ArticlePloS one2012
Fine tuning of the lactate and diacetyl production through promoter engineering in Lactococcus lactis.
Article in PloS one, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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Who cites it
29 citing papers in PubMed, 78 citations in OpenAlex.
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- New Effective Method of Lactococcus Genome Editing Using Guide RNA-Directed Transposition.International journal of molecular sciences · 2022Article
- Development of a novel expression system in lactic acid bacteria controlled by a broad-host-range promoter PMicrobial cell factories · 2022Article
- Fermentation of Agri-Food Waste: A Promising Route for the Production of Aroma Compounds.Foods (Basel, Switzerland) · 2021Review
- Identification and characterization of a moonlighting protein-enolase for surface display in Streptococcus thermophilus.Microbial cell factories · 2020Article
- Efficient production of α-acetolactate by whole cell catalytic transformation of fermentation-derived pyruvate.Microbial cell factories · 2019Article
- Beverage and Food Fragrance Biotechnology, Novel Applications, Sensory and Sensor Techniques: An Overview.Foods (Basel, Switzerland) · 2019Review
- Targeted and Repetitive Chromosomal Integration Enables High-Level Heterologous Gene Expression inApplied and environmental microbiology · 2019Article
- Systems Biology - A Guide for Understanding and Developing Improved Strains of Lactic Acid Bacteria.Frontiers in microbiology · 2019Review
- Redox cofactor engineering in industrial microorganisms: strategies, recent applications and future directions.Journal of industrial microbiology & biotechnology · 2018Review
- A Cytoplasmic Antiholin Is Embedded In Frame with the Holin in a Lactobacillus fermentum Bacteriophage.Applied and environmental microbiology · 2018Article
- Characterization of aBiotechnology reports (Amsterdam, Netherlands) · 2018Article
- A Single-Plasmid Genome Editing System for Metabolic Engineering ofFrontiers in microbiology · 2018Article
- Development of a counterselectable seamless mutagenesis system in lactic acid bacteria.Microbial cell factories · 2017Article
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Authors and funding
5 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Lactococcus lactis is a well-studied bacterium widely used in dairy fermentation and capable of producing metabolites with organoleptic and nutritional characteristics. For fine tuning of the distribution of glycolytic flux at the pyruvate branch from lactate to diacetyl and balancing the production of the two metabolites under aerobic conditions, a constitutive promoter library was constructed by randomizing the promoter sequence of the H(2)O-forming NADH oxidase gene in L. lactis. The library consisted of 30 promoters covering a wide range of activities from 7,000 to 380,000 relative fluorescence units using a green fluorescent protein as reporter. Eleven typical promoters of the library were selected for the constitutive expression of the H(2)O-forming NADH oxidase gene in L. lactis, and the NADH oxidase activity increased from 9.43 to 58.17-fold of the wild-type strain in small steps of activity change under aerobic conditions. Meanwhile, the lactate yield decreased from 21.15 ± 0.08 mM to 9.94 ± 0.07 mM, and the corresponding diacetyl production increased from 1.07 ± 0.03 mM to 4.16 ± 0.06 mM with the intracellular NADH/NAD(+) ratios varying from 0.711 ± 0.005 to 0.383 ± 0.003. The results indicated that the reduced pyruvate to lactate flux was rerouted to the diacetyl with an almost linear flux variation via altered NADH/NAD(+) ratios. Therefore, we provided a novel strategy to precisely control the pyruvate distribution for fine tuning of the lactate and diacetyl production through promoter engineering in L. lactis. Interestingly, the increased H(2)O-forming NADH oxidase activity led to 76.95% lower H(2)O(2) concentration in the recombinant strain than that of the wild-type strain after 24 h of aerated cultivation. The viable cells were significantly elevated by four orders of magnitude within 28 days of storage at 4°C, suggesting that the increased enzyme activity could eliminate H(2)O(2) accumulation and prolong cell survival.
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