Hyaluronic acid production is enhanced by the additional co-expression of UDP-glucose pyrophosphorylase in Lactococcus lactis.
Hyaluronic acid (HA) production was metabolically engineered in Lactococcus lactis by introducing the HA synthetic machinery from the has operon of the pathogenic bacterium Streptococcus zooepidemicus. This study shows that the insertion of uridine diphosphate (UDP)-glucose pyrophosphorylase (hasC) gene in addition to the HA synthase (hasA) and UDP-glucose dehydrogenase (hasB) genes has a significant impact on increasing HA production. The recombinant L. lactis NZ9000 strain transformed with the plasmid pSJR2 (co-expressing hasA and hasB genes only) produced a maximum of 107 mg/l HA in static flask experiments with varying initial glucose concentrations, while the corresponding experiments with the transformant SJR3 (co-expressing hasA, hasB, and hasC genes) gave a maximum yield of 234 mg/l HA. The plasmid cloned with the insertion of the full has operon comprising of five different genes (hasA, hasB, hasC, hasD, and hasE) exhibited structural instability. The HA yield was further enhanced in batch bioreactor experiments with controlled pH and aeration, and a maximum of 1.8 g/l HA was produced by the SJR3 culture.
Department of Biotechnology, Indian Institute of Technology Madras, Chennai 600036, India.
Shashi Bala
SB
Prasad
Guhan
G
Jayaraman
K B
KB
Ramachandran
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001709:Biotechnology / Q000379:methods; D015964:Gene Expression Regulation, Bacterial; D005818:Genetic Engineering / Q000379:methods; D006820:Hyaluronic Acid / Q000096:biosynthesis*; D013294:Lactococcus lactis / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D008025:Ligases / Q000235:genetics / Q000378:metabolism; D009876:Operon; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D018502:Streptococcus equi / Q000201:enzymology / Q000235:genetics; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase / Q000235:genetics / Q000378:metabolism*; D014533:Uridine Diphosphate Glucose Dehydrogenase / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D006820:Hyaluronic Acid; D014533:Uridine Diphosphate Glucose Dehydrogenase; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase; D008025:Ligases