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Determination method of ferulic acid antioxidant activity in vitro

A ferulic acid and production method technology, applied in the field of bioengineering, can solve the problems of cumbersome experimental process, accelerated development of biological products, and long time consumption

Inactive Publication Date: 2014-03-19
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] As an antioxidant, at present, the determination of the antioxidant properties of ferulic acid is gradually being concerned by many researchers, such as through many in vivo experiments, but these experiments are cumbersome and time-consuming. With the development of biological sciences With continuous expansion, it will be more convenient and faster to use biotechnology means combined with simple chemical methods to measure some properties, which will accelerate the development of biological products and biotechnology in more fields

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  • Determination method of ferulic acid antioxidant activity in vitro
  • Determination method of ferulic acid antioxidant activity in vitro

Examples

Experimental program
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Embodiment 1

[0020] The preparation of embodiment 1 ferulic acid

[0021] Using pretreated wheat bran as substrate, weigh 0.5 g into a shake flask, add 3.72 mL of co-expressed crude enzyme solution, and then add phosphate buffer (1 mM, pH 5.0) to a final volume of 30 mL. The reaction system was kept at 40 °C for 12 h, the enzymatic hydrolyzed solution was collected, and 10 mL of bran hydrolyzed solution was dynamically adsorbed on a macroporous resin column at 1 mL / min. After the adsorption was completed, it was allowed to stand for 2 h, and then slowly eluted with distilled water with a flow rate of 1 mL. / min, collect the eluent in batches, after the water washing solution no longer contains impurities such as reducing sugar and pigment, use 50% ethanol for elution, the flow rate is 1.5mL / min, collect the eluent, put it in a vacuum It was concentrated in a rotary evaporator to a final volume of 10 mL, and the content of ferulic acid was detected by HPLC to be 1.75 mg.

Embodiment 2

[0022] Example 2 In vitro antioxidant activity verification of ferulic acid

[0023] (1) Determination of reducing power

[0024] Add 1 mL of 0.2M PBS buffer (pH 6.6), 2 mL of 1% potassium ferricyanide solution and 1 mL of ferulic acid sample solutions of different concentrations to a stoppered test tube, react in a constant temperature water bath at 50°C for 20 min, cool immediately, add 1.0 mL of 10 % trichloroacetic acid solution, centrifuge at 4000 rpm for 8 min, take 2 mL of supernatant, add 1 mL of 1% ferric chloride solution and 2.0 mL of distilled water, shake and mix, after standing for 5 min, measure the absorbance values ​​at 700 nm respectively, the result of ferulic acid The reducing ability of ferulic acid showed a steady growth trend with the increase of concentration, and showed a certain dose-effect relationship. The reducing ability of 20 μg / mL ferulic acid was 5.29 times that of 4 μg / mL ferulic acid. As a control, distilled water was used instead of ferulic...

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Abstract

The invention aims at providing a determination method of antioxidant activity in vitro of enzymatic hydrolysate ferulic acid of co-expressed feruloyl esterase / xylanase. The ferulic acid is obtained by hydrolyzing wheat bran by using the co-expressed enzyme solution produced through a yeast system (GS115 / faeA-xyn11A) of the co-expressed feruloyl esterase A and xylanase, and the antioxidant activity in vitro of the ferulic acid is determined from three aspects of reducing power, DPPH free radical and hydroxyl free radical scavenging rate. The experimental result proves that the ferulic acid has good reducing capacity and free radical scavenging capacity. The determination method is simple and effective, convenient and fast, and has a certain potential value on the antioxidant determination aspect of the ferulic acid in industrial application.

Description

technical field [0001] The invention relates to the in vitro antioxidant activity measurement of ferulic acid, an enzymatic hydrolysis product of co-expressing ferulic acid esterase / xylanase, from three aspects: reducing power, DPPH free radical and hydroxyl radical scavenging rate, and belongs to bioengineering technology. field. Background technique [0002] Ferulic acid esterase (EC3.1.1.73) can catalyze the hydrolysis of ester bonds between ferulic acid and its derivatives and polysaccharides in plant cell walls, and release ferulic acid therefrom. In recent years, ferulic acid esterase has been widely used in feed, paper, food, textile, medicine, bioenergy and other fields, so researchers have pushed the development and utilization of ferulic acid esterase to a new peak. [0003] Ferulic acid is the most abundant hydroxycinnamic acid, which is the main medicinal component of Angelica sinensis and Chuanxiong. Ferulic acid is an effective natural antioxidant with high s...

Claims

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Application Information

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IPC IPC(8): C12P7/42G01N21/31
Inventor 邬敏辰殷欣张鹏龚燕燕李剑芳庞庆丰
Owner JIANGNAN UNIV
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