Journal of Applied Glycoscience
Online ISSN : 1880-7291
Print ISSN : 1344-7882
ISSN-L : 1344-7882
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  • Wataru Saburi, Tomoya Ota, Koji Kato, Takayoshi Tagami, Keitaro Yamash ...
    2023 Volume 70 Issue 2 Pages 43-52
    Published: May 20, 2023
    Released on J-STAGE: August 02, 2023
    Advance online publication: March 11, 2023

    β-Galactosidase (EC hydrolyzes β-D-galactosidic linkages at the non-reducing end of substrates to produce β-D-galactose. Lacticaseibacillus casei is one of the most widely utilized probiotic species of lactobacilli. It possesses a putative β-galactosidase belonging to glycoside hydrolase family 35 (GH35). This enzyme is encoded by the gene included in the gene cluster for utilization of lacto-N-biose I (LNB; Galβ1-3GlcNAc) and galacto-N-biose (GNB; Galβ1-3GalNAc) via the phosphoenolpyruvate: sugar phosphotransferase system. The GH35 protein (GnbG) from L. casei BL23 is predicted to be 6-phospho-β-galactosidase (EC However, its 6-phospho-β-galactosidase activity has not yet been examined, whereas its hydrolytic activity against LNB and GNB has been demonstrated. In this study, L. casei JCM1134 LBCZ_0230, homologous to GnbG, was characterized enzymatically and structurally. A recombinant LBCZ_0230, produced in Escherichia coli, exhibited high hydrolytic activity toward o-nitrophenyl β-D-galactopyranoside, p-nitrophenyl β-D-galactopyranoside, LNB, and GNB, but not toward o-nitrophenyl 6-phospho-β-D-galactopyranoside. Crystal structure analysis indicates that the structure of subsite −1 of LBCZ_0230 is very similar to that of Streptococcus pneumoniae β-galactosidase BgaC and not suitable for binding to 6-phospho-β-D-galactopyranoside. These biochemical and structural analyses indicate that LBCZ_0230 is a β-galactosidase. According to the prediction of LNB's binding mode, aromatic residues, Trp190, Trp240, Trp243, Phe244, and Tyr458, form hydrophobic interactions with N-acetyl-D-glucosamine residue of LNB at subsite +1.

  • Yuki Yoshitomi, Kiyoshi Kawai
    2023 Volume 70 Issue 2 Pages 53-58
    Published: May 20, 2023
    Released on J-STAGE: August 02, 2023
    Advance online publication: March 25, 2023
    Supplementary material

    The purpose of this study was to understand the effect of relative humidity (RH) on amylose-lipid complex (ALC) formation in amylose-lauric acid blend powder held at 50 °C (temperature slightly higher than the melting point of lauric acid) using differential scanning calorimetry (DSC) and X-ray diffraction. From DSC curves, the melting of crystalized lauric acid and two melting peaks of ALC were observed depending on RH. ALC formation was confirmed by X-ray diffraction pattern. The melting enthalpy (∆Hm) of lauric acid in the sample held at RH 0 % was lower than that of lauric acid only though there was no ALC formation. This suggests that crystallization of lauric acid was prevented by amylose. The ∆Hm of lauric acid increased with an increase in RH up to 79.0 % because liquid lauric acid would have fused as the result of enhanced repulsive force between liquid lauric acid and hydrated amylose. The ∆Hm of ALC increased with an increase in RH between 79.0 and 95.0 %. For ALC formation, amylose has to be mobile in the system, but dehydrated amylose is in a glassy (immobilize) state. According to the glass to rubber transition behavior of amorphous polymer, amylose held at 50 °C is suggested to become rubbery (mobile) state at RH 76.0 %. This interpretation will explain the reason why ALC formation began to be observed at the RH range between 72.4 and 79.0 %.