The P170 expression system enhances hyaluronan molecular weight and production in metabolically-engineered Lactococcus lactis |
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Institution: | 1. Department of Biochemistry and Molecular Biology, Federal University of Paraná, CP 19046, CEP 81531-980 Curitiba, PR, Brazil;2. Department of Cell Biology, Federal University of Paraná, CEP 81531-980 Curitiba, PR, Brazil |
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Abstract: | Recombinant Lactococcus lactis strains based on the P170 expression system were developed for hyaluronan (HA) production, by incorporating genes from the has operon of Streptococcus zooepidemicus and compared with nisin-inducible recombinant L. lactis strains containing the hasABC and hasABD constructs. It was found across all batch and fed-batch experimental studies that HA concentration and molecular weight (MW) were higher for the P170 expression systems than the corresponding NICE-based strains. The highest hyaluronan MW was obtained for all constructs in batch studies at 60 g/L initial glucose concentration, the highest being 2.94 MDa for the P170 strains with hasABC construct (L. lactis APJ3). In fed-batch studies with constant feed rate, the L. lactis APJ3 gave better HA yield (0.03 g/g) than the NICE-based strain. A higher hyaluronan MW was obtained for all strains in pulse fed-batch compared to constant feed experiments, the highest being 2.52 MDa for L. lactis APJ3. |
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Keywords: | Hyaluronan Biosynthesis Recombinant DNA Batch Processing Fed-Batch Cultures |
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