ArticleBiotechnology for biofuels and bioproducts2023
Sustainable and high-level microbial production of plant hemoglobin in Corynebacterium glutamicum.
Article in Biotechnology for biofuels and bioproducts, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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8 citing papers in PubMed, 21 citations in OpenAlex.
- Putative membrane protein Cgl2168 and its variant enhance thermotolerance inSynthetic and systems biotechnology · 2026Article
- Cell-Free Production of Soybean Leghemoglobins and Nonsymbiotic Hemoglobin.ACS synthetic biology · 2025Article
- Secretory Production of Plant Heme-Containing Globins by Recombinant Yeast via Precision Fermentation.Foods (Basel, Switzerland) · 2025Article
- Automated characterization and analysis of expression compatibility between regulatory sequences and metabolic genes inSynthetic and systems biotechnology · 2024Article
- Multi-modular metabolic engineering of heme synthesis inSynthetic and systems biotechnology · 2024Article
- How Nitric Oxide Hindered the Search for Hemoglobin-Based Oxygen Carriers as Human Blood Substitutes.International journal of molecular sciences · 2023Review
- Biosynthesis of High-Active Hemoproteins by the Efficient Heme-Supply Pichia Pastoris Chassis.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2023Article
- High-level expression of leghemoglobin inFrontiers in bioengineering and biotechnology · 2023Article
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Authors and funding
14 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundPlant hemoglobin shows great potential as a food additive to circumvent the controversy of using animal materials. Microbial fermentation with engineered microorganisms is considered as a promising strategy for sustainable production of hemoglobin. As an endotoxin-free and GRAS (generally regarded as safe) bacterium, Corynebacterium glutamicum is an attractive host for hemoglobin biosynthesis.
resultsHerein, C. glutamicum was engineered to efficiently produce plant hemoglobin. Hemoglobin genes from different sources including soybean and maize were selected and subjected to codon optimization. Interestingly, some candidates optimized for the codon usage bias of Escherichia coli outperformed those for C. glutamicum regarding the heterologous expression in C. glutamicum. Then, saturated synonymous mutation of the N-terminal coding sequences of hemoglobin genes and fluorescence-based high-throughput screening produced variants with 1.66- to 3.45-fold increase in hemoglobin expression level. To avoid the use of toxic inducers, such as isopropyl-β-D-thiogalactopyranoside, two native inducible expression systems based on food additives propionate and gluconate were developed. Promoter engineering improved the hemoglobin expression level by 2.2- to 12.2-fold. Combination of these strategies and plasmid copy number modification allowed intracellular production of hemoglobin up to approximately 20% of total protein. Transcriptome and proteome analyses of the hemoglobin-producing strain revealed the cellular response to excess hemoglobin accumulation. Several genes were identified as potential targets for further enhancing hemoglobin production.
conclusionsIn this study, production of plant hemoglobin in C. glutamicum was systematically engineered by combining codon optimization, promoter engineering, plasmid copy number modification, and multi-omics-guided novel target discovery. This study offers useful design principles to genetically engineer C. glutamicum for the production of hemoglobin and other recombinant proteins.
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