Evaluate the effectiveness of aseptic techniques you conducted
Throughout the experiments conducted aseptic techniques played an important role throughout. Aseptic techniques is a procedure that is used to prevent contamination of pathogens. The aseptic techniques I conducted were for the handling of equipment. While conducting the experiment I made sure that windows and doors had been closed. This is so that I draughts are reduced and to avoid the experiment being disturbed by sudden movements, following this procedure did not allow any foreign pathogens to contaminate the experiment. I made sure that the equipment was set up near where I was working and the work tables were disinfected with ethanol before conducting the experiment. This avoided infection of all the equipment used which was important as this prevents incorrect results. This was shown when there were no irregular results that were from the experiment. The equipment that was used was sterilised before and after the exposure such as a glass spreader was placed in a beaker of ethanol to kill the bacteria that may be exposed to it.
Why is it important to find out if an antibiotic is a primary or secondary metabolite before bacteria are grown on a large scale to produce the antibiotic?
It is important to know if an antibiotic is primary or secondary because it helps to find out the best conditions they work good at such as the temperature, PH and water content. Primary metabolites are formed from fermentation; the products include ethanol, aseptic acid and lactic acid. But because of high product yield and low reproducibility cost and major interests were shown within respective markets. Secondary metabolites involve antibiotics, which were first identified and defined as chemical compounds, which were made from a microorganism; these microorganisms have the ability to stop growth and to destroy bacteria and microorganisms in the statue of diluted solutions.
There is always a competitive repetitive search for antibiotics. Penicillin is a type of antibiotic and a secondary metabolite. This antibiotic is made when there is a growth of the fungus and becomes inhibited by stress, but this is not produced during the period of active growth. Penicillin is bactericidal to a lot of gram-positive bacteria, it also acts as an inhibiting transpeptidation and this means that the ability to prevent new cells forming walls.
What practical investigations would you need to do to find the best growth of bacterium which produce the antibiotic?
The conditions has to be determined before finding the best growth of bacterium. The optimum growth conditions of useful microorganisms will occur in laboratory flasks. Within these flasks investigations will occur to determine the optimum nutrient, temperature, oxygen and the varied PH requirements of an organism. To find the growth rates dilution plating, viable cell counts and direct cell counts using a haemocytometer and turbidity will be included. The production of antibiotics this undergoes several stages. Primary metabolism is the first part of the process, the media for this stage is focused on achieving the full growth and biomass production. Once the theoretical aim for the biomass had been achieved the culture is starved and the stress level conditions would be induced which will start the production of the antibiotic. While conducting this method of production the fed batch method is required to feed the culture. This method enables you to add substrates to the reactor in order for small amounts.
The penicillin chrysogenum often contains carbon sources which is usually found within corn,liquor and also glucose. The medium of these are also added to the fermenter, the medium also contains salts and sodium nitrates. They also provide essential ions which are required for fungus metabolic activities
Fermentation is done in a fed batch process where glucose should not be added with large amounts from the start of the growth process. This will result in low yield penicillin production, because a large amount glucose which prevents the penicillin production. From the fermentation process there is specific conditions which are required, (temperature at 20-24 degrees Celsius, also a pH condition at 6.5). Another condition is the pressure with a bioreactor, which must be higher regular atmospheric pressure in order to prevent foreign contaminants from entering. Also through the fermentation process it is required for the culture to be mixed well through the culture medium, this enables fungal cells to be able to handle rotation speed around 200rpm.
Seed culture is a process which was developed within a lab through the addition of penicillin chrysogenum spores to liquid medium. When it is grown to a required standard this will then be inoculated into a fermenter. Then the medium ventilated and stressed. Carbon and nitrogen are added with precursor molecules for the penicillin fed batch style method. The regular restrictions are pH, temperature, stirrer speed and also dissolved oxygen concentration are all served throughout the seed culture method. After this is monitored for 40 hours the penicillin is secreted by the fungus. After a full 7 days the growth process is finished, the pH increases to 8.0 and above and the penicillin production is stopped.