activity in samples with high concentrations of either cellobiose or glucose. The rate of the hydrolytic process (which converts one cellobiose to two glucose molecules) shows a maximum around 10–15 mM cellobiose and decreases with further increase in the concentration of substrate. At the highest investigated concentration (100 mM cellobiose), the hydrolytic activity for the different enzymes ranged from 10% to 55% of the maximum value. This decline in hydrolysis was essentially compensated by increased transglycosylation (which converts two cellobiose to one glucose and one trisaccharide). Hence, it was concluded that the hydrolytic slowdown at high substrate concentrations solely relies on an increased flow through the transglycosylation pathway and not an inhibition that delays the catalytic cycle. Transglycosylation was also detected at high product (glucose) concentrations, but in this case, it was not a major cause for the slowdown in hydrolysis. The experimental data was modeled to obtain kinetic parameters for both hydrolysis and transglycosylation. These parameters were subsequently used in calculations that quantified the negative effects on BG activity of respectively transglycosylation and product inhibition. The kinetic parameters and the mathematical method presented here allow estimation of these effects, and we suggest that this may be useful for the evaluation of BGs for industrial use.

AB - β-glucosidases (BGs) from Aspergillus fumigatus, Aspergillus niger, Aspergillus oryzae, Magnaporthe grisea, Neurospora crassa, and Penicillium brasilianum were purified to homogeneity, and investigated for their (simultaneous) hydrolytic and transglycosylation activity in samples with high concentrations of either cellobiose or glucose. The rate of the hydrolytic process (which converts one cellobiose to two glucose molecules) shows a maximum around 10–15 mM cellobiose and decreases with further increase in the concentration of substrate. At the highest investigated concentration (100 mM cellobiose), the hydrolytic activity for the different enzymes ranged from 10% to 55% of the maximum value. This decline in hydrolysis was essentially compensated by increased transglycosylation (which converts two cellobiose to one glucose and one trisaccharide). Hence, it was concluded that the hydrolytic slowdown at high substrate concentrations solely relies on an increased flow through the transglycosylation pathway and not an inhibition that delays the catalytic cycle. Transglycosylation was also detected at high product (glucose) concentrations, but in this case, it was not a major cause for the slowdown in hydrolysis. The experimental data was modeled to obtain kinetic parameters for both hydrolysis and transglycosylation. These parameters were subsequently used in calculations that quantified the negative effects on BG activity of respectively transglycosylation and product inhibition. The kinetic parameters and the mathematical method presented here allow estimation of these effects, and we suggest that this may be useful for the evaluation of BGs for industrial use.

U2 - 10.1007/s00253-012-3875-9

DO - 10.1007/s00253-012-3875-9

M3 - Tidsskriftartikel

VL - 97

SP - 159

EP - 169

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

SN - 0175-7598

IS - 1

ER -