In vitro expression of Penicillium janthinellum cellobiohydrolase I gene in a coupled transcription-translation system.

Abstract:

:An enzymatically-active fungal cellobiohydrolase I (CBH I) was first synthesized in a coupled reticulocyte lysate system lacking of glycosylation modification by the template DNA(Cbh1) in the presence of T7 RNA polymerase. The synthesized CBH I had the expected size (57 kDa) and catalyzed the substrate of p-nitrophenyl-beta-D-cellobioside (pNPC), and had no activity against carboxymethyl cellulose (CMC-Na). The K(m) and V(max) values of the CBH I for pNPC were 0.82 mmol and 0.067 micromol min(-1) per microg enzyme, respectively. The results indicated that glycosylation may not be necessary for enzymatic activity of fungal cellulases.

journal_name

J Biotechnol

journal_title

Journal of biotechnology

authors

Wang J,Gao P

doi

10.1016/s0168-1656(00)00285-6

subject

Has Abstract

pub_date

2000-08-25 00:00:00

pages

205-9

issue

2-3

eissn

0168-1656

issn

1873-4863

pii

S0168165600002856

journal_volume

81

pub_type

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