Journal of Biotechnology & Biomaterials
Open Access

Abstract

The production of recombinant proteins in the fermentation process in biotechnology industry can be performed using genetically modified organism derived from Escherichia coli K12. The main health and safety concern on the use of GMOs (Genetically Modified Organism) is related to the potential of the gene transfers to other organisms, toxicity, inducing resistance to antibiotics and compromise biodiversity. The accidental release of GMOs in the recent years has increased the concern with safety and the need for methods able to detect them. Thus, policies of monitoring for disposal to the environment of bacterial carrying plasmids or other genetically modified organisms are required. In our research we associate the methods of membrane filtration and polymerase chain reaction (PCR) as tools for detection and quantification of E. coli K12 in downstream stages and in the final product of a fermentation process. Through contamination and recovery experiments it was possible to determine the detection limit for cells in CFU/mL following the membrane filtration method (1 CFU/mL). PCR reactions performed using E. coli presumptive identification colonies have proved that it is possible to detect E. coli K12 marker genes. Efforts have been undertaken to standardize and validate molecular methodologies in our laboratory among which PCR and more recently real-time PCR.

Biography

Janine Aparecida Correia Duraes Gandra has graduated in Biology and has attended the Master’s degree in Industrial Biotechnology at State University of Montes Claros. As Biologist, she has worked on the Microbiological Quality Control at Novo Nordisk Pharmaceutical Production in Brazil for over 10 years focusing on the following topics: Equipment qualification, validation of analytical methods, preparation of standard operational procedures, training in analytical methods, identification of microorganisms using traditional and rapid methods, sterilization processes. She is a Member of the Validation and Change Control Committee. Currently, she is working in ALP (Achromobacter lyticus protease) production.

Email: jdur@novonordisk.com