| 1. |
Yohei Ishibashi, Yusuke Nagamatsu, Sandra Meyer, Nozomu Okino, Rudolf Geyer, Makoto Ito, Transglycosylation-based fluorescent labeling of glycolipids by EGALC, CHEMISTRY AND PHYSICS OF LIPIDS, 10.1016/j.chemphyslip.2009.06.052, Vol.160, p.S35, 2009.08. |
| 2. |
Yohei Ishibashi, Functions and applications of glycolipid-hydrolyzing microbial glycosidases, Bioscience, Biotechnology and Biochemistry, 10.1093/bbb/zbac089, Vol.86, No.8, pp.974-984, 2022.08, Glycolipids are important components of cell membranes in several organisms. The major glycolipids in mammals are glycosphingolipids (GSLs), which are composed of ceramides. In mammals, GSLs are degraded stepwise from the non-reducing end of the oligosaccharides via exo-type glycosidases. However, endoglycoceramidase (EGCase), an endo-type glycosidase found in actinomycetes, is a unique enzyme that directly acts on the glycosidic linkage between oligosaccharides and ceramides to generate intact oligosaccharides and ceramides. Three molecular species of EGCase, namely EGCase I, EGCase II, and endogalactosylceramidase, have been identified based on their substrate specificity. EGCrP1 and EGCrP2, which are homologs of EGCase in pathogenic fungi, were identified as the first fungal glucosylceramide- and sterylglucoside-hydrolyzing glycosidases, respectively. These enzymes are promising targets for antifungal drugs against pathogenic fungi. This review describes the functions and properties of these microbial glycolipid-degrading enzymes, the molecular basis of their differential substrate specificity, and their applications.. |
| 3. |
Makoto Ito, Yohei Ishibashi, Takashi Watanabe, Jun Iwaki, Toyohisa Kurita, Nozomu Okino , Assays and Utilization of Enzymes Involved in Glycolipid Metabolism in Bacteria and Fungi, Methods in Molecular Biology, https://doi.org/10.1007/978-1-0716-2910-9_18, 2023.01. |
| 4. |
Takashi Watanabe, Yohei Ishibashi, Makoto Ito, Physiological significance of glycolipid catabolism in Cryptococcus neoformans, Trends in Glycoscience and Glycotechnology, 10.4052/tigg.1504.1E, 2015.09, The pathogenic fungus Cryptococcus neoformans causes cryptococcosis, an opportunistic infectious disease resulting in 600,000 deaths per year. The two major glycolipids in C. neoformans are glucosylceramide (GlcCer) with a fungus-specific ceramide (methyl d18 : 2/h18 : 0) and ergosteryl β-glucoside (EG); however, the catabolic pathway of these glycolipids has not yet been uncovered. We found two homologues of endoglycoceramidase (EGCase, EC 3.2.1.123) in C. neoformans, designated Endoglycoceramidase- related Protein 1 and 2 (EGCrP1 and EGCrP2). EGCase hydrolyzes the O-glycosidic linkage between oligosaccharides and ceramides in various glycosphingolipids. However, EGCrP1 and EGCrP2 show completely different specificities; that is, EGCrP1 is a neutral glucocerebrosidase specific to GlcCer, whereas EGCrP2 is an acid β-glucosidase capable of hydrolyzing not only GlcCer but also various β-glucosides, including pNP β-glucoside and EG. Using each disruption mutant of egcrp1 and egcrp2, we elucidated that EGCrP1 plays an integral role in quality control of the fungus-specific GlcCer by eliminating immature GlcCer, which are byproducts of the GlcCer synthesis pathway, whereas EGCrP2 is involved in the catabolism of EG in the vacuoles of C. neoformans. The analysis of egcrp1-disrupted mutants also revealed that the quality control of fungus-specific GlcCer is strongly linked to the formation of the polysaccharide capsule, an important virulence factor. On the other hand, the disruption of EG catabolism resulted in growth arrest, dysfunction in cell budding, and abnormal vacuole morphology. These results indicate that catabolism of two different glycolipids plays different physiological roles in C. neoformans and strongly suggest EGCrP1 and EGCrP2 as targets for anticryptococcal drugs with a new mechanism of action.. |
| 5. |
Yohei Ishibashi, Virus-induced metabolic rewiring of a marine algal glycosphingolipid biosynthesis pathway during infection, Trends in Glycoscience and Glycotechnology, 10.4052/tigg.1607.6E, 2016.08. |
| 6. |
Yohei Ishibashi, Naked mole-rats survive anoxic environment using fructose-driven glycolysis, Trends in Glycoscience and Glycotechnology, 10.4052/tigg.1715.6E, 2017.09. |
