Screening and strain improvement of lysine amino acid Presented by: SadamHussainShaikh
Lysine amino acid • It is an essential amino acid which is not prepared in human body . • Must be supplied by diet . • It has net positive charge. • Commonly found on the surfaces of proteins and enzymes .
SOURECES • Red meat • Fish • Milk • Eggs • Cheese • Vegetables • Microbes • Above all are rich sources of lysine except vegetables.
Screening of lysine Screening is a method to determine particular or individual organism , protein , amino acid so on . A screening test tells us the presence of that amino acid which is needed for outcome of interest. Agricultural by products are used as low cost carbohydrates sources for microbial production of high values added products such as amino acids .
Continued…lysine Can be produced by different methods as given below .1 Chemical synthesis2 Enzymatic method 3 Protoplast fusion technology4 Recombinant DNA technology5 Fermentation technology
Continued… • Among all above methods , fermentation is the most economical and practical means of producing lysine , as in this method low temperature , low pressure and low cost of carbon sources are used and a biological form lysine (L-lysine) is produced. • The selection of organism is very important for this method , so we first select the organism .
Materials and Methods • Microorganism: • C . Glutamicumis a Gram positive organism , most of the strains are able to utilize acetic acid , ethanol , glucose to produce lysine amino acid and also l-glutamate . • Activation and screening: • The bacteria were activated by incubating at 30 degree Celsius for two days in Nutrient broth media . • Then the tri sector cultivation should be performed in plate count agar media in order to isolate the bacteria.
Cultivation and lysine production • The inoculums should be introduced in seed culture and incubated in a rotary shaker at 120 rpm and for 30 Celsius. After 18 hrs , the seed culture pipetted in basic medium with the same condition at 180 rpm . • Lysine accumulation determined by sampling from the broth culture within 5 days. • Selection of the best nitrogen source: • Finding the best nitrogen source for growth of the organism and lysine accumulation , we replace ammonium sulphate of basal medium with ammonium chloride , ammonium dihydrogen phosphate , ammonium acetate and potassium sulphate.
Qualitative Assay The Thin Layer Chromatography technique is technique through we may can detect the L-lysine in culture broth . Quantitative estimation: Quantitative estimation of lysine in the supernatant fluid was determined by the same method mentioned above .