PUL ID

PUL0434

PubMed

10613891, J Bacteriol. 2000 Jan;182(1):244-7. doi: 10.1128/jb.182.1.244-247.2000.

Characterization method

SDS-PAGE, Western Blot

Genomic accession number

AF132735.2

Nucelotide position range

91-14184

Substrate

mannooligosaccharide

Loci

engK-hbpA-engL-manA-engM

Species

Clostridium cellulovorans/1493

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

engK - AAF06107.1 91 - 2770 (+) AF132735.2:182-2861 3.2.1.4
hbpA - AAF06108.1 2917 - 3640 (+) AF132735.2:3008-3731 -
engL - AAF06109.1 3873 - 5442 (+) AF132735.2:3964-5533 3.2.1.4
manA - AAF06110.2 6012 - 7290 (+) AF132735.2:6103-7381 3.2.1.78
engM - AAF06111.2 8124 - 10755 (+) AF132735.2:8215-10846 -
engN - AAF61310.1 11111 - 11717 (+) AF132735.2:11202-11808 -
trp - AAF61311.1 12100 - 13042 (-) AF132735.2:12191-13133 -
mln - AAF61312.1 13313 - 14184 (+) AF132735.2:13404-14275 -

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

- 92 - 2770 (+) CAZyme: CBM30|CBM4|GH9 Yes
- 2918 - 3640 (+) other Yes
- 3874 - 5442 (+) CAZyme: GH9 Yes
- 6013 - 7290 (+) CAZyme: GH5_17 Yes
- 8125 - 10755 (+) CAZyme: CBM4|GH9 Yes
- 11112 - 11717 (+) CAZyme: CBM4 Yes
- 12101 - 13042 (-) other Yes
- 13314 - 14184 (+) TC: gnl|TC-DB|Q48797|2.A.69.4.5 Yes

PUL ID

PUL0434

PubMed

10613891, J Bacteriol. 2000 Jan;182(1):244-7. doi: 10.1128/jb.182.1.244-247.2000.

Title

The engL gene cluster of Clostridium cellulovorans contains a gene for cellulosomal manA.

Author

Tamaru Y, Doi RH

Abstract

A five-gene cluster around the gene in Clostridium cellulovorans that encodes endoglucanase EngL, which is involved in plant cell wall degradation, has been cloned and sequenced. As a result, a mannanase gene, manA, has been found downstream of engL. The manA gene consists of an open reading frame with 1,275 nucleotides encoding a protein with 425 amino acids and a molecular weight of 47, 156. ManA has a signal peptide followed by a duplicated sequence (DS, or dockerin) at its N terminus and a catalytic domain which belongs to family 5 of the glycosyl hydrolases and shows high sequence similarity with fungal mannanases, such as Agaricus bisporus Cel4 (17.3% identity), Aspergillus aculeatus Man1 (23.7% identity), and Trichoderma reesei Man1 (22.7% identity). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and N-terminal amino acid sequence analyses of the purified recombinant ManA (rManA) indicated that the N-terminal region of the rManA contained a DS and was truncated in Escherichia coli cells. Furthermore, Western blot analysis indicated that ManA is one of the cellulosomal subunits. ManA production is repressed by cellobiose.