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Title: Orpinomyces cellulase celf protein and coding sequences

Patent ·
OSTI ID:873216
 [1];  [1];  [1]
  1. Athens, GA
+ Show Author Affiliations

A cDNA (1,520 bp), designated celF, consisting of an open reading frame (ORF) encoding a polypeptide (CelF) of 432 amino acids was isolated from a cDNA library of the anaerobic rumen fungus Orpinomyces PC-2 constructed in Escherichia coli. Analysis of the deduced amino acid sequence showed that starting from the N-terminus, CelF consists of a signal peptide, a cellulose binding domain (CBD) followed by an extremely Asn-rich linker region which separate the CBD and the catalytic domains. The latter is located at the C-terminus. The catalytic domain of CelF is highly homologous to CelA and CelC of Orpinomyces PC-2, to CelA of Neocallimastix patriciarum and also to cellobiohydrolase IIs (CBHIIs) from aerobic fungi. However, Like CelA of Neocallimastix patriciarum, CelF does not have the noncatalytic repeated peptide domain (NCRPD) found in CelA and CelC from the same organism. The recombinant protein CelF hydrolyzes cellooligosaccharides in the pattern of CBHII, yielding only cellobiose as product with cellotetraose as the substrate. The genomic celF is interrupted by a 111 bp intron, located within the region coding for the CBD. The intron of the celF has features in common with genes from aerobic filamentous fungi.

Research Organization:
Univ. of Georgia, Athens, GA (United States)
DOE Contract Number:
FG05-93ER20127
Assignee:
University of Georgia Research Foundation, Inc. (Athens, GA)
Patent Number(s):
US 6114158
Application Number:
09/118319
OSTI ID:
873216
Country of Publication:
United States
Language:
English

References (25)

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Related Subjects

orpinomyces
cellulase
celf
protein
coding
sequences
cdna
520
bp
designated
consisting
reading
frame
orf
encoding
polypeptide
432
amino
acids
isolated
library
anaerobic
rumen
fungus
pc-2
constructed
escherichia
coli
analysis
deduced
acid
sequence
starting
n-terminus
consists
signal
peptide
cellulose
binding
domain
cbd
followed
extremely
asn-rich
linker
region
separate
catalytic
domains
latter
located
c-terminus
highly
homologous
cela
celc
neocallimastix
patriciarum
cellobiohydrolase
iis
cbhiis
aerobic
fungi
noncatalytic
repeated
ncrpd
found
organism
recombinant
hydrolyzes
cellooligosaccharides
pattern
cbhii
yielding
cellobiose
product
cellotetraose
substrate
genomic
interrupted
111
intron
features
common
genes
filamentous
deduced amino
reading frame
coding sequences
binding domain
cellulose binding
acid sequence
amino acid
amino acids
cdna library
orpinomyces pc-2
coding sequence
escherichia coli
catalytic domain
orpinomyces cellulase
highly homologous
recombinant protein
/435/999/