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首页> 外文期刊>The Journal of Antibiotics: An International Journal >Cloning of the Pactamycin Biosynthetic Gene Cluster and Characterization of a Crucial Glycosyltransferase Prior to a Unique Cyclopentane Ring Formation
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Cloning of the Pactamycin Biosynthetic Gene Cluster and Characterization of a Crucial Glycosyltransferase Prior to a Unique Cyclopentane Ring Formation

机译：Pactamycin生物合成基因簇的克隆和关键糖基转移酶的表征，然后形成独特的环戊烷环

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摘要

The biosynthetic gene (pct) cluster for an antitumor antibiotic pactamycin was identified by use of a gene for putative radical S-adenosylmethionine methyltransferase as a probe.The pet gene cluster is localized to a 34 kb contiguous DNA from Streptomyces pactum NBRC 13433 and contains 24 open reading frames.Based on the bioinformatic analysis,a plausible biosynthetic pathway for pactamycin comprising of a unique cyclopentane ring,3-aminoacetophenone,and 6-methylsalicylate was proposed.The pctL gene encoding a glycosyltransferase was speculated to be involved in an N-glycoside formation between 3-aminoacetophenone and UDP-N-acetyl-alpha-D-glucosamine prior to a unique cyclopentane ring formation.The pctL gene was then heterologously expressed in Escherichia coli and the enzymatic activity of the recombinant PctL protein was investigated.Consequently,the PctL protein was found to catalyze the expected reaction forming beta-N-glycoside.The enzymatic activity of the PctL protein clearly confirmed that the present identified gene cluster is for the biosynthesis of pactamycin.Also,a glycosylation prior to cyclopentane ring formation was proposed to be a general strategy in the biosynthesis of the structurally related cyclopentane containing compounds.
机译：通过使用推定的自由基S-腺苷甲硫氨酸甲基转移酶基因作为探针鉴定抗肿瘤抗生素pactamycin的生物合成基因（pct）宠物基因簇定位于来自链霉菌NBtum 13433的34 kb连续DNA中，包含24在生物信息学分析的基础上，提出了一种由独特的环戊烷环，3-氨基苯乙酮和6-甲基水杨酸酯组成的pactamycin的生物合成途径。推测编码糖基转移酶的pctL基因与N-糖苷有关。在独特的环戊烷环形成之前先在3-氨基苯乙酮和UDP-N-乙酰基-α-D-葡萄糖胺之间形成PtL基因，然后在大肠杆菌中异源表达pctL基因，并研究重组PctL蛋白的酶促活性。发现PctL蛋白可以催化预期的反应形成β-N-糖苷.PctL蛋白的酶促活性明显因此，有人提出在环戊烷环形成之前进行糖基化是生物合成结构相关的含环戊烷的化合物的一般策略。

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