Anomeric Selectivity and Product Profile of a Processive Cellulase

Jeppe Kari, Riin Kont, Kim Borch, Steen Buskov, Johan Pelck Olsen, Nicolaj Cruyz-Bagger, Priit Väljamäe, Peter Westh

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Resumé

Cellobiohydrolases (CBHs) make up an important group of enzymes for both natural carbon cycling and industrial deconstruction of lignocellulosic biomass. The consecutive hydrolysis of one cellulose strand relies on an intricate pattern of enzyme–substrate interactions in the long, tunnel-shaped binding site of the CBH. In this work, we have investigated the initial complexation mode with cellulose of the most thoroughly studied CBH, Cel7A from Hypocrea jecorina (HjCel7A). We found that HjCel7A predominantly produces glucose when it initiates a processive run on insoluble microcrystalline cellulose, confirming the validity of an even and odd product ratio as an estimate of processivity. Moreover, the glucose released from cellulose was predominantly α-glucose. A link between the initial binding mode of the enzyme and the reducing end configuration was investigated by inhibition studies with the two anomers of cellobiose. A clear preference for β-cellobiose in product binding site +2 was observed for HjCel7A, but not the homologous endoglucanase, HjCe7B. Possible relationships between this anomeric preference in the product site and the prevalence of odd-numbered initial-cut products are discussed, and a correlation between processivity and anomer selectivity is proposed.
OriginalsprogEngelsk
TidsskriftBiochemistry
Vol/bind56
Udgave nummer1
Sider (fra-til)167-178
Antal sider11
ISSN0006-2960
DOI
StatusUdgivet - 2017

Citer dette

Kari, Jeppe ; Kont, Riin ; Borch, Kim ; Buskov, Steen ; Olsen, Johan Pelck ; Cruyz-Bagger, Nicolaj ; Väljamäe, Priit ; Westh, Peter. / Anomeric Selectivity and Product Profile of a Processive Cellulase. I: Biochemistry. 2017 ; Bind 56, Nr. 1. s. 167-178.
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abstract = "Cellobiohydrolases (CBHs) make up an important group of enzymes for both natural carbon cycling and industrial deconstruction of lignocellulosic biomass. The consecutive hydrolysis of one cellulose strand relies on an intricate pattern of enzyme–substrate interactions in the long, tunnel-shaped binding site of the CBH. In this work, we have investigated the initial complexation mode with cellulose of the most thoroughly studied CBH, Cel7A from Hypocrea jecorina (HjCel7A). We found that HjCel7A predominantly produces glucose when it initiates a processive run on insoluble microcrystalline cellulose, confirming the validity of an even and odd product ratio as an estimate of processivity. Moreover, the glucose released from cellulose was predominantly α-glucose. A link between the initial binding mode of the enzyme and the reducing end configuration was investigated by inhibition studies with the two anomers of cellobiose. A clear preference for β-cellobiose in product binding site +2 was observed for HjCel7A, but not the homologous endoglucanase, HjCe7B. Possible relationships between this anomeric preference in the product site and the prevalence of odd-numbered initial-cut products are discussed, and a correlation between processivity and anomer selectivity is proposed.",
author = "Jeppe Kari and Riin Kont and Kim Borch and Steen Buskov and Olsen, {Johan Pelck} and Nicolaj Cruyz-Bagger and Priit V{\"a}ljam{\"a}e and Peter Westh",
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Kari, J, Kont, R, Borch, K, Buskov, S, Olsen, JP, Cruyz-Bagger, N, Väljamäe, P & Westh, P 2017, 'Anomeric Selectivity and Product Profile of a Processive Cellulase', Biochemistry, bind 56, nr. 1, s. 167-178. https://doi.org/10.1021/acs.biochem.6b00636

Anomeric Selectivity and Product Profile of a Processive Cellulase. / Kari, Jeppe; Kont, Riin; Borch, Kim; Buskov, Steen; Olsen, Johan Pelck; Cruyz-Bagger, Nicolaj; Väljamäe, Priit; Westh, Peter.

I: Biochemistry, Bind 56, Nr. 1, 2017, s. 167-178.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Anomeric Selectivity and Product Profile of a Processive Cellulase

AU - Kari, Jeppe

AU - Kont, Riin

AU - Borch, Kim

AU - Buskov, Steen

AU - Olsen, Johan Pelck

AU - Cruyz-Bagger, Nicolaj

AU - Väljamäe, Priit

AU - Westh, Peter

N1 - PMID: 28026938

PY - 2017

Y1 - 2017

N2 - Cellobiohydrolases (CBHs) make up an important group of enzymes for both natural carbon cycling and industrial deconstruction of lignocellulosic biomass. The consecutive hydrolysis of one cellulose strand relies on an intricate pattern of enzyme–substrate interactions in the long, tunnel-shaped binding site of the CBH. In this work, we have investigated the initial complexation mode with cellulose of the most thoroughly studied CBH, Cel7A from Hypocrea jecorina (HjCel7A). We found that HjCel7A predominantly produces glucose when it initiates a processive run on insoluble microcrystalline cellulose, confirming the validity of an even and odd product ratio as an estimate of processivity. Moreover, the glucose released from cellulose was predominantly α-glucose. A link between the initial binding mode of the enzyme and the reducing end configuration was investigated by inhibition studies with the two anomers of cellobiose. A clear preference for β-cellobiose in product binding site +2 was observed for HjCel7A, but not the homologous endoglucanase, HjCe7B. Possible relationships between this anomeric preference in the product site and the prevalence of odd-numbered initial-cut products are discussed, and a correlation between processivity and anomer selectivity is proposed.

AB - Cellobiohydrolases (CBHs) make up an important group of enzymes for both natural carbon cycling and industrial deconstruction of lignocellulosic biomass. The consecutive hydrolysis of one cellulose strand relies on an intricate pattern of enzyme–substrate interactions in the long, tunnel-shaped binding site of the CBH. In this work, we have investigated the initial complexation mode with cellulose of the most thoroughly studied CBH, Cel7A from Hypocrea jecorina (HjCel7A). We found that HjCel7A predominantly produces glucose when it initiates a processive run on insoluble microcrystalline cellulose, confirming the validity of an even and odd product ratio as an estimate of processivity. Moreover, the glucose released from cellulose was predominantly α-glucose. A link between the initial binding mode of the enzyme and the reducing end configuration was investigated by inhibition studies with the two anomers of cellobiose. A clear preference for β-cellobiose in product binding site +2 was observed for HjCel7A, but not the homologous endoglucanase, HjCe7B. Possible relationships between this anomeric preference in the product site and the prevalence of odd-numbered initial-cut products are discussed, and a correlation between processivity and anomer selectivity is proposed.

U2 - 10.1021/acs.biochem.6b00636

DO - 10.1021/acs.biochem.6b00636

M3 - Journal article

VL - 56

SP - 167

EP - 178

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 1

ER -

Kari J, Kont R, Borch K, Buskov S, Olsen JP, Cruyz-Bagger N et al. Anomeric Selectivity and Product Profile of a Processive Cellulase. Biochemistry. 2017;56(1):167-178. https://doi.org/10.1021/acs.biochem.6b00636