Ashida H, Hayashi S, Sakamoto Y, Tsuji Y, Yamamoto K, Kumagai H, Tochikura T
Research Laboratory of Higashimaru Shoyu Co., Ltd., Hyogo, Japan.
Biosci Biotechnol Biochem. 1995 Nov;59(11):2028-32. doi: 10.1271/bbb.59.2028.
The actinomycete strain Streptomyces sp. H37 produces a novel glycosphingolipid-degrading enzyme. This strain was capable of converting ganglioside GM1 to lyso-GM1. After cultivation for 5 days in medium containing GM1, peptone, and detergent, GM1 was found to be almost completely converted to lyso-GM1. The product was purified on a DEAE-Sephadex A-25 column and thin layer chromatographies. The purified lyso-GM1 was hydrolyzed by endoglycoceramidase, and the released oligosaccharide moiety was identified as that of GM1 by HPLC using the pyridylaminoderivative method. The counterpart sphingosine moiety was confirmed with TLC. Moreover, the structure of lyso-GM1 was ascertained by 1H-NMR analysis. The maximum formation of lyso-GM1 was found in 50 mM potassium phosphate buffer (pH 7.5) containing 0.1% glycodeoxycholate. Various lyso-glycoshingolipids, including those of ganglio-, neolacto-, and globo-types, were formed from their parent glycosphingolipids using this strain.
放线菌菌株链霉菌属H37产生一种新型糖鞘脂降解酶。该菌株能够将神经节苷脂GM1转化为溶血GM1。在含有GM1、蛋白胨和去污剂的培养基中培养5天后,发现GM1几乎完全转化为溶血GM1。产物在DEAE-葡聚糖A-25柱和薄层色谱上进行纯化。纯化的溶血GM1被内切糖神经酰胺酶水解,释放的寡糖部分通过使用吡啶氨基衍生物法的HPLC鉴定为GM1的寡糖部分。鞘氨醇部分通过TLC得到确认。此外,溶血GM1的结构通过1H-NMR分析确定。在含有0.1%甘氨脱氧胆酸盐的50 mM磷酸钾缓冲液(pH 7.5)中发现溶血GM1的形成量最大。使用该菌株从其亲本糖鞘脂形成了各种溶血糖鞘脂,包括神经节型、新乳糖型和球型。