Hi Mehmet,
Could you be more specific about what you are looking for in this organism so we can help you in the best way possible?
You may search in database of traditional chinese medicine, and the website is http://tcm.cmu.edu.tw/. The database is the biggest database for drug screening in chinese medicine. There are about 352 kinds of chinese medicine, about 37170 molecules indexed in this database. Wish you for your research.
Hello M. The species of Ganoderma lucidun are well known for their ability to break down fibrous substrates, so if you want to determine a component that differentiates them you can determine their cellulolytic activity by measuring the activity of endo and exoglucanases, B glycosidases, and the presence of xilanolytic enzymes, in the other hand if you want to measure the ligninolityc potential you can look for some others enzyme like polyphenol oxidases such as laccases and peroxidases, if this is what you mean, then I can suggest you some of the most used methods to determine each of them
Regards !!
Hello again I think what you are interested in is knowing about the lignocellulolytic potential of this mushroom strain, you can first determine the cellulolytic capacity by determining the three main enzymes of the cellulolytic complex, which are the b1,4 endoglucanase, the b1,4 exoglucanase , and b-glucosidase. Within these, the main enzyme that begins hydrolysis in the crystalline zone of cellulose are the exoglucanases, these hydrolyze the terminal glycosidic bonds, releasing cellobiose as the main product. They represent 40-70% of the total cellulase system. You can determine the exoglucanases by the method proposed by Adney et al. 2009. Here I leave it explained so you can reproduce it Test with filter paper , Enzymatic activity is determined on enzymatic crudes using the filter paper assay described by Adney. This consists of preparing different test tubes: mixtures of enzymes (enzymatic crude), control of enzymes and target; in the case of the tubes that are going to contain the enzyme mixture, a rolled strip of Whatman 1 filter paper (1.0 x 6.0 cm) equivalent to 50 mg of glucose was placed, which serves as a substrate. Then 1 mL of 0.05 mol / L sodium citrate buffer at pH = 4.8 was added to all the test tubes, except the blank, which was added with 1.5 mL of said solution. The test mixture mixture of enzymes and the blank was then placed in the thermostatic bath at 50 ° C for one minute to balance the substrate with the buffer. Once the system was stabilized, 0.5 mL of the different enzymes were added to the respective test tubes. Then they were placed in the hot water bath at 50˚C for 60 minutes. To stop the reaction and develop color to all, 3 mL of the dinitro salicylic acid (DNS) reagent was added (it develops a color intensity proportional to the concentration of reducing sugars) and for 5 minutes they remained in the heating bath with water at 100˚C. After all the tubes were cooled in a water bath to allow any solid to settle and the absorbance was read at 540 nm in the spectrophotometer, for which a tube was added to each tube. amount of water equivalent to 2.5 mL in 0.2 mL of reaction mixture. A glucose standard curve in 0.05 mol / L sodium citrate buffer and pH = 4.8 was constructed from different standard glucose concentrations (10 mg / mL) and the corresponding absorbance values to determine the concentration of glucose. To determine the enzyme concentration the IUPAC (2008) defines that the formation of 2 mg of glucose / 0.5mL of buffer from 50 mg of Whatman1 paper equals 0.37 IU / mL, therefore by a proportionality relationship Using the above information and determining the glucose concentration through the standard curve, the enzymatic activity of each sample can be determined. Determination of the enzyme exo β1-4 glucanase expressed in international units per milliliter (IU / ml). Which refers to the micromoles of glucose released per minute of reaction under the conditions of the activity test .
Hope this can help you
Regards!
- Badges
- Science topic
- Similar topics
- Biological Science
- Bioscience
- Biotechnology