Irradiation Of Atp Bioluminescent Method To Detect The Effect Of Bacteria Bacteria Content In The Dried Vegetables

ATP(adenosine triphosphate)bioluminescence assay has convenient and fast characters. At the same time, it has a specificity and high sensitivity to the ATP determination. As a result, the detection of ATP concentration using bioluminescence assay receives a far-ranging application in the germ of clinical medical science and accepted by World Health Organization(WHO). This technique has already more and more used for a bacterial inspection in the developed countries currently now.The theoretics of ATP bioluminescence assay lies in the fact that ATP is widespreadly existed to the whole live cell including bacteria. Under the certain condition, the content of ATP is stablely. ATP will be decreased by the enzyme of itself at the die time of bacteria. The ATP content becomes a basis that the sample have a germ whether. In the method of the content assay of ATP, the determination method of luciferin-luciferase is the most intelligent. At the time of the whole substrate in excess, luminous intensity become a direct proportion with the quantity of ATP. Consequently, it can be simply reckon a result of the bacteria content.However, YueGao, etc.(2007),to detect the bacteria content of dry shallot powder and pepper with the ATP bioluminescence assay by irradiation sterilization. It appeared a phenomenon that the luminous intensity of ATP extract was inconsistent with the actual bacteria content of sample. The reason of effect is still not clear and has never seen to be reported as well currently.To make the assay that detection of ATP concentration using bioluminescence applied in sterilizing by irradiation, the paper carry on a search around the effect that irradiation on the impact to the detection of ATP concentration using bioluminescence assay of the bacteria content to dehydrated vegetables. Results show that:1. ATP bioluminescence assay to examine bacteria content of irradiated dehydrated vegetables’. The result displayed that luminous intensity had inconsistent with actual bacteria content. The irradiation on the impact to the ATP bioluminescence assay of samples had universality2. Results of spectrum analysis showed that:1) Bioluminescence spectrum had the same wavelength range, also no significant difference on peak values of bioluminescence spectrum before-and-after irradiation test on ATP extract of Samples. According to the two spectrum characters, irradiation had not had the influence to the sample’s luminescence quality. This can be identified that the effect that irradiation on the impact to the ATP bioluminescence assay of the bacteria content to dehydrated vegetables was displayed by bioluminescent system of ATP.2) Irradiation on ATP extract of samples had an impact on the peak value of the bioluminescent spectrum. The raise of the peak value of the bioluminescent spectrum showed that irradiation increased the ATP content in extract of samples.3. Irradiation on dehydrated vegetables sterilized by high temperature. Used ATP bioluminescence assay to detect the luminescence intensity of samples., results showed that the luminescence intensity had no significantly different from the CK, indicating that the dehydrated vegetables itself did not affect luminescence intensity of ATP after irradiation; In the application of ATP bioluminescence assay to measure the bacteria content of E. coli bacterium fluid irradiated. It appeared a phenomenon that the luminous intensity of ATP extract was inconsistent with the actual bacteria content of sample. It could be confirmed that bacteria make the effect.4. The luminous intensity of free ATP which extracted from E. coli bacterium fluid irradiated was higher than CK. It can be deduced that the number of free ATP in the sample had increased for the raise of luminous intensity obviously. Radiation treatment could not increase the bioluminescence intensity of ATP from the LB culture media steriled. The ATP increased from E. coli of samples irradiated only.5. Through to the total ATPase activity determination of E. coli, results showed that the greater the radiation did, the lower the total ATPase activity. ATPase is the ATP-specific catabolic enzyme. ATPase activity decreased, so the degradation of ATP from dead bacteria was blocked, thus increasing the content of free ATP in the samples.6. When the free ATP was removed, the bioluminescence intensity of ATP extract was consistent with the actual content of bacteria. This may explained that the influence was due to the increase of content of free ATP from samples.In summary, the ATPase activity of bacterium was reduced when irradiation kill the bacteria of samples. As a result, the ATP content in bacteria didn’t decreased with the killed of bacteria, which contributed to the increase of free ATP in ATP extract and finally led to the disagreement between the bioluminescence intensity of ATP extract was consistent with the actual number of bacteria. In the application of ATP bioluminescence assay to detect the sterilized effect of products by irradiation, it must be consider that free ATP of samples would impact on the detection.This research will expand the application domain of the ATP bioluminescence assay and perfect the certain theoretical and practical significance.