Bromelain enzyme powder is a protein hydrolyzing enzyme system present in pineapple hanging plants. It has a wide range of uses in the food and pharmaceutical industries. However, its poor stability affects its application, especially in the pharmaceutical industry. In this paper, the effects of humidity conditions temperature conditions and "Coy radiation on pineapple protease were studied with the raw material of pharmaceutical pineapple protease.
1 Materials and methods
1.1 Bromelain raw materials
Bromelain enzyme powder
1.2 Effect of humidity environment
The weighed and dispensed enzyme was placed openly in a closed desiccator (room temperature). Silica gel, NaQ saturated solution and KNO,saturated solution were placed inside the desiccator. The relative humidity of the internal environment was 0.1% (75±1)%2.92%, and samples were taken at regular intervals to determine the enzyme activity.
1.3 Effect of temperature
The weighed and portioned enzymes were placed at temperatures of 1~4°C, 29°C and 55°C respectively, and samples were taken periodically for enzyme activity determination.
1.4 Radiation treatment
The weighed and dispensed enzymes were hermetically sealed in plastic (PVC) centrifuge tubes. The samples were irradiated with "Coy radiation" at room temperature (~20°C). The dose rate was 198 kGy/h and the absorbed doses were 4 kGy8 kGy and 12 kGy. 3 days after irradiation of the samples, the enzyme activity and the total number of bacteria were determined.
1.5 Determination method
Bromelain was determined by FIP method. Tyrosine hydrolyzed from casein was quantified by UV spectrophotometer and the enzyme activity was estimated. Casein was employed as the substrate and hydrolyzed by pineapple protease.
The total number of bacteria was determined using plate colony counting method.
2 Results
2.1 Effect of humidity conditions on the storage of bromelain enzyme powder
The results of the effect of humidity conditions on the storage of bromelain are shown in Table 1.As seen in Table 1, bromelain enzyme activity was relatively stable in the dry environment. The rate of enzyme inactivation was accelerated with the increase of ambient humidity. At the late stage of the experiment, the enzyme activity decreased significantly under high humidity conditions. Only 40.4% 14.4% of the enzyme activity remained after 105 d and 315 d of storage of bromelain, respectively. Whereas, the preservation of enzyme activity was 80.3% and 80.0% under relative humidity (75+1)%.
It can be visualized from the accompanying figure that under dry or low humidity . The inactivation of pineapple protease occurred in the early to late stages of preservation leveled off. This is due to the fact that the humidity of the enzyme is greater than the ambient humidity in the early stage of preservation. The higher moisture imparts a certain flexibility to the enzyme structure. The conformation of the active center of the enzyme is prone to change. When the enzyme reaches the level street with the dry environment, the moisture of the enzyme decreases and the enzyme activity tends to stabilize. It can also be seen from the figure that the enzyme activity has been decreasing under high moisture environment. The reason for this is due to the self-hydrolysis of the protease, in addition to the high moisture increasing the flexibility of the enzyme structure.
2.2 The effect of temperature conditions on the storage of vegetable bromelain enzyme powder
Table 2 displays the findings of the investigation into how temperature affects pineapple protease storage.Pineapple proteases were inactivated at both high and low temperatures, as shown in Table 2, with high temperatures being less stable. Their inactivation rate was faster in the early stage of storage and more moderate in the later stage. The preservation rate of the enzyme after 285 d at 51°C (68.0%) was greater than that after 105 d at high humidity (40.4%). In this case, the effect of temperature on the enzyme was much less than that of humidity. The effect of temperature on the enzyme was much less than that of humidity.
2.3 Effect of 60Cor radiation on pineapple protease activity and bactericidal effect
The results of the effect of 60Cor radiation on pineapple protease activity and sterilizing effect are shown in Table 3. As seen in Table 3, the loss of enzyme activity was 10.6%, 11.0%, and 15.5% when the absorbed dose was 4 kGy 8 kGy, and 12 kGy, respectively. Their sterilization effects were comparable at 99.9%, 100%, and 100%, respectively. In order to minimize the loss of enzyme activity, a dose of 4 kGy or 8 kGy can be selected according to the carryover of the enzyme source. As the amount of fungicide, both sterilization, and reduce the loss of enzyme activity
3 Discussion
In this experiment, the sensitivity of the enzyme to humidity was higher than that to temperature. The enzyme survival rate under silica gel drying environment was better than that under low temperature, and the enzyme inactivation rate under high humidity environment was significantly more than that under 55℃. The high humidity environment increased the moisture content of the enzyme and became the main factor of enzyme inactivation. This is closely related to the self-hydrolysis of pineapple protease. Therefore, in the production, we should pay special attention to the humidity of the environment where the enzyme is stored and the dryness of the enzyme itself. In addition, there is enzyme inactivation when "Coy radiation is used to sterilize pineapple protease. How to further improve the stability of preservation and sterilization of bromelain? sterilization stability of pineapple protease. Further research is needed to select efficient additives, which would be the best way to improve the stability of preservation and sterilization of bromelain enzyme powder. additives will be one of the means.
Guanjie provides bulk bromelain enzyme powder, using the GDU test method. Our production process operates under CGMP standard workshops, utilizing three production lines across two factories, and two independent laboratories. For any inquiries regarding our products, please contact us at info@gybiotech.com.
