?-amylase and ß-galactosidase from the bacteria of Bacillus circulans with the help of Solid State Fermentation method

Because of the gradual development of the enzyme technology, the variety of usage areas and high economic value of products, the researches on the industrial enzyme of bio-technology are becoming more significant.?-Amylase and ß-galactosidase are among the most important enzymes of modern biotechnology. There are a great number of industrial usages of these enzymes. ?-Amylase has many different usage areas such as food, fermenting, textile, paper, the industry of detergent and medicine while ß-galactosidase provides relatively beneficial results for the health and food teachnology by catalyzing the enzymatic hydrolysis of lactose.In our study, we have worked on the obtaining ?-amylase and ß-galactosidase from the bacteria of Bacillus circulans with the help of SSF method and some optimizations of these enzymes have been determined. In order to form SSF medium; cotton pulp, corn gluten meal, the crust of apple, the crust of banana, the crust of orange, rice husk, wheat bran and millet were used as solid substrate. The best result was gained with rice husk. Therefore, rice husk was used as solid substrate in the following studies.The suitable incubation time for the obtaining of the enzymes has been determined as 48th hour and the suitable incubation temperature has been found as 37ºC. In studies done for the purposes of determining the proper planting amount, humidity amount and pH the proper planting amount for ?-amylase has been stated as %25 and for ß-galactosidase as %35 whereas the proper humidity amount has been determined as %20 and suitable initial pH 7.5 for both of the enzymesBy using 50mM NaCI, %1 CHAPS, %1 Triton, %1 SDS, %1 Tween 40, deionize water, pH 6.8 and pH 7.0 sodium, the buffer of phosphate and tap water, the proper environment of extraction was specified. The best result for both enzymes was gained by using the tap water.While designating the activations done at temperatures around 25 ºC ? 90 ºC, the ideal activation temperature for ?- amylase was determined as 70ºC and for ß- galactosidase as 60ºC.While obtaining the enzymes, the best activation was seen with the mixture of 0.5g millet and 4.5g rice husk, out of the other solid substrate blends.The effect of different nitrogen and carbon sources in % 1 ratios on the production of enzymes was observed. While the best nitrogen source for ?-amylase was determined as ammonium chloride and ammonium nitrate it was found that there is no effect of nitrogen sources on ß-galactosidase in terms of increasing the activation. It was determined that carbone sources in % 1 ratios do not induce the activation of enzymes.The effect of metal ions in %0.1 ratios on the activation of enzyme was observed. Enzyme activity could not be seen in ZnSO4.7H2O and CuSO4.5H2O for both enzymes. It was revealed that the amount of MgSO4.7H2O, FeSO4.7H2O and CaCl2 was lower than the control group.Key words : SSF, ?-amylase, ß-galactosidase, Bacillus circulans