TY - JOUR
T1 - Cellodextrin phosphorylase from Ruminiclostridium thermocellum
T2 - X-ray crystal structure and substrate specificity analysis
AU - O'Neill, Ellis C.
AU - Pergolizzi, Giulia
AU - Stevenson, Clare E.M.
AU - Lawson, David M.
AU - Nepogodiev, Sergey A.
AU - Field, Robert A.
N1 - Funding Information:
These studies were supported by the UK BBSRC Institute Strategic Program on Understanding and Exploiting Metabolism (MET) [BB/J004561/1] and the John Innes Foundation; the BBSRC, EPSRC and InnovateUK: IBCatalyst [BB/M02903411]. We acknowledge the Diamond Light Source for access to beamline I02 under proposal MX7641.
Funding Information:
These studies were supported by the UK BBSRC Institute Strategic Program on Understanding and Exploiting Metabolism (MET) [ BB/J004561/1 ] and the John Innes Foundation ; the BBSRC, EPSRC and InnovateUK: IBCatalyst [ BB/M02903411 ]. We acknowledge the Diamond Light Source for access to beamline I02 under proposal MX7641.
Publisher Copyright:
© 2017 The Authors
PY - 2017/11/8
Y1 - 2017/11/8
N2 - The GH94 glycoside hydrolase cellodextrin phosphorylase (CDP, EC 2.4.1.49) produces cellodextrin oligomers from short β-1→4-glucans and α-D-glucose 1-phosphate. Compared to cellobiose phosphorylase (CBP), which produces cellobiose from glucose and α-D-glucose 1-phosphate, CDP is biochemically less well characterised. Herein, we investigate the donor and acceptor substrate specificity of recombinant CDP from Ruminiclostridium thermocellum and we isolate and characterise a glucosamine addition product to the cellobiose acceptor with the non-natural donor α-D-glucosamine 1-phosphate. In addition, we report the first X-ray crystal structure of CDP, along with comparison to the available structures from CBPs and other closely related enzymes, which contributes to understanding of the key structural features necessary to discriminate between monosaccharide (CBP) and oligosaccharide (CDP) acceptor substrates.
AB - The GH94 glycoside hydrolase cellodextrin phosphorylase (CDP, EC 2.4.1.49) produces cellodextrin oligomers from short β-1→4-glucans and α-D-glucose 1-phosphate. Compared to cellobiose phosphorylase (CBP), which produces cellobiose from glucose and α-D-glucose 1-phosphate, CDP is biochemically less well characterised. Herein, we investigate the donor and acceptor substrate specificity of recombinant CDP from Ruminiclostridium thermocellum and we isolate and characterise a glucosamine addition product to the cellobiose acceptor with the non-natural donor α-D-glucosamine 1-phosphate. In addition, we report the first X-ray crystal structure of CDP, along with comparison to the available structures from CBPs and other closely related enzymes, which contributes to understanding of the key structural features necessary to discriminate between monosaccharide (CBP) and oligosaccharide (CDP) acceptor substrates.
KW - Cellodextrin phosphorylase
KW - Glucosamine 1-phosphate
KW - X-ray crystal structure
UR - http://www.scopus.com/inward/record.url?scp=85026300425&partnerID=8YFLogxK
U2 - 10.1016/j.carres.2017.07.005
DO - 10.1016/j.carres.2017.07.005
M3 - Article
C2 - 28760417
AN - SCOPUS:85026300425
VL - 451
SP - 118
EP - 132
JO - Carbohydrate Research
JF - Carbohydrate Research
SN - 0008-6215
ER -