KOSEN 한인과학기술자네트워크

우선 프로토콜을 모두 올려보면.. V. FiLTER PAPER ASSAY FOR SACCHARIFYING CELLULASE (FPU Assay) (ref. 8) Substrate: Whatman No. 1 filter paper strip, 1.0 x 6.0 cm (= 50mg). Method 1. Add 1.0 ml 0.05 M Na-citrate, pH 4.8, to a test tube of volume at least 25 ml. 2. Add 0.5 ml enzyme, diluted in citrate buffer. At least two dilutions must be made of each enzyme sample investigated. One dilution should release slightly more and one slightly less than 2.0 mg (absolute amount) of glucose (= reducing sugars as glucose) in the reaction conditions. 3. Temperate to 50°C, add one filter paper strip, mix (NB! it does not matter if a small part of the paper is above the liquid surface, but if the paper “winds“ up the tube it must be pushed down again). 4. Incubate 50°C, 60 mm. 5. Add 3.0 ml DNS, mix. Transfer tube to a rack on the table. 6. Boil for exactly 5.0 mm in a vigorously boiling water bath containing sufficient water. All samples, enzyme blanks, glucose standards and the spectro zero should be boiled together. After boiling, transfer to a cold water bath. 7. Add 20 ml deionized or distilled water. Mix by completely inverting the tube several times so that the solution separates from the bottom of the tube at each inversion (NB. This is important!). 8. When the 'pulp' has settled well, i.e., after at least 20 mm, the color formed is measured against the spectro zero at 540 nm. If the paper pulp does not settle, it will do so after stirring with a glass rod. (The necessity for stirring can be seen after only a few minutes of settling time). 위의 프로토콜이구요.. 추가적으로 당 분석을 하려는데 DNS를 쓰지 않고 당 분석기를 이용하여 glucose를 직접 분석을 하려는데 논문에 나와있는 FPU 계산법을 그대로 따라가도 되는 것인지 궁금합니다. 보통 다른 논문을 보면 셀룰레이즈의 활성이 ?? FPU/g glucan이다..라고 설명을 하곤 하는데 이것이 어떻게 계산이 되어 나오는건지...궁금합니다..