Cellobiohydrolase 1 from Trichoderma reesei degrades cellulose in single cellobiose steps.

Abstract:

:Cellobiohydrolase 1 from Trichoderma reesei (TrCel7A) processively hydrolyses cellulose into cellobiose. Although enzymatic techniques have been established as promising tools in biofuel production, a clear understanding of the motor's mechanistic action has yet to be revealed. Here, we develop an optical tweezers-based single-molecule (SM) motility assay for precision tracking of TrCel7A. Direct observation of motility during degradation reveals processive runs and distinct steps on the scale of 1 nm. Our studies suggest TrCel7A is not mechanically limited, can work against 20 pN loads and speeds up when assisted. Temperature-dependent kinetic studies establish the energy requirements for the fundamental stepping cycle, which likely includes energy from glycosidic bonds and other sources. Through SM measurements of isolated TrCel7A domains, we determine that the catalytic domain alone is sufficient for processive motion, providing insight into TrCel7A's molecular motility mechanism.

journal_name

Nat Commun

journal_title

Nature communications

authors

Brady SK,Sreelatha S,Feng Y,Chundawat SP,Lang MJ

doi

10.1038/ncomms10149

subject

Has Abstract

pub_date

2015-12-10 00:00:00

pages

10149

issn

2041-1723

pii

ncomms10149

journal_volume

6

pub_type

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