Reverse Engineering Targets for Recombinant Protein Production in Corynebacterium glutamicum Inspired by a Fast-Growing Evolved Descendant
| Autor: | Kyunghoon Park, Min Ju Lee, Jihoon Park, Jihyun F. Kim, Pil Kim |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
0301 basic medicine cellular energy Histology lcsh:Biotechnology Biomedical Engineering host for recombinant protein Bioengineering Biology medicine.disease_cause 01 natural sciences Corynebacterium glutamicum law.invention 03 medical and health sciences Plasmid law lcsh:TP248.13-248.65 010608 biotechnology medicine Derepression Original Research glucose consumption Genetics Mutation Strain (chemistry) Point mutation Structural gene Bioengineering and Biotechnology iron consumption 030104 developmental biology ribosome Recombinant DNA Biotechnology |
| Zdroj: | Frontiers in Bioengineering and Biotechnology Frontiers in Bioengineering and Biotechnology, Vol 8 (2020) |
| ISSN: | 2296-4185 |
| DOI: | 10.3389/fbioe.2020.588070 |
| Popis: | We previously reported a Corynebacterium glutamicum JH41 strain with a 58% faster growth rate through application of adaptive laboratory evolution. To verify that the fast-reproducing strain was useful as a host for recombinant protein expression, we introduced a plasmid responsible for the secretory production of a recombinant protein. The JH41 strain harboring the plasmid indeed produced the secretory recombinant protein at a 2.7-fold greater rate than its ancestral strain. To provide the reverse engineering targets responsible for boosting recombinant protein production and cell reproduction, we compared the genome sequence of the JH41 strain with its ancestral strain. Among the 15 genomic variations, a point mutation was confirmed in the 14 bases upstream of NCgl1959 (encoding a presumed siderophore-binding protein). This mutation allowed derepression of NCgl1959, thereby increasing iron consumption and ATP generation. A point mutation in the structural gene ramA (A239G), a LuxR-type global transcription regulator involved in central metabolism, allowed an increase in glucose consumption. Therefore, mutations to increase the iron and carbon consumption were concluded as being responsible for the enhanced production of recombinant protein and cell reproduction in the evolved host. |
| Databáze: | OpenAIRE |
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