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The present study is focused upon improving biomass of Kluyveromyces lactis cells expressing recombinant human interferon gamma (hIFN-γ), with the aim of augmenting hIFN-γ concentration using statistical and artificial intelligence approach. Optimization of medium components viz., lactose, yeast extract, and trace elements were performed with Box-Behnken design (BBD) and artificial neural network linked genetic algorithm (ANN-GA) for maximizing biomass of recombinant K. lactis (objective function). The studies resulted over 1.5-fold improvement in the biomass concentration in a medium composed of 80 g/L lactose, 10.353 g/L yeast extract, and 15 mL/L trace elements as compared with initial biomass value. In the same study hIFN-γ concentration reached 881 µg/L which was 2.28-fold higher as compared with initial hIFN-γ concentration obtained in unoptimized medium. Further the batch fermentation study displayed mixed growth associated kinetics with the maximum hIFN-γ production rate of 1.1 mg/L. BBD and ANN-GA, both optimization techniques predicted a higher lactose concentration was clearly beneficial for augmenting K. lactis biomass which in turn increased hIFN-γ concentration.

Biosimilar/Biotherapeutic production is becoming a major area of focus for a big chunk of Biotechnology Industry. Easy licensing and already approved status for clinical use has given it a boost. In the present study, recombinant human interferon gamma was expressed for the first time in Kluyveromyces Lactis (K. lactis) expression system and its expression was optimized by varying growth parameters and carbon source concentration with the aim of increasing recombinant protein production level. Human Interferon gamma gene (IFNG) was cloned in the genomic DNA of K. lactis by homologous recombination and under un-optimized conditions in shake flask, IFN�� protein was secreted in the fermentation medium at a level of 175 ��g/litre quantified by ELISA assay. After optimization of expression conditions using one variable at a time (OVAT) technique, expression level was enhanced by 2.2 folds. Substrate inhibition studies revealed that up to 80 g/l of lactose is well tolerated by K. lactis cells for its growth but more than 80 g/l of lactose causes remarkable reduction in biomass production.

In the present investigation, we have shown a single step purification of IFN-γ from the fermentation broth of recombinant Kluyveromyces lactis (K. lactis) strain, using cationic surfactant based r...

Background: The present study aimed at evaluating the effects of azidothymidine (AZT) on hematologic and biochemical parameters in Charles Foster rats.Methods: Twelve adult healthy Charles Foster rats comprising of six male and six females were selected for study. Test rodents were divided into four groups containing three rodents each. Three males and three females served as control and remaining received AZT drug. Rodents were acclimatized for 10 days and drug was administered for 28 days. After the completion of drug administration, blood samples were collected and analyzed for hematologic parameters, i.e., Hemoglobin (Hb), Packed cell volume (PCV), red blood cell (RBC), Mean corpuscular hemoglobin concentration (MCHC), total leukocyte count (TLC), Mean corpuscular volume (MCV), Platelet count (Plt) using a Fully Automatic Fully Digital Hematology Cell Counter. In addition, biochemical parameters, were measured to assess the effects of AZT on rodent physiology. In-vivo histopathological studies were also performed on vital organs of rodents to understand the effects of drug at tissue level.Results: When compared with the control group, the data indicated a outstanding decrease in Hb, PCV, RBC, TLC and platelets in all test groups, whereas MCHC did not show any major reduction but MCV data suggested a slight increase. Among biochemical parameters, aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP). were found to be remarkably elevated along with elevated bilirubin and reduced albumin, pointing towards a possible liver damage which was later corroborated by liver histopathological study.Conclusions: Above results indicate azidothymidine to be a myelosuppresive and hepatotoxic drug and its usage as an anti-retro viral during highly active anti-retro viral therapy (HAART) regime should be strictly monitored.