MICROBIAL PRODUCTION OF BIOSURFACTANT FROM LOCAL SOURCES
Biosurfactants are structurally diverse group of surface active substances produced by microorganisms. The microorganisms that produce Biosurfactants include Pseudomonas, Bacillus, Micrococcus, Mycobacterium, Rhodococcus etc. All Biosurfactants are amphiphiles: consists of two parts –a polar (hydrophilic) moiety and non-polar (hydrophobic) group. They have ability to break complex lipid to smaller units. They reduce surface tension of different industrial and natural media. They have ability to stabilize emulsions in various industrial applications. The advantages of Biosurfactant over chemical surfactants include; pH tolerant, less toxic to the environment, biodegradable, better foaming properties than surfactants, and biosurfactant can be produced from agro based industrial wastes. The applications of Biosurfactant are wide but are not limited to: Food Industries, Agriculture, Cosmetic Industries and House hold, pharmaceutical Industries, Oil and Gas Industries. The production of Biosurfactants in Nigeria from local sources saves a lot of foreign exchange earnings is a major justification for this project. The project is expected to bring collaboration between FIIRO and major upstream, Pharmaceutical industries e.t.c in Nigeria The product is used for most of our mandates (Food, Environmental technology etc). The Institute through this research is expected to solve the oil spill problems in the Niger Delta through the use Biosurfactants to degrade recalcitrant fractions of Petroleum hydrocarbon (e.g. Ogoni Spill). The aim of this project is to Develop biotechnological techniques for the production of biosurfactant for small, medium and large scales industrialization in Nigeria. The objectives are to: To screen and isolate biosurfactant producing microorganisms, characterise and identify biosurfactant producing microorganisms using traditional microbiology and molecular biology tools, produce biosurfactant in submerged fermentation method, determine the optimum cultural conditions for biosurfactant production, characterise the produced Biosurfactants by determining its pH tolerance, thermal stability, antimicrobial properties on food spoilage organisms et, apply the produced biosurfactant into the production of cosmetics, shampoos, detergents, and degradation of petroleum hydrocarbon polluted Agricultural soil, carry toxicity studies of produced biosurfactant on Animal model. For the isolation of biosurfactant producing bacteria, soil samples were collected from crude oil polluted soil undergoing bioremediation in Port Harcourt. The soil samples were collected with the use of soil auger into sterile polythene and stored at low temperature till use. The isolated colonies were tested for their Biosurfactants production by three methods: Oil Spreading Technique, Blood haemolysis test, Emulsification stability test, Identification of biosurfactant hyper-producing strains, Molecular biology method was employed in the identification, DNA isolation was done using Inqaba (SA) universal isolation kits (used according to manufacturer’s). The PCR mixture comprises of; 1μl of 10X buffer, 0.4μl of 50mM MgCl2,0.5μl of 2.5mm dNTPs, 0.5μl 5mM forward primer, 0.5μlof 5mM Reverse primer, 0.05μl of 5units/μlTaq with 2μl of template DNA, 2ul of 5x sequencing buffer and 1μl of 50% DMSO and 5.05μl of distilled water to make-up 10μl reaction mixture. Initial denaturation temperature of 94oC for 3minutes, 30 cycles of 94oC for 60seconds, 56oC for 60seconds, and 72oC for 120seconds were employed. For final extension a temperature of 72oC for 10 minutes was employed. Blasting was done using the nucleotide BLAST software of the National centre for Biotechnology Information (NCBI). The Research group is currently carrying some repeat experiments in the area of screening and optimization using various local sources of carbon and nitrogen. The research group is also investigating the biosurfactant producing potentials of yeasts, cyanobacteria and algae from crude oil polluted environments in Nigeria. Thereafter applications in various areas of need will commence. Research grants, collaborations, staff exchange etc are hereby solicited for.