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Method for preparing blood sugar reducing peptide by fluorescence spectrum monitoring enzyme method

A fluorescence spectrum and blood sugar lowering technology, applied in fluorescence/phosphorescence, material analysis through optical means, measuring devices, etc., can solve problems such as difficult to stabilize and reliable product quality, achieve great social value, huge economic value, and improve quality Effect

Active Publication Date: 2017-08-01
哈尔滨岿创生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no effective monitoring method in the process of preparing hypoglycemic peptides by enzymatic hydrolysis, which makes the quality of the product difficult to be stable and reliable, which is extremely disproportionate to modern industrial production

Method used

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  • Method for preparing blood sugar reducing peptide by fluorescence spectrum monitoring enzyme method
  • Method for preparing blood sugar reducing peptide by fluorescence spectrum monitoring enzyme method
  • Method for preparing blood sugar reducing peptide by fluorescence spectrum monitoring enzyme method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Take 50mL of 0.8% (W / V) peony seed protein solution, add 5000U / g of trypsin, keep the pH at 8.0, and keep the temperature at 37°C at 0, 3, 6, 9, 12, 15, 20, 30, 45, Take 20 μL of sample at 60 minutes and add 4 mL of PBS buffer solution with pH 8.0 and 0.05 mol / L to dilute.

[0046] Use the F-4600 fluorescence spectrometer to scan the spectrum, the excitation wavelength is 280nm, the emission wavelength is 300-500nm, the wavelength resolution is 0.2nm, the scanning speed is 1200nm / min, the delay time is 0.5s, and the voltage is 700V to obtain the fluorescence spectrum, such as figure 1 shown.

[0047] Calculate the first-order derivative of the obtained fluorescence spectrum, the window width is 21 points, and the wavelength value when the derivative value is 0 is obtained through numerical calculation, which is the peak value of the fluorescence spectrum, as shown in Table 1.

[0048] Table 1 Comparison table of peak position and time between ultrasonic enzymatic hydro...

Embodiment 2

[0052] Concentration 0.8% (W / V) of peony seed protein solution 50mL, add trypsin amount 5000U / g. Using ultrasonic enzymatic hydrolysis method, ultrasonic frequency 28kHz, power 100W, sample 20μL at 0, 3, 6, 9, 12, 15, 20, 30, 45, 60min respectively, add 4mL PBS with pH8.0 concentration 0.05mol / L Buffer dilution.

[0053]Use F-4600 fluorescence spectrometer to scan the fluorescence spectrum. The excitation wavelength is 280nm, the emission wavelength is 300-500nm, the wavelength resolution is 1nm, the scanning speed is 1200nm / min, the delay time is 0.5s, the voltage is 700V, and the fluorescence spectrum is collected. The results are as follows: figure 2 shown.

[0054] Calculate the first-order derivative of the obtained fluorescence spectrum, the window width is 5 points, and the wavelength value when the derivative value is 0 is obtained through numerical calculation, which is the peak value of the fluorescence spectrum, as shown in Table 1.

[0055] The peak position va...

Embodiment 3

[0058] 0.8% (W / V) wheat germ protein solution 50mL, the use of power of 60W, ultrasonic frequency of 65kHz, ultrasonic pretreatment at 48 ℃ for 27min, to achieve the pretreatment of wheat germ protein. Add trypsin for enzymatic hydrolysis, and the amount of enzyme added is 25000U / g. At 0, 2.5, 5, 7.5, 10, 12.5, 15, 17.5, 20, 25, 30, 40, and 50 minutes, 50 μL of samples were taken respectively, and diluted by adding 4 mL of PBS buffer solution with a concentration of 0.05 mol / L at pH 8.0.

[0059] Using F-4600 fluorescence spectrometer for detection, the excitation wavelength is 280nm, the emission wavelength is 300-500nm, the scanning speed is 1200nm / min, the delay time is 0.5s, the emission wavelength interval is 0.2nm, and the voltage is 700V for scanning. Spectral results such as image 3 shown.

[0060] Calculate the derivative of the obtained fluorescence spectrum, the window width is 11 points, and the wavelength value when the derivative is 0 is obtained through numer...

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Abstract

The invention discloses a method for preparing blood sugar reducing peptide by a fluorescence spectrum monitoring enzyme method. The method comprises the following steps: (1) collecting fluorescence spectra of enzymolysis reaction liquid at different time; (2) acquiring peak positions of the fluorescence spectra; (3) establishing relation between the peak positions of the fluorescence spectra and an inhibiting rate of enzymolysis protein fluid to the activity of alpha glucosaccharase; (4) monitoring the inhibiting rate of the enzymolysis protein fluid to the alpha glucosaccharase in real time. According to the method, the fluorescence spectra are collected during the preparation process of the blood sugar reducing peptide, and are subjected to analytical calculation, thus the inhibiting rate of an enzymolysis reaction product to the activity of the alpha glucosaccharase can be obtained, and the advantages of convenience and quick speed are achieved; besides, a chemical reagent is not needed in a detection process, the detection cost is low, environment pollution is not caused, and compared with a wet chemical detection method, these advantages are incomparable; meanwhile, the quality of the blood sugar reducing peptide produced in an industrialized way can be improved, and therefore the product can better reduce blood sugar concentration of a glycosuria patient, thus improving the health level of people, and having great social value and huge economic value.

Description

technical field [0001] The invention belongs to the field of food biotechnology, and in particular relates to a method for producing hypoglycemic peptides by using fluorescence spectrum monitoring enzymatic method. Background technique [0002] With the continuous improvement of the economic level, people's dietary conditions are getting better and better. However, lack of exercise and unreasonable daily routines lead to an increase in the incidence of diabetes year by year. Long-term hyperglycemia can lead to complications such as proteinuria, hypertension, atherosclerosis, nervous system lesions, and infections, which seriously affect human health. [0003] α-glucosidase inhibitors are a class of substances that inhibit the activity of α-glucosidase in the small intestine, which can significantly reduce people's postprandial blood sugar. The development of new α-glucosidase inhibitors is the direction of efforts of many researchers. In recent years, there have been repor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 颜辉余永建朱胜虎贾俊强熊孟江明珠吴琼英
Owner 哈尔滨岿创生物科技有限公司
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