β-[32P]UDP-N-乙酰葡糖胺的酶促合成
The enzymic synthesis of beta-[32P]UDP-N-acetylglucosamine.
作者信息
Heptinstall J, Ward P J, Hancock I C
机构信息
Microbiological Chemistry Research Laboratory, University of Newcastle upon Tyne, United Kingdom.
出版信息
Anal Biochem. 1978 Nov;91(1):158-65. doi: 10.1016/0003-2697(78)90826-6.
Cells of Micrococcus sp. 2102 incorporate inorganic [32P]phosphate from the medium into the sugar-phosphate polymer of the wall. Controlled acid hydrolysis of sodium dodecyl sulphate-extracted cells gives N-acetylglucosamine 6-[32P]-phosphate which can be purified by ion-exchange chromatography and incubated with UTP in the presence of crude preparations of phosphoacetylglucosamine mutase from Neurospora crassa and UTP:N-acetylglucosamine 1-phosphate phosphotransferase from Bacillus licheniformis which act in concert to synthesise beta-[32P]UDP-N-acetylglucosamine.
微球菌属2102菌株的细胞将培养基中的无机[32P]磷酸盐整合到细胞壁的糖磷酸盐聚合物中。对十二烷基硫酸钠提取的细胞进行受控酸水解，可得到N-乙酰葡糖胺6-[32P] - 磷酸盐，其可通过离子交换色谱法纯化，并在来自粗糙脉孢菌的磷酸乙酰葡糖胺变位酶粗制品和来自地衣芽孢杆菌的UTP：N-乙酰葡糖胺1-磷酸磷酸转移酶存在下与UTP一起孵育，这两种酶协同作用以合成β-[32P] UDP-N-乙酰葡糖胺。
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