Studies On The Extraction,Purification And Activities Of Peptidoglycan From Actinoplanes Sp. Fermentation Residue

Actinoplanes sp.was commomly used in the production of hypoglycemic agents-Acarbose.Meanwhile,a large amount of fermentation residue was discharged from this process.The traditional treatment of residues was complicated and with high cost.Active components were found with large quantities in the fermentation residue.Therefore,it is of great value to develop an efficient extraction method to obtain active ingredients in the residue.This paper is devoted to the extraction,separation and biological activity evaluation of peptidoglycan from the fermentation residues of Actinoplanes sp.,providing a good prospect for the reusing of bio-pharmaceutical wastes.1)Based on the single factor tests,the response surface methodology(RSM)was used for optimizing mechanochemical extraction processing of peptidoglycan from Actinoplanes sp.The results of the optimum milling conditions were as follows: NaOH(6%),ball to power weight ratio(15),rotation speed(300 rpm),milling time(5 min).The orthogonal experiment was used to optimize the protein-depositing method.The optimum conditions were as follows: rotation speed(300 rpm),SDS(17%),ball to power weight ratio(15),milling time(30 min).Finally,a simple,time-saving and low cost extraction route was obtained.The obtained extract was confirmed to be peptidoglycans by lysozyme dissolution test,infrared spectroscopy and chemical composition analysis and so on.2)The extract obtained in part 1 was used as raw material to obtain purified soluble peptidoglycan component SPG-1 by enzymolysis of lysozyme and semi-preparative liquid phase.Immunization and antitumor activities of the component SPG-1 were studied.The results were as follows: The proliferation of splenic lymphocytes in rats showed that SPG-1 had significant proliferative effects on T and B lymphocytes.In vitro stimulation of macrophages indicated that the SPG-1 could stimulate the production of IL-10,IL-12 and TNF-? by macrophages,and the anti-tumor test results illustrated that SPG-1 has a good inhibitory effect on cancer cells,such as HT29 colon cancer cells,Eca-109 esophageal cancer cells and Hep G2 liver cancer cells.