A regulatory gene (<i>ECO‐orf4</i>) required for ECO‐0501 biosynthesis in <i>Amycolatopsis orientalis</i>

Shen, Yang; Huang, He; Zhu, Linghua; Luo, Mao; Chen, Daijie

doi:10.1002/jobm.201200314

Cited by 5 publications

(11 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To establish a conjunction system, an E. coli‐Amycolatopsis shuttle vector pLYZW7‐3 was constructed by cloning 2830 bp of the region necessary for replication into pLYZL102. Plasmid pLYZL102 was constructed in our previous work and included the ori T fragment required for conjugal transfer from E. coli to Amycolatopsis . Another gene, aac(3)IV from pLYZL102, encoded resistance to apramycin and served as the selection marker.…”

Section: Resultsmentioning

confidence: 99%

Characterization of plasmid pXL100 from Amycolatopsis orientalis HCCB10007 and construction of a shuttle vector

Zhu

et al. 2014

J. Basic Microbiol.

Self Cite

View full text Add to dashboard Cite

Many strains of Amycolatopsis, such as Amycolatopsis orientalis, A. balhimycina, and A. mediterranei, are important antibiotic producers. Three indigenous plasmids, pMEA100, pMEA300, and pA387, found in this genus have been sequenced. However, only some vectors based on pA387 have been widely applied in Amycolatopsis research. An indigenous plasmid, pXL100, was isolated from the vancomycin producer A. orientalis HCCB10007. Sequence analysis of pXL100 revealed its total length to be 33,499 bp and GC content to be 68.9%. A 2830-bp fragment containing three ORFs has been identified as essential for replication in A. orientalis, but it has no significant similarity to any known replicons. A vector, pLYZW7-3, was constructed based on the pXL100 replicon and could be transferred into A. mediterranei and A. orientalis by electroporation or conjugation with high frequency. A mutant with a disrupted gene was successfully complemented with the pLYZW7-3 vector, indicating that the vector is potentially useful in Amycolatopsis research.

show abstract

Section: Resultsmentioning

confidence: 99%

Characterization of plasmid pXL100 from Amycolatopsis orientalis HCCB10007 and construction of a shuttle vector

Zhu

et al. 2014

J. Basic Microbiol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…All A. orientalis strains were grown on Gauze's synthetic agar medium (GM) for sporulation, cultivated at 28 °C with agitation at 220 rpm in shake‐flasks that contained seed medium for vegetative growth. Seed cultures were then transferred into shake‐flasks containing F1 , F2 , or F5 medium (60 g L −1 glycerol, 20 g L −1 yeast extract, 6 g L −1 KNO 3 , 0.2 g L −1 KH 2 PO 4 , 0.4 g L −1 MgCl 2 . 6H 2 O, 3 g L −1 CaCO 3 , pH 6.5), which were then incubated for 5 days to produce different products, that is, vancomycin, ECO‐0501, and coproporphyrin.…”

Section: Methodsmentioning

confidence: 99%

“…To exclude the possibility that hemA2 deletion led to a polar effect on the expression of other ECO‐0501 biosynthetic genes, the transcriptions of the downstream genes ORF 17 and ORF 18 of hemA2 were analyzed by reverse transcription PCR. RNA isolation and transcriptional analyses were performed as previously described .…”

Section: Methodsmentioning

confidence: 99%

“…The fermentation broth was extracted with methanol to yield a crude organic extract containing ECO‐0501, coproporphyrin, or vancomycin. HPLC methods for the separation of ECO‐0501 and vancomycin were conducted as previously described . Coproporphyrin production was assessed by HPLC using an Agilent Poroshell 120 EC‐C18 column (2.7 µm, 4.6 × 150 mm) at 390 nm with a flow rate 0.7 ml min −1 .…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Role of two 5‐aminolevulinic acid biosynthetic pathways in heme and secondary metabolite biosynthesis in Amycolatopsis orientalis

et al. 2017

Self Cite

View full text Add to dashboard Cite

Analysis of the Amycolatopsis orientalis genome revealed that two genes, hemA1 and hemA2, belonging to divergent pathways, were involved in the biosynthesis of 5-aminolevulinic acid. The roles of hemA1 and hemA2 were elucidated via genetic manipulation and metabolite analysis. The disruption of hemA1, encoding the glutamyl-tRNA reductase of the C pathway, was essential for cell growth and is used for heme synthesis. Overexpression of hemA1 resulted in elevated vancomycin and ECO-0501 production in Amycolatopsis orientalis, and it was also effective in increasing the production of daptomycin and natamycin in other Streptomycetes. The disruption of hemA2 indicated that it encodes the 5-aminolevulinic acid synthase of the Shemin pathway, serving as a key enzyme for the synthesis of the precursor aminohydroxycyclopentenone unit of ECO-0501. However, hemA2 disruption could not be complemented by the addition of 5-aminolevulinic acid or by the expression of hemA2 outside of the ECO-0501 gene cluster. The synthesis of ECO-0501 was only restored by the insertion of hemA2 at its original locus. The hemA2 gene could partly complement the hemA1 deficiency. Overexpression of hemA1, a key gene from the heme biosynthetic pathway, is proposed here as a new approach to improve the production of secondary metabolites in bacteria, whereas hemA2 plays different roles depending on its pattern of expression.

show abstract

“…The sugar oxidoreductase, encoded by ORF 13, oxidizes d ‐glucose to form a d ‐glucuronic acid, which is subsequently transferred onto a hydroxyl group of the polyketide chain by a glycosyltransferase (ORF 14) . A positive regulatory gene ( ECO‐orf4 ) was found by genetic studies to be essential for ECO‐0501 biosynthesis …”

Section: Polyketides Biosynthesis In Amycolatopsismentioning

confidence: 99%

Progress in Understanding the Genetic Information and Biosynthetic Pathways behind Amycolatopsis Antibiotics, with Implications for the Continued Discovery of Novel Drugs

Shen

et al. 2015

ChemBioChem

View full text Add to dashboard Cite

Species of Amycolatopsis, well recognized as producers of both vancomycin and rifamycin, are also known for producing other secondary metabolites, with wide usage in medicine and agriculture. The molecular genetics of natural antibiotics produced by this genus have been well studied. Since the rise of antibiotic resistance, finding new drugs to fight infection has become an urgent priority. Progress in understanding the biosynthesis of metabolites greatly helps the rational manipulation of biosynthetic pathways, and thus to achieve the goal of generating novel natural antibiotics. The efforts made in exploiting Amycolatopsis genome sequences for the discovery of novel natural products and biosynthetic pathways are summarized.

show abstract

A regulatory gene (ECO‐orf4) required for ECO‐0501 biosynthesis in Amycolatopsis orientalis

Cited by 5 publications

References 13 publications

Characterization of plasmid pXL100 from Amycolatopsis orientalis HCCB10007 and construction of a shuttle vector

Characterization of plasmid pXL100 from Amycolatopsis orientalis HCCB10007 and construction of a shuttle vector

Role of two 5‐aminolevulinic acid biosynthetic pathways in heme and secondary metabolite biosynthesis in Amycolatopsis orientalis

Progress in Understanding the Genetic Information and Biosynthetic Pathways behind Amycolatopsis Antibiotics, with Implications for the Continued Discovery of Novel Drugs

Contact Info

Product

Resources

About