Yokoyama K, Araki Y, Ito E
Eur J Biochem. 1987 May 15;165(1):47-53. doi: 10.1111/j.1432-1033.1987.tb11192.x.
The pathway for the de novo synthesis of a teichoic acid, poly(galactosylglycerol phosphate), in Bacillus coagulans AHU 1366 was studied by means of characterization and stepwise conversion of lipid-linked intermediates. Incubation of membranes with UDP-N-acetylglucosamine and UDP-glucose yielded a disaccharide-linked polyprenylpyrophosphate, whose sugar moiety was characterized as glucosyl(beta 1----4)N-acetylglucosamine (Glc-GlcNAc). By incubation with membranes and CDP-glycerol, Glc-GlcNAc-PP-prenol was converted to a series of glycolipids characterized as (Gro-P)1-6-Glc-GlcNAc-PP-prenol (Gro = glycerol). Glc-[14C]GlcNAc-PP-prenol was converted to polymer by incubation with membranes, CDP-glycerol and UDP-galactose. Smith degradation of the polymer gave two radioactive fragments corresponding to (Gro-P)3-Glc-GlcNAc and (Gro-P)4-Glc-GlcNAc. These results, together with data on gel chromatography of radioactive polymer synthesized from UDP-[3H]galactose, CDP-glycerol and Glc-[14C]GlcNAc-PP-prenol, led to the conclusion that in this strain poly(galactosylglycerol phosphate) is probably synthesized through the following pathway: GlcNAc-PP-prenol----Glc-GlcNAc-PP-prenol----(Gro-P)3-4 -Glc-GlcNAc-PP-prenol----(Gro-P-Gal)n- (Gro-P)3-4-Glc-GlcNAc-PP-prenol----(Gro-P-Gal)n- (Gro-P)3-4-Glc-GlcNAc-P-peptidoglycan complex.
通过对脂质连接中间体的表征和逐步转化,研究了凝结芽孢杆菌AHU 1366中磷壁酸——聚(半乳糖基甘油磷酸)的从头合成途径。用UDP-N-乙酰葡糖胺和UDP-葡萄糖孵育细胞膜,产生了一种二糖连接的聚异戊二烯焦磷酸,其糖部分被鉴定为葡萄糖基(β1→4)N-乙酰葡糖胺(Glc-GlcNAc)。通过与细胞膜和CDP-甘油一起孵育,Glc-GlcNAc-PP-异戊二烯醇被转化为一系列糖脂,其特征为(Gro-P)1-6-Glc-GlcNAc-PP-异戊二烯醇(Gro =甘油)。Glc-[14C]GlcNAc-PP-异戊二烯醇通过与细胞膜、CDP-甘油和UDP-半乳糖一起孵育转化为聚合物。该聚合物的史密斯降解产生了两个放射性片段,分别对应于(Gro-P)3-Glc-GlcNAc和(Gro-P)4-Glc-GlcNAc。这些结果,连同由UDP-[3H]半乳糖、CDP-甘油和Glc-[14C]GlcNAc-PP-异戊二烯醇合成的放射性聚合物的凝胶色谱数据,得出结论:在该菌株中,聚(半乳糖基甘油磷酸)可能通过以下途径合成:GlcNAc-PP-异戊二烯醇→Glc-GlcNAc-PP-异戊二烯醇→(Gro-P)3-4-Glc-GlcNAc-PP-异戊二烯醇→(Gro-P-Gal)n-(Gro-P)3-4-Glc-GlcNAc-PP-异戊二烯醇→(Gro-P-Gal)n-(Gro-P)3-4-Glc-GlcNAc-P-肽聚糖复合物。