Engineering Cyanobacteria To Synthesize and Export Hydrophilic Products

• Published in: Applied and Environmental Microbiology Vol. 76; no. 11; pp. 3462 - 3466
• Main Authors: Niederholtmeyer, Henrike, Wolfstädter, Bernd T, Savage, David F, Silver, Pamela A, Way, Jeffrey C
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.06.2010; American Society for Microbiology (ASM)
• Subjects: Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacteriology; Biological and medical sciences; Biological Transport; Biotechnology; Carbon; Carbon dioxide; Cyanobacteria; DNA, Bacterial - chemistry; DNA, Bacterial - genetics; Escherichia coli - growth & development; Escherichia coli - metabolism; Fructose - metabolism; Fundamental and applied biological sciences. Psychology; Gene expression; Genetic Engineering; Glucose; Glucose - metabolism; Gram-negative bacteria; Membrane Transport Proteins - genetics; Membrane Transport Proteins - metabolism; Microbiology; Molecular Sequence Data; Organic Chemicals - chemistry; Organic Chemicals - metabolism; Sequence Analysis, DNA; Synechococcus - genetics; Synechococcus - metabolism; Bacteria; Cyanobacteria
• ISSN: 0099-2240; 1098-5336; 1098-6596
• DOI: 10.1128/aem.00202-10

Metabolic engineering of cyanobacteria has the advantage that sunlight and CO₂ are the sole source of energy and carbon for these organisms. However, as photoautotrophs, cyanobacteria generally lack transporters to move hydrophilic primary metabolites across membranes. To address whether cyanobacteria could be engineered to produce and secrete organic primary metabolites, Synechococcus elongatus PCC7942 was engineered to express genes encoding an invertase and a glucose facilitator, which mediated secretion of glucose and fructose. Similarly, expression of lactate dehydrogenase- and lactate transporter-encoding genes allowed lactate accumulation in the extracellular medium. Expression of the relevant transporter was essential for secretion. Production of these molecules was further improved by expression of additional heterologous enzymes. Sugars secreted by the engineered cyanobacteria could be used to support Escherichia coli growth in the absence of additional nutrient sources. These results indicate that cyanobacteria can be engineered to produce and secrete high-value hydrophilic products.