Genetic engineering and heterologous expression of the disorazol biosynthetic gene cluster via Red/ET recombineering

Qiang Tu; Jennifer Herrmann; Shengbiao Hu; Ritesh Raju; Xiaoying Bian; Youming Zhang; Rolf Muller

Genetic engineering and heterologous expression of the disorazol biosynthetic gene cluster via Red/ET recombineering

Qiang Tu, Jennifer Herrmann, Shengbiao Hu, Ritesh Raju, Xiaoying Bian, Youming Zhang, Rolf Muller

Research output: Contribution to journal › Article › peer-review

32 Citations (Scopus)

Abstract

Disorazol, a macrocyclic polykitide produced by the myxobacteriumSorangium cellulosum So ce12 and it is reported to have potential cytotoxic activity towards several cancer cell lines, including multi-drug resistant cells. The disorazol biosynthetic gene cluster (dis) fromSorangium cellulosum (So ce12) was identified by transposon mutagenesis and cloned in a bacterial artificial chromosome (BAC) library. The 58-kb dis core gene cluster was reconstituted from BACs via Red/ET recombineering and expressed in Myxococcus xanthusDK1622. For the first time ever, a myxobacterial trans-AT polyketide synthase has been expressed heterologously in this study. Expression inM. xanthus allowed us to optimize the yield of several biosynthetic products using promoter engineering. The insertion of an artificial synthetic promoter upstream of the disD gene encoding a discrete acyl transferase (AT), together with an oxidoreductase (Or), resulted in 7-fold increase in disorazol production. The successful reconstitution and expression of the genetic sequences encoding for these promising cytotoxic compounds will allow combinatorial biosynthesis to generate novel disorazol derivatives for further bioactivity evaluation.

Original language	English
Article number	21066
Number of pages	10
Journal	Scientific Reports
Volume	6
Publication status	Published - 15 Feb 2016

Bibliographical note

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Keywords

antibiotics
genes
peptides

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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title = "Genetic engineering and heterologous expression of the disorazol biosynthetic gene cluster via Red/ET recombineering",

abstract = "Disorazol, a macrocyclic polykitide produced by the myxobacteriumSorangium cellulosum So ce12 and it is reported to have potential cytotoxic activity towards several cancer cell lines, including multi-drug resistant cells. The disorazol biosynthetic gene cluster (dis) fromSorangium cellulosum (So ce12) was identified by transposon mutagenesis and cloned in a bacterial artificial chromosome (BAC) library. The 58-kb dis core gene cluster was reconstituted from BACs via Red/ET recombineering and expressed in Myxococcus xanthusDK1622. For the first time ever, a myxobacterial trans-AT polyketide synthase has been expressed heterologously in this study. Expression inM. xanthus allowed us to optimize the yield of several biosynthetic products using promoter engineering. The insertion of an artificial synthetic promoter upstream of the disD gene encoding a discrete acyl transferase (AT), together with an oxidoreductase (Or), resulted in 7-fold increase in disorazol production. The successful reconstitution and expression of the genetic sequences encoding for these promising cytotoxic compounds will allow combinatorial biosynthesis to generate novel disorazol derivatives for further bioactivity evaluation.",

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T1 - Genetic engineering and heterologous expression of the disorazol biosynthetic gene cluster via Red/ET recombineering

AU - Tu, Qiang

AU - Herrmann, Jennifer

AU - Hu, Shengbiao

AU - Raju, Ritesh

AU - Bian, Xiaoying

AU - Zhang, Youming

AU - Muller, Rolf

PY - 2016/2/15

Y1 - 2016/2/15

N2 - Disorazol, a macrocyclic polykitide produced by the myxobacteriumSorangium cellulosum So ce12 and it is reported to have potential cytotoxic activity towards several cancer cell lines, including multi-drug resistant cells. The disorazol biosynthetic gene cluster (dis) fromSorangium cellulosum (So ce12) was identified by transposon mutagenesis and cloned in a bacterial artificial chromosome (BAC) library. The 58-kb dis core gene cluster was reconstituted from BACs via Red/ET recombineering and expressed in Myxococcus xanthusDK1622. For the first time ever, a myxobacterial trans-AT polyketide synthase has been expressed heterologously in this study. Expression inM. xanthus allowed us to optimize the yield of several biosynthetic products using promoter engineering. The insertion of an artificial synthetic promoter upstream of the disD gene encoding a discrete acyl transferase (AT), together with an oxidoreductase (Or), resulted in 7-fold increase in disorazol production. The successful reconstitution and expression of the genetic sequences encoding for these promising cytotoxic compounds will allow combinatorial biosynthesis to generate novel disorazol derivatives for further bioactivity evaluation.

AB - Disorazol, a macrocyclic polykitide produced by the myxobacteriumSorangium cellulosum So ce12 and it is reported to have potential cytotoxic activity towards several cancer cell lines, including multi-drug resistant cells. The disorazol biosynthetic gene cluster (dis) fromSorangium cellulosum (So ce12) was identified by transposon mutagenesis and cloned in a bacterial artificial chromosome (BAC) library. The 58-kb dis core gene cluster was reconstituted from BACs via Red/ET recombineering and expressed in Myxococcus xanthusDK1622. For the first time ever, a myxobacterial trans-AT polyketide synthase has been expressed heterologously in this study. Expression inM. xanthus allowed us to optimize the yield of several biosynthetic products using promoter engineering. The insertion of an artificial synthetic promoter upstream of the disD gene encoding a discrete acyl transferase (AT), together with an oxidoreductase (Or), resulted in 7-fold increase in disorazol production. The successful reconstitution and expression of the genetic sequences encoding for these promising cytotoxic compounds will allow combinatorial biosynthesis to generate novel disorazol derivatives for further bioactivity evaluation.

KW - antibiotics

KW - genes

KW - peptides

UR - http://handle.uws.edu.au:8081/1959.7/uws:33884

M3 - Article

SN - 2045-2322

VL - 6

JO - Scientific Reports

JF - Scientific Reports

M1 - 21066

ER -

Genetic engineering and heterologous expression of the disorazol biosynthetic gene cluster via Red/ET recombineering

Abstract

Bibliographical note

Open Access - Access Right Statement

Keywords

UN SDGs

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