03270ntm a2200289 a 4500 vtls000100062 KUKTEM 20251117113333.0 170504t2017 my da f abm 000 0 eng d THE0000132(Local) 201905131240 nazirah 201710041227 aishah 201705041633 saini UMP FIST .C45 2017 r Thesis Chia, Vi Vien Production of L-asparaginase through biodegradation of chicken bone wastes / Chia Vi Vien Kuantan, Pahang : UMP, 2017 xix, 113 p. : ill. (some col.) ; 30 cm. + 1 CD-ROM Faculty of Industrial Sciences and Technology Thesis (Master of Science (Biotechnology)) -- Universiti Malaysia Pahang – 2017 Bibliography : p. 93-106 L-asparaginase is highly demanded in pharmaceutical, food and biosensor industry due to its remarkable properties in hydrolysing L-asparagine into aspartic acid and ammonia. Owing to this significant property, L-asparaginase is widely produced through microbial fermentation. This research aims to produce L-asparaginase enzyme through microbial fermentation by the most potent isolate in the presence of chicken bone wastes as the substrate. Therefore, this research can be categorised into 3 parts where the first part was to perform several studies in determining the best enzyme producer and substrate while the second part was the effect of process parameters using One-Factor-at-a-Time (OFAT) method and the third part was the purification and characterisation study of L-asparaginase. Six millilitres (5 x 108 cells/ml) of Escherichia coli ATCC 10536 is the optimum inoculum size in 50 ml of pH 9 growth media at 40 °C for 2 days. In addition to that, Escherichia coli ATCC 10536 was fully enhanced when it was engaged with 1.0 % w/v of starch, 0.2 % w/v of ammonium chloride and 1 % w/v of cooked chicken bone (CCB) as the carbon source, nitrogen source and substrate respectively. The effect of process parameters studied using OFAT method revealed that Escherichia coli ATCC 10536 is the most potent L-asparaginase producer in this research and it preferred nutrient broth as the growth media in producing L-asparaginase. Furthermore, Escherichia coli ATCC 10536 L-asparaginase was purified by ammonium sulfate precipitation, dialysis and DEAE-cellulose chromatography. At the end of purification, 0.42 % yield of L-asparaginase was obtained. In addition, the dialysed ammonium sulfate fraction of L-asparaginase was partially characterised which resulted to 40 °C, pH 8, Na+ and ethylenediaminetetraacetic acid (EDTA) being the optimum incubation temperature, pH, metal ion and inhibitor respectively. In conclusion, Escherichia coli ATCC 10536 is an excellent L-asparaginase producer when the fermentation is supplemented with high protein content of cooked chicken bone as substrate. Faculty of Industrial Sciences and Technology Dissertations Universities and Colleges Dissertations Theses http://ecollib.ump.edu.my/25890/ Library access only 0 0 lcc 0 1 10000 10000 2019-09-04 0 FIST .C45 2017 r Thesis 0000117815 2019-09-04 00:00:00 1 2019-09-04 THESIS 0 0 lcc 0 1 10000 10000 2019-09-04 0 CD 10764 | FIST .C45 2017 r Thesis 0000117816 2019-09-04 00:00:00 1 2019-09-04 THESIS VIRTUA40 6983 6989 VTLSSORT0080*0200*0400*0900*1000*2450*2600*3000*5000*5020*5040*5200*6100*6500*6501*8560*9992