IDENTIFICATION OF DEBS-1, DEBS-2 AND DEBS-3, THE MULTIENZYME POLYPEPTIDES OF THE ERYTHROMYCIN-PRODUCING POLYKETIDE SYNTHASE FROM SACCHAROPOLYSPORA-ERYTHRAEA
IDENTIFICATION OF DEBS-1, DEBS-2 AND DEBS-3, THE MULTIENZYME POLYPEPTIDES OF THE ERYTHROMYCIN-PRODUCING POLYKETIDE SYNTHASE FROM SACCHAROPOLYSPORA-ERYTHRAEA
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DOI：
10.1016/0014-5793(92)80624-p
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发表时间：
1992-06-15
期刊：
影响因子：
3.5
通讯作者：
LEADLAY, PF
中科院分区：
文献类型：
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作者：
CAFFREY, P;BEVITT, DJ;LEADLAY, PF
此前已证明红色糖多孢菌的红霉素生物合成基因簇的ery A区域包含三个大的开放阅读框（ORF），编码6-脱氧红内酯B合酶（DEBS）的组分。 针对通过ORF 2和ORF 3基因的3'区的过表达获得的重组蛋白产生多克隆抗体。 在Western印迹实验中，每种抗血清与产红霉素的S.紫堇细胞 纯化这些推定的DEBS 2和DEBS 3蛋白并进行N-末端序列分析。 蛋白质序列与从ORF 2和3的DNA序列预测的翻译起始位点完全一致。 第三种高分子量蛋白质与DEBS 2和DEBS 3共纯化，其N-末端序列与从先前提出的ORF 1起始密码子上游约155个碱基对的DNA序列翻译的蛋白质序列相匹配。
The ery A region of the erythromycin biosynthetic gene cluster of Saccharopolyspora erythraea has previously been shown to contain three large open reading frames (ORFs) that encode the components of 6-deoxyerythronolide B synthase (DEBS). Polyclonal antibodies were raised against recombinant proteins obtained by overexpression of 3' regions of the ORF2 and ORF3 genes. In Western blotting experiments, each antiserum reacted strongly with a different high molecular weight protein in extracts of erythromycin-producing S. erythraea cells. These putative DEBS 2 and DEBS 3 proteins were purified and subjected to N-terminal sequence analysis. The protein sequences were entirely consistent with the translation start sites predicted from the DNA sequences of ORFs 2 and 3. A third high molecular weight protein co-purified with DEBS 2 and DEBS 3 and had an N-terminal sequence that matched a protein sequence translated from the DNA sequence some 155 base pairs upstream from the previously proposed start codon of ORF1.