|
KMID : 1007519960050010085
|
|
Food Science and Biotechnology
1996 Volume.5 No. 1 p.85 ~ p.89
|
|
|
Clarification of Recombinant E . coli Lysate Using Submicron - Sized Polymeric Particle
|
|
Kim Chan-Wha
Rha Cho-Kyun
|
|
|
Abstract
|
|
|
|
A submicron-sized polymeric particle (SSPP) was utilized to clarify a xylose isomerase overproducing E. coli lysate. The optimum concentration range of positively charged SSPP was 0.10 to 0.15 g SSPP/g wet cell E. coli. In the clarification process, only 27 to 40% of the total xylose isomerase was retained in the supernatant, suggesting significant adsorption of the product enzyme on SSPP. The adsorption of xylose isomerase on SSPP could be controlled by using potassium phosphate. The. concentration of the product enzyme retaining in the supernatant increased with increases in the concentration of potassium phosphate. At the concentration of potassium phosphate above 70 mM, more than 93% of the total enzyme could be retained in the supernatant. The turbidity of the supernatant decreased to less than 0.01 at 600㎚ by standing for 8 hr or centrifugation at 700×g for 5 min. The concentration of DNA in the supernatant decreased to less than 5% of the total DNA in the cell lysate. The concentration of endotoxins decreased to less than 0.3 ㎍/㎖, a reduction of three orders of magnitude. In addition, 40% of the total proteins could be removed as undesired proteins in this clarification process.
|
|
|
KEYWORD
|
|
|
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
 
|