Method for determining the activity of phosphoglucose isomerase in plum fruit by spectrophotometer
A technology of dextrose phosphate and spectrophotometer, applied in the field of analytical chemistry, can solve the problems of less research, and achieve the effect of fast analysis speed, good stability and good reproducibility
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Embodiment 1
[0021] The activity of phosphoglucose isomerase (GPI) in different growth and development stages of Furong plum fruit was determined by spectrophotometer, including the following steps:
[0022] (1) Determination of control CK: sucrose invertase converts sucrose into glucose and fructose, and the dehydrogenation of glucose by 6-phosphate glucose dehydrogenase causes the change of absorbance value Abs1=(A1CK-A0CK);
[0023] (2) Preparation of sample extract: take 0.15g of frozen plum pulp sample, grind it with 1.6ml of extract until homogenized; the extract is composed of 50mM Hepes–KOH buffer with pH 7.5, 2mM EDTA, 10mM MgCl 2 , 10mM Dithiothreitol (DTT), 1% (w / v) Triton X-100, 30% (w / v) Glycerol, 1% (w / v) Bovine Serum Albumin (BSA), 5% (w / v) polyvinylpyrrolidone (PVPP), and 1 mM phenylmethanesulfonyl fluoride (PMSF);
[0024] (3) Determination of phosphoglucose isomerase GPI activity in the sample: After the extraction is completed, centrifuge at 12000g for 20min at 4°C, tak...
Embodiment 2
[0027] Embodiment 2 Methodological investigation
[0028] (1) Precision test
[0029] The standard GPI (Sigma) was prepared to a concentration of 0.5 U / μl, and measured 6 times according to the above method, and the relative standard deviation (RSD) was calculated. The test results are shown in Table 1. The results show that the test results are accurate, the instrument precision is high, and the RSD is controlled at about 1.03%.
[0030] Table 1 Precision test
[0031]
[0032] (2) Repeatability test
[0033] Extract the fruit of Hibiscus plum in different periods, extract the enzyme liquid in the fruit according to the requirements of the above step 1, and measure the enzyme activity according to the method of step 3, the results are shown in Table 3, the relative standard deviation RSD of each period is controlled at 2.08%-8.25 %, indicating that the test was repeated well.
[0034] Table 3 Determination of GPI in different growth and development stages of Furong plu...
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