Use of Transposon Promoter-Probe Vectors in the Metabolic Engineering of the Obligate Methanotroph Methylomonas sp. Strain 16a for Enhanced C 40 Carotenoid Synthesis
| Autor: | Melissa D. Bosak, Qiong Cheng, Kyle Knoke, Pamela L. Sharpe, Deana J. Dicosimo, Luan Tao, Rick W. Ye |
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| Rok vydání: | 2007 |
| Předmět: |
DNA
Bacterial Transposable element Canthaxanthin Genetics and Molecular Biology Methylomonas Xanthophylls Biology Applied Microbiology and Biotechnology Genome Metabolic engineering chemistry.chemical_compound Astaxanthin Gene cluster Promoter Regions Genetic Molecular Biology Gene Recombination Genetic Genetics Ecology Chromosome Sequence Analysis DNA Chromosomes Bacterial biology.organism_classification Carotenoids chemistry Multigene Family DNA Transposable Elements Genome Bacterial Metabolic Networks and Pathways Food Science Biotechnology |
| Zdroj: | Applied and Environmental Microbiology. 73:1721-1728 |
| ISSN: | 1098-5336 0099-2240 |
| DOI: | 10.1128/aem.01332-06 |
| Popis: | The recent expansion of genetic and genomic tools for metabolic engineering has accelerated the development of microorganisms for the industrial production of desired compounds. We have used transposable elements to identify chromosomal locations in the obligate methanotroph Methylomonas sp. strain 16a that support high-level expression of genes involved in the synthesis of the C 40 carotenoids canthaxanthin and astaxanthin. with three promoterless carotenoid transposons, five chromosomal locations—the fliCS , hsdM , ccp-3 , cysH , and nirS regions—were identified. Total carotenoid synthesis increased 10- to 20-fold when the carotenoid gene clusters were inserted at these chromosomal locations compared to when the same carotenoid gene clusters were integrated at neutral locations under the control of the promoter for the gene conferring resistance to chloramphenicol. A chromosomal integration system based on sucrose lethality was used to make targeted gene deletions or site-specific integration of the carotenoid gene cluster into the Methylomonas genome without leaving genetic scars in the chromosome from the antibiotic resistance genes that are present on the integration vector. The genetic approaches described in this work demonstrate how metabolic engineering of microorganisms, including the less-studied environmental isolates, can be greatly enhanced by identifying integration sites within the chromosome of the host that permit optimal expression of the target genes. |
| Databáze: | OpenAIRE |
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