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Cite: NAR/gkaa742 2020



PUL General Information

Literature Information

PUL ID

PUL0196

PubMed

26546429, Appl Environ Microbiol. 2015 Nov 6;82(2):570-7. doi: 10.1128/AEM.02888-15. Print 2016 Jan 15.

Characterization method

enzyme activity assay

Genomic accession number

NC_010999.1

Nucelotide position range

288852-300044

Substrate

lacto-n-triose

Loci

LCABL_RS01420-LCABL_RS01465

Species

Lactobacillus casei/1582

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

- LCABL_RS01420 WP_012490882.1 0 - 1710 (+) NC_010999.1:288852-290562 -
manA LCABL_RS01425 WP_003592893.1 2090 - 3110 (+) NC_010999.1:290942-291962 5.3.1.8
- LCABL_RS01430 WP_012490884.1 3471 - 4188 (+) NC_010999.1:292323-293040 -
- LCABL_RS01435 WP_003563130.1 4256 - 5438 (+) NC_010999.1:293108-294290 -
nagA LCABL_RS01440 WP_003573361.1 5496 - 6657 (+) NC_010999.1:294348-295509 3.5.1.25
- LCABL_RS01445 WP_012490885.1 6756 - 8553 (+) NC_010999.1:295608-297405 -
- LCABL_RS01450 WP_003563136.1 8555 - 9038 (+) NC_010999.1:297407-297890 -
- LCABL_RS01455 WP_003573365.1 9050 - 9968 (+) NC_010999.1:297902-298820 -
- LCABL_RS01460 WP_003563140.1 9954 - 10776 (+) NC_010999.1:298806-299628 -
- LCABL_RS01465 WP_003563142.1 10812 - 11193 (+) NC_010999.1:299664-300045 -

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

- 1 - 1710 (+) CAZyme: GH20 Yes
manA 2091 - 3110 (+) other Yes
- 3472 - 4188 (+) TF: DBD-Pfam|GntR,DBD-SUPERFAMILY|0043758 Yes
- 4257 - 5438 (+) STP: STP|SIS,STP|SIS Yes
nagA 5497 - 6657 (+) CAZyme: CE9 Yes
- 6757 - 8553 (+) TC: gnl|TC-DB|W5L187|9.B.207.2.1 Yes
- 8556 - 9038 (+) TC: gnl|TC-DB|B3W754|4.A.6.1.18 Yes
- 9051 - 9968 (+) TC: gnl|TC-DB|B3W755|4.A.6.1.18 Yes
- 9955 - 10776 (+) TC: gnl|TC-DB|B3W756|4.A.6.1.18 Yes
- 10813 - 11193 (+) TC: gnl|TC-DB|B3W757|4.A.6.1.18 Yes

PUL ID

PUL0196

PubMed

26546429, Appl Environ Microbiol. 2015 Nov 6;82(2):570-7. doi: 10.1128/AEM.02888-15. Print 2016 Jan 15.

Title

The Extracellular Wall-Bound beta-N-Acetylglucosaminidase from Lactobacillus casei Is Involved in the Metabolism of the Human Milk Oligosaccharide Lacto-N-Triose.

Author

Bidart GN, Rodriguez-Diaz J, Yebra MJ

Abstract

Human milk oligosaccharides (HMOs) are considered to play a key role in establishing and maintaining the infant gut microbiota. Lacto-N-triose forms part of both type 1 and type 2 HMOs and also of the glycan moieties of glycoproteins. Upstream of the previously characterized gene cluster involved in lacto-N-biose and galacto-N-biose metabolism from Lactobacillus casei BL23, there are two genes, bnaG and manA, encoding a beta-N-acetylglucosaminidase precursor and a mannose-6-phosphate isomerase, respectively. In this work, we show that L. casei is able to grow in the presence of lacto-N-triose as a carbon source. Inactivation of bnaG abolished the growth of L. casei on this oligosaccharide, demonstrating that BnaG is involved in its metabolism. Interestingly, whole cells of a bnaG mutant were totally devoid of beta-N-acetylglucosaminidase activity, suggesting that BnaG is an extracellular wall-attached enzyme. In addition to hydrolyzing lacto-N-triose into N-acetylglucosamine and lactose, the purified BnaG enzyme also catalyzed the hydrolysis of 3'-N-acetylglucosaminyl-mannose and 3'-N-acetylgalactosaminyl-galactose. L. casei can be cultured in the presence of 3'-N-acetylglucosaminyl-mannose as a carbon source, but, curiously, the bnaG mutant strain was not impaired in its utilization. These results indicate that the assimilation of 3'-N-acetylglucosaminyl-mannose is independent of BnaG. Enzyme activity and growth analysis with a manA-knockout mutant showed that ManA is involved in the utilization of the mannose moiety of 3'-N-acetylglucosaminyl-mannose. Here we describe the physiological role of a beta-N-acetylglucosaminidase in lactobacilli, and it supports the metabolic adaptation of L. casei to the N-acetylglucosaminide-rich gut niche.