The Study On The Non-enzymic Hydrolysis Of Peptidoglycan Support Of Bacterial Spore's Cortex Under High Pressure

This paper studies the HPTS(High Pressure Thermal Sterilization,HPLC,hot auxiliary superhigh Pressure Sterilization)under the condition of spore cortex peptidoglycan(peptidoglycan,PG)hydrolysis change trend,and its hydrolysis mechanism was discussed preliminarily.Chemical bond rupture with PG structure for the study of the main line,first enzyme hydrolysis PG,using different combinations of high pressure,heat and chemical means,through the ultraviolet absorption spectra,reverse phase high performance liquid chromatography(HPLC),matrix assisted laser analytical time of flight mass spectrometry,electrospray liquid-quadrupole ms method and ion trap mass spectrometry analysis of the change of its structure,mass spectrometry technology research under the condition of PG scanning structure and the positive and negative ions fragment ions and the law of cracking is the main feature of PG.Research to the main conclusions are as follows:1?First of all,to study the extraction method of PG,By experiment,the optimal RP-HPLC conditions for analyzing spore's cortical peptidoglycan hydrolysate were as follows:on Welch Ultimate AQ-C18 chromatographic column,column temperature 30 ?,flow speed 1 mL/min,detection wavelength 206 nm,elution time 120 min,with methanol and 0.05 mol/L PBS(pH = 4.31)as mobile phase,in 0-5 min 100%PBS was used,in 5 min-120 min the methanol concentration increased gradually from 0 to 30%.The result indicated that it was feasible to use RP-HPLC for analyzing spore's cortical peptidoglycan hydrolysate,which provided research method support for studying the hydrolysis principle of spore's cortical peptidoglycan under HPTS(High Pressure Thermal Sterilization)treatment,and further for clarifying the principle of spore inactivation by HPTS.2?The rearch of temperature under high pressure and high pressure synergy ethanol was studied in PG.Set different gradient pressure to deal with PG,it was found that:temperature 150 mpa and 300 mpa and 450 mpa with PG,did not find its hydrolysis.450 mpa pressure synergy were 15%,75%ethanol to deal with PG,did not find its hydrolysis.Studied the HPTS processing of PG and inactivated effect and mechanism of bacillus subtilis spores.It was found that:60 ?,530 mpa when handling the 15 min,hydrolysis of PG.This condition synergy pH(4),75%alcohol did not find PG hydrolysis occurs,its have obvious protective effects on PG.60 ?,530 mpa when handling the 15 min,bacillus subtilis spores on clear liquid deposition are significantly higher than not working pressure conditions.3?Study the mechanism of PG hydrolysate auxiliary laser analytical time of flight mass spectrometry(MALDI-TOF)and high performance liquid chromatography(HPLC)-electrospray-triple quadrupole(UPLC-MS/MS)analysis the best conditions.MALDI-TOF mass spectrum scanning level in substance and PG target molecular weight are basically identical.Through to the sample of the goods and the child mother ions scanning method to determine the appropriate qualitative quantitative ion,established the UPLC/MS/MS detection method,a quantitative accuracy,short analysis time,high sensitivity and precision.Qualitative and quantitative detection on subsequent research PG hydrolysate provides methods to support.