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Cite: NAR/gkaa742 2020

Because CGCFinder predicted no CGC for this PUL, the gene cluster depicted below contains dbCAN2 and CGC signature predictions for all genes in the PUL, instead of a predicted CGC.


PUL General Information

Literature Information

PUL ID

PUL0011

PubMed

8936327, Microbiology (Reading). 1996 Apr;142 ( Pt 4):1013-1023. doi: 10.1099/00221287-142-4-1013.

Characterization method

enzyme activity assay

Genomic accession number

NC_011898.1

Nucelotide position range

843032-851723

Substrate

cellulose

Loci

CCEL_RS03700-CCEL_RS03715

Species

Ruminiclostridium cellulolyticum/1521

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

- CCEL_RS03700 WP_015924275.1 0 - 2169 (+) NC_011898.1:843032-845201 -
- CCEL_RS03705 WP_015924276.1 2297 - 3680 (+) NC_011898.1:845329-846712 -
- CCEL_RS03710 WP_015924277.1 3764 - 5942 (+) NC_011898.1:846796-848974 -
- CCEL_RS03715 WP_015924278.1 6034 - 8692 (+) NC_011898.1:849066-851724 -

Cluster number

0

Gene name

Gene position

Gene type

Found by CGCFinder?

- 1 - 2169 (+) CAZyme: GH48 No
- 2298 - 3680 (+) CAZyme: GH8 No
- 3765 - 5942 (+) CAZyme: CBM3|GH9 No
- 6035 - 8692 (+) CAZyme: CBM4|GH9 No

PUL ID

PUL0011

PubMed

8936327, Microbiology (Reading). 1996 Apr;142 ( Pt 4):1013-1023. doi: 10.1099/00221287-142-4-1013.

Title

Molecular study and overexpression of the Clostridium cellulolyticum celF cellulase gene in Escherichia coli.

Author

Reverbel-Leroy C, Belaich A, Bernadac A, Gaudin C, Belaich JP, Tardif C

Abstract

The CelF-encoding sequence was isolated from Clostridium cellulolyticum genomic DNA using the inverse PCR technique. The gene lies between cipC (the gene encoding the cellulosome scaffolding protein) and celC (coding for the endoglucanase C) in the large cel cluster of this mesophilic cellulolytic Clostridium species. Comparisons between the deduced amino acid sequence of the mature CelF (693 amino acids, molecular mass 77626) and those of other beta-glycanases showed that this enzyme belongs to the recently proposed family L of cellulases (family 48 of glycosyl hydrolases). The protein was overproduced in Escherichia coli using the T7 expression system. It formed both cytoplasmic and periplasmic inclusion bodies when induction was performed at 37 degrees C. Surprisingly, the protein synthesized from the cytoplasmic production vector was degraded in the Ion protease-deficient strain BL21(DE3). The induction conditions were optimized with regard to the concentration of inductor, cell density, and temperature and time of induction in order to overproduce an active periplasmic protein (CelFp) which was both soluble and stable. It was collected using the osmotic shock method. The enzymic degradation of various cellulosic substrates by CelFp was studied. CelFp degraded swollen Avicel more efficiently than substituted soluble CM-cellulose or crystalline Avicel and was not active on xylan. Its activity is therefore quite different from that of endoglucanases, which are most active on CM-cellulose.