Molecular cloning of the gene for the key carbocycle-forming enzyme in the biosynthesis of 2-deoxystreptamine-containing aminocyclitol antibiotics and its comparison with dehydroquinate synthase.
Molecular cloning of the gene for the key carbocycle-forming enzyme in the biosynthesis of 2-deoxystreptamine-containing aminocyclitol antibiotics and its comparison with dehydroquinate synthase.
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含2-脱氧链霉胺氨基环醇抗生素生物合成中关键碳环形成酶基因的分子克隆及其与脱氢奎宁酸合成酶的比较。
DOI：
10.7164/antibiotics.52.559
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
K. Kakinuma
中科院分区：
文献类型：
--
作者：
F. Kudo;H. Tamegai;T. Fujiwara;Uno Tagami;K. Hirayama;K. Kakinuma
2-脱氧链霉胺糖苷配基是在氨基环醇抗生素（包括新霉素、卡那霉素、妥布霉素、庆大霉素、西索米星、布替罗新和核糖霉素）中发现的常见结构特征。参与2-脱氧链霉胺部分的生物合成的关键酶是2-脱氧-鲨肌醇（DOI）合酶，其催化从D-葡萄糖-6-磷酸到2-脱氧-鲨肌醇的碳环形成。最近从环状芽孢杆菌中成功分离到2-脱氧-海蟹肌醇合酶，促使我们利用反向遗传学方法克隆负责这一重要酶的基因。借助基于纯化的该酶42 kDa亚基的氨基末端序列构建的DNA探针，成功克隆了负责基因btrC。随后，btrC基因在大肠杆菌中异源表达，并通过化学分析证实重组多肽的2-脱氧-蟹肌醇合酶活性。btrC基因编码由368个氨基酸组成的蛋白质，分子量为40.7 kDa。我们先前提出的2-脱氧-青蟹-肌醇合酶与脱氢奎尼酸合酶的相似性已在它们的氨基酸序列的基础上得到证实。然而，也可以观察到序列的显著差异，特别是在关键的底物识别区域。BtrC序列与其他相关微生物产物的生物合成酶的序列的比较也进行了讨论。
The 2-deoxystreptamine aglycon is a common structural feature found in aminocyclitol antibiotics including neomycin, kanamycin, tobramycin, gentamicin, sisomicin, butirosin and ribostamycin. A key enzyme involved in the biosynthesis of the 2-deoxystreptamine moiety is 2-deoxy-scyllo-inosose (DOI) synthase which catalyses the carbocycle formation from D-glucose-6-phosphate to 2-deoxy-scyllo-inosose. The recent success of isolating the 2-deoxy-scyllo-inosose synthase from Bacillus circulans prompted us to clone the gene responsible for this important enzyme by the use of reverse genetics approach. With the aid of DNA probes constructed on the basis of the amino-terminal sequence of the purified 42 kDa subunit of the enzyme, the responsible gene btrC was successfully cloned. Subsequently the btrC gene was heterologously expressed in Escherichia coli, and the 2-deoxy-scyllo-inosose synthase activity of the recombinant polypeptide was confirmed by chemical analysis. The btrC gene encodes a protein composed of 368 amino acids with a molecular mass of 40.7 kDa. Our previous proposal for the similarity of 2-deoxy-scyllo-inosose synthase to dehydroquinate synthase has been confirmed on the basis of their amino acid sequences. Significant differences in the sequences can also be observed however, particularly in the crucial substrate recognition regions. Comparison of the BtrC sequence with those of biosynthetic enzymes for other related microbial products is also discussed.
影响因子：
--
作者：
August, PR;Tang, L;Floss, HG
通讯作者：
Floss, HG
DOI：
10.1002/bms.1200160119
发表时间：
1988-10-01
期刊：
BIOMEDICAL AND ENVIRONMENTAL MASS SPECTROMETRY
影响因子：
--
作者：
BIEMANN, K
通讯作者：
BIEMANN, K