PUL ID

PUL0445

PubMed

10913091, J Bacteriol. 2000 Aug;182(16):4572-7. doi: 10.1128/jb.182.16.4572-4577.2000.

Characterization method

expression of recombinant proteins, thin layer chromatography, enzyme activity assay

Genomic accession number

AB011415.1

Nucelotide position range

2533-14738

Substrate

alginate

Loci

BAB03312.1-BAB03319.1

Species

Sphingomonas sp./28214

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

aly - BAB03312.1 0 - 1926 (-) AB011415.1:2533-4459 -
ccpA - BAB03313.1 1967 - 3029 (-) AB011415.1:4500-5562 -
algS - BAB03314.1 3231 - 4323 (+) AB011415.1:5764-6856 -
algM1 - BAB03315.1 4417 - 5392 (+) AB011415.1:6950-7925 -
algM2 - BAB03316.1 5448 - 6330 (+) AB011415.1:7981-8863 -
algQ1 - BAB03317.1 6408 - 7989 (+) AB011415.1:8941-10522 -
algQ2 - BAB03318.1 8135 - 9686 (+) AB011415.1:10668-12219 -
oal - BAB03319.1 9920 - 12206 (+) AB011415.1:12453-14739 -

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

- 1 - 1926 (-) CAZyme: PL7|PL5 Yes
- 1968 - 3029 (-) TF: DBD-Pfam|LacI,DBD-SUPERFAMILY|0036955 Yes
- 3232 - 4323 (+) TC: gnl|TC-DB|Q9KWT9|3.A.1.1.10 Yes
- 4418 - 5392 (+) TC: gnl|TC-DB|Q9KWT8|3.A.1.1.10 Yes
- 5449 - 6330 (+) TC: gnl|TC-DB|Q9KWT7|3.A.1.1.10 Yes
- 6409 - 7989 (+) TC: gnl|TC-DB|Q9KWT6|3.A.1.1.10 Yes
- 8136 - 9686 (+) TC: gnl|TC-DB|Q9KWT5|3.A.1.1.10 Yes
- 9921 - 12206 (+) CAZyme: PL15_1 Yes

PUL ID

PUL0445

PubMed

10913091, J Bacteriol. 2000 Aug;182(16):4572-7. doi: 10.1128/jb.182.16.4572-4577.2000.

Title

Molecular identification of oligoalginate lyase of Sphingomonas sp. strain A1 as one of the enzymes required for complete depolymerization of alginate.

Author

Hashimoto W, Miyake O, Momma K, Kawai S, Murata K

Abstract

A bacterium, Sphingomonas sp. strain A1, can incorporate alginate into cells through a novel ABC (ATP-binding cassette) transporter system specific to the macromolecule. The transported alginate is depolymerized to di- and trisaccharides by three kinds of cytoplasmic alginate lyases (A1-I [66 kDa], A1-II [25 kDa], and A1-III [40 kDa]) generated from a single precursor through posttranslational autoprocessing. The resultant alginate oligosaccharides were degraded to monosaccharides by cytoplasmic oligoalginate lyase. The enzyme and its gene were isolated from the bacterial cells grown in the presence of alginate. The purified enzyme was a monomer with a molecular mass of 85 kDa and cleaved glycosidic bonds not only in oligosaccharides produced from alginate by alginate lyases but also in polysaccharides (alginate, polymannuronate, and polyguluronate) most efficiently at pH 8.0 and 37 degrees C. The reaction catalyzed by the oligoalginate lyase was exolytic and thought to play an important role in the complete depolymerization of alginate in Sphingomonas sp. strain A1. The gene for this novel enzyme consisted of an open reading frame of 2,286 bp encoding a polypeptide with a molecular weight of 86,543 and was located downstream of the genes coding for the precursor of alginate lyases (aly) and the ABC transporter (algS, algM1, and algM2). This result indicates that the genes for proteins required for the transport and complete depolymerization of alginate are assembled to form a cluster.