Enhancement of recombinant streptokinase production in Lactococcus lactis by suppression of acid tolerance response.
Lactococcus lactis is a potential host for production of recombinant proteins, especially of therapeutic importance. However, in glucose-grown cultures, lowering of pH due to accumulation of lactic acid and the concomitant induction of acid tolerance response (ATR) may affect the recombinant protein produced. In this work, we have analyzed the effect of culture pH and the associated ATR on production of recombinant streptokinase. Streptokinase gene was cloned and expressed as a secretory protein in L. lactis under the control of P170 promoter. It was found to undergo degradation to form inactive products leading to low productivity. The extent of degradation and productivity of streptokinase was greatly influenced by the development of ATR, which was dependent on the pH of the culture and initial phosphate concentration of the medium. It was found that high pH and high initial phosphate concentration leads to suppression of ATR and this results in at least 2.5-fold increase in streptokinase productivity and significant decrease in degradation of streptokinase.
Sriraman
Kalpana
K
Department of Biotechnology, Indian Institute of Technology-Madras, Chennai 600036, India.
Jayaraman
Guhan
G
D000222:Adaptation, Physiological; D001426:Bacterial Proteins / Q000032:analysis; D003001:Cloning, Molecular; D003470:Culture Media / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D013294:Lactococcus lactis / Q000235:genetics* / Q000502:physiology*; D010710:Phosphates; D021381:Protein Transport; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013300:Streptokinase / Q000096:biosynthesis* / Q000235:genetics*
D001426:Bacterial Proteins; D003470:Culture Media; D010710:Phosphates; D011994:Recombinant Proteins; D013300:Streptokinase