Lacto-N-biosidase Encoded by a Novel Gene of Bifidobacterium longum Subspecies longum Shows Unique Substrate Specificity and Requires a Designated Chaperone for Its Active Expression

Sakurama, Haruko; Kiyohara, Masashi; Wada, Jun; Honda, Yoshiyuki; Yamaguchi, Masanori; Fukiya, Satoru; Yokota, Atsushi; Ashida, Hisashi; Kumagai, Hidehiko; Kitaoka, Motomitsu; Yamamoto, Kenji; Katayama, Takane

doi:10.1074/jbc.m113.484733

Cited by 83 publications

(76 citation statements)

References 45 publications

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“…In turn, the ␣-helices sit against the outer rim of domain II. Domain II is a (␤/␣) 8 barrel, or TIM barrel, which is not only the core fold of GH family 20 but is one of the most common folds among glycoside hydrolases in general (42). Domain III is all ␣-helical with four anti-parallel helices organized in a near planar arrangement.…”

Section: Volume 290 • Number 52 • December 25 2015mentioning

confidence: 99%

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A Second β-Hexosaminidase Encoded in the Streptococcus pneumoniae Genome Provides an Expanded Biochemical Ability to Degrade Host Glycans

Robb

Higgins

et al. 2015

Journal of Biological Chemistry

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Section: Volume 290 • Number 52 • December 25 2015mentioning

confidence: 99%

“…2B). The dimer interface, which exceeds 3000 Å 2 of buried surface area, is completed by loops extending from the (␤/␣) 8 barrel to interact with the same loops on the adjacent monomer, although this interaction contributes the minority of the dimer …”

Section: Volume 290 • Number 52 • December 25 2015mentioning

confidence: 99%

A Second β-Hexosaminidase Encoded in the Streptococcus pneumoniae Genome Provides an Expanded Biochemical Ability to Degrade Host Glycans

Robb

Higgins

et al. 2015

Journal of Biological Chemistry

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“…Glycoprotein degradation strategies have been examined among several bifidobacterial species (8,58), and EndoBI-1 homologs have been identified and characterized for strains of B. longum subsp. longum, B. breve, and others (8).…”

Section: Figmentioning

confidence: 99%

Oligosaccharides Released from Milk Glycoproteins Are Selective Growth Substrates for Infant-Associated Bifidobacteria

Karav

Parc

Bell

et al. 2016

Appl Environ Microbiol

128

117

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Milk, in addition to nourishing the neonate, provides a range of complex glycans whose construction ensures a specific enrichment of key members of the gut microbiota in the nursing infant, a consortium known as the milk-oriented microbiome. Milk glycoproteins are thought to function similarly, as specific growth substrates for bifidobacteria common to the breast-fed infant gut. Recently, a cell wall-associated endo-␤-N-acetylglucosaminidase (EndoBI-1) found in various infant-borne bifidobacteria was shown to remove a range of intact N-linked glycans. We hypothesized that these released oligosaccharide structures can serve as a sole source for the selective growth of bifidobacteria. We demonstrated that EndoBI-1 released N-glycans from concentrated bovine colostrum at the pilot scale. EndoBI-1-released N-glycans supported the rapid growth of Bifidobacterium longum subsp. infantis (B. infantis), a species that grows well on human milk oligosaccharides, but did not support growth of Bifidobacterium animalis subsp. lactis (B. lactis), a species which does not. Conversely, B. infantis ATCC 15697 did not grow on the deglycosylated milk protein fraction, clearly demonstrating that the glycan portion of milk glycoproteins provided the key substrate for growth. Mass spectrometry-based profiling revealed that B. infantis consumed 73% of neutral and 92% of sialylated N-glycans, while B. lactis degraded only 11% of neutral and virtually no (<1%) sialylated N-glycans. These results provide mechanistic support that N-linked glycoproteins from milk serve as selective substrates for the enrichment of infant-associated bifidobacteria capable of carrying out the initial deglycosylation. Moreover, released N-glycans were better growth substrates than the intact milk glycoproteins, suggesting that EndoBI-1 cleavage is a key initial step in consumption of glycoproteins. Finally, the variety of N-glycans released from bovine milk glycoproteins suggests that they may serve as novel prebiotic substrates with selective properties similar to those of human milk oligosaccharides. IMPORTANCEIt has been previously shown that glycoproteins serve as growth substrates for bifidobacteria. However, which part of a glycoprotein (glycans or polypeptides) is responsible for this function was not known. In this study, we used a novel enzyme to cleave conjugated N-glycans from milk glycoproteins and tested their consumption by various bifidobacteria. The results showed that the glycans selectively stimulated the growth of B. infantis, which is a key infant gut microbe. The selectivity of consumption of individual N-glycans was determined using advanced mass spectrometry (nano-liquid chromatography chip-quadrupole time of flight mass spectrometry [nano-LC-Chip-Q-TOF MS]) to reveal that B. infantis can consume the range of glycan structures released from whey protein concentrate. P rotein glycosylation is a common modification that adds a major dimension of complexity to protein structure and has been linked to significant roles in protein fun...

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“…Although the main function of LnbX must be the hydrolysis of lacto-N-tetraose (LNT: Galb1-3GlcNAcb1-3Galb1-4Glc) based on the catalytic efficiency for this substrate being relatively high (see Table 2), 19 the quite rare occurrence of LnbX homologues in nature and the distant relationship of LnbX with other proteins in the database prompted us to examine the heretofore unknown enzymatic activities of this enzyme. To this end, we examined the inhibitory effects of monosaccharides that constitute the sugar chains of host glycoproteins, glycolipids, and glycosaminoglycans, i.e., Fuc, Gal, GalNAc, Glc, GlcA, GlcNAc, Man, Neu5Ac, and Xyl.…”

Section: Inhibition Of Lnbx Activity In the Presence Of Galnacmentioning

confidence: 99%

“…Our previous studies demonstrated that GH20 LnbB exhibits strict specificity towards the unmodified LNB structure, while LnbX is capable of liberating GalNAcb1-3GlcNAc (a disaccharide structure found in O-mannosyl glycans of a-dystroglycan) 22 and 2 0 -fucosyl LNB (Fuca1-2Galb1-3GlcNAc) in addition to LNB. 18,19 LnbB has evolved from exo-b-N-acetylhexosaminidase, the main member of GH20, by acquiring the À2 subsite to accommodate the b-(1/3)-linked Gal residue of LNB. 23 GH20 lacto-Nbiosidase is distributed only in the Bifidobacterium and Streptomyces genera; nevertheless, these two bacterial groups likely evolved these enzymes differently because the amino acid sequence homology between these two enzymes is 38% at maximum.…”

Section: Introductionmentioning

confidence: 99%

Novel substrate specificities of two lacto-N-biosidases towards β-linked galacto-N-biose-containing oligosaccharides of globo H, Gb5, and GA1

Gotoh

Katoh

Sugiyama

et al. 2015

Carbohydrate Research

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Lacto-N-biosidase Encoded by a Novel Gene of Bifidobacterium longum Subspecies longum Shows Unique Substrate Specificity and Requires a Designated Chaperone for Its Active Expression

Cited by 83 publications

References 45 publications

A Second β-Hexosaminidase Encoded in the Streptococcus pneumoniae Genome Provides an Expanded Biochemical Ability to Degrade Host Glycans

A Second β-Hexosaminidase Encoded in the Streptococcus pneumoniae Genome Provides an Expanded Biochemical Ability to Degrade Host Glycans

Oligosaccharides Released from Milk Glycoproteins Are Selective Growth Substrates for Infant-Associated Bifidobacteria

Novel substrate specificities of two lacto-N-biosidases towards β-linked galacto-N-biose-containing oligosaccharides of globo H, Gb5, and GA1

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