CN107373568B

CN107373568B - Method for increasing α -tocopherol content in peanuts

Info

Publication number: CN107373568B
Application number: CN201710662748.1A
Authority: CN
Inventors: 刘国琴; 李淑莹
Original assignee: South China University of Technology SCUT
Current assignee: South China University of Technology SCUT
Priority date: 2017-08-04
Filing date: 2017-08-04
Publication date: 2020-04-28
Anticipated expiration: 2037-08-04
Also published as: CN107373568A

Abstract

The invention discloses a method for increasing the content of α-tocopherol in peanuts. The steps of the method are as follows: washing peanut seeds with water, draining them, soaking them in 0.5-1.5wt% sodium hypochlorite solution for 5-15 minutes for disinfection, rinsing with water until the pH value is neutral, adding water with 2-4 times the mass of peanuts, and placing them in a Soak for 4~12h at 20~30℃ in the dark; germinate for 7 days at 26~32℃ and 70~100% humidity in the dark, spray water every 1~3 hours, and each spray lasts 1~2 minute. The peanut sprouts are vacuum freeze-dried so that the moisture content is 2-3 wt%, pulverized and passed through a 40-60 mesh sieve, vacuum-packed and stored in a drying box for measurement. The method of the invention is simple, low in cost, popular, and can be popularized and applied; the effect is remarkable, and the α-tocopherol content of the treated peanuts is above 178.63 μg/g DW, which is increased by at least 4.87 times, and has the versatility of varieties.

Description

Method for increasing α -tocopherol content in peanuts

Technical Field

The invention relates to the field of improving α -tocopherol content in plants, in particular to a method for improving α -tocopherol content in peanuts.

Background

Vitamin E (Vitamin E), also known as tocopherol (tocophenol), is a naturally occurring fat-soluble Vitamin, including both tocopherols and tocotrienols, 8 are classified into α -, β -, delta-, gamma-tocopherols and α -, β -, delta-, gamma-tocotrienols according to the location and number of methyl groups, wherein α -tocopherol has the highest biological activity (Kimberly Kline, Karla. Law son, Wei ping Y u, et al. Vitamin E and Cancer [ J ] Vitamin and Cancer,2007, 76: 435. 461 ]) and is preferentially absorbed and utilized by human body (Zingg J M. E: An organic Vitamin of Vitamin research [ J ]. Molecular Aspects of animal, Medamine, 28(5-6): 400. 422. peanut oil, soybean meal, peanut oil, Vitamin E, peanut oil, peanut.

However, the natural Vitamin E extracted from plants is still insufficient to meet the nutritional and medical requirements, and how to increase the content of Vitamin E (especially α -tocopherol with the highest activity) in plants has become a hotspot of researchers, breeding improvement by genetically engineering genes related to Vitamin E synthesis is a main way and an effective way for increasing the content of Vitamin E in plants at present, the biosynthetic pathway of Vitamin E in plants is that homogentisic acid (HGA) is obtained from the shikimic acid pathway, isopentenyl diphosphate (PrDP) is obtained from the non-mevalonate pathway, and then HGA prenylation is synthesized by a series of enzyme catalysis (Yang W, Cahoon R E, Hunter S C, et al Vitamin E biosynthesis pathway: functional transformation of the monocotylation synthase and transfer enzyme [ J.J.J.865., Plant J.65 (2) and the genetic engineering genes of the transgenic microorganism strain [ 17, 2011. the genetic engineering genes of the transgenic microorganism strain, the genes of the tocopherol, the growth of peanut peroxidase, the transgenic strain No. 2. the transgenic strain No. 7, the transgenic strain No. 7. the transgenic strain can be increased by the genetic engineering genes of the expression of the transgenic microorganism strain No. 23, the transgenic strain No. 2. the transgenic strain No. 7, the transgenic strain No. 2. the transgenic strain can be increased by the genetic engineering genes of the transgenic strain No. 2. the transgenic strain No. 7 strain No. 2. the transgenic strain No. 2, the transgenic strain No. 2. the transgenic strain, the transgenic strain No. 2, the transgenic strain of the transgenic strain can be increased by the genetic engineering genes of the transgenic strain No. 2. the transgenic strain No. 2 strain of the transgenic strain No. 2. the transgenic strain of the transgenic strain No. 2 strain of the transgenic strain A strain of the transgenic strain No. 2 strain A strain of the transgenic strain A strain of the transgenic strain can be obtained by genetic engineering genes of the transgenic strain No. 2 strain of the transgenic strain III strain can be obtained by genetic strain III strain I, the transgenic strain III strain.

In recent years, a sprout food is hot and moist at home and abroad, namely, a food prepared from properly germinated grain seeds serving as raw materials is prepared, and many studies show that germination can improve the utilization rate of grain protein, reduce fat content, improve functional factors and improve the nutritional value of seeds, the royal jelly germinates soybeans in a dark place for 5 days, and finds that α -tocopherol is improved from 2.29mg/100g DW to 7.64mg/100g DW, which is a base value of 3.34 times (the royal jelly, research on special nutritional quality of germinated soybean sprouts, namely [ D ]. southern China university, 2016.) Wufengfengfeng germinates three brown rice varieties in a dark place for 5 days, and finds that vitamin E is improved by 46% -51% compared with that of the brown rice before germination (Wufeng germinates influence on main nutritional ingredients, physiological efficacy and processing characteristics [ D ]. southern China university, 2013 ]), however, the patent of the brown rice with α -tocopherol in peanuts improved by a germination mode is reported in the prior art.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provide a simple, convenient, low-cost and popular method for improving the content of α -tocopherol in peanuts.

The purpose of the invention is realized by the following technical scheme.

A method for increasing α -tocopherol content in peanuts, comprising the following steps:

(1) pretreatment: adding peanut seeds into disinfectant, soaking and disinfecting, repeatedly washing with water until the pH value is neutral, then adding water, and soaking in a dark place;

(2) and (3) germination treatment: and (2) placing the peanut seeds pretreated in the step (1) in a dark condition for germination to obtain peanut sprouts.

Preferably, the peanut seeds in the step (1) are more than one of Yue oil No. 13, Yu Hua No. 15 and Lu Hua No. 11 peanut seeds.

Preferably, the disinfectant in the step (1) is a sodium hypochlorite solution with the concentration of 0.5-1.5 wt%; the soaking and disinfecting time is 5-15 min.

Preferably, the water used for the lightproof soaking in the step (1) is 2-4 times of the mass of the peanut seeds; the temperature for soaking in dark is 20-30 ℃, and the time is 4-12 h.

Preferably, the temperature for germination in the step (2) is 26-32 ℃.

Preferably, the humidity of the germination in the step (2) is 70-100%; and spraying distilled water once every 1-3 hours during the germination period, wherein each spraying lasts for 1-2 minutes. .

Further preferably, the germination time in step (2) is 7 days.

Preferably, the germination time is 7 days.

Preferably, the peanut sprouts in the step (2) are subjected to vacuum freeze drying to enable the moisture content of the peanut sprouts to be 2-3 wt%, then are subjected to high-speed grinding and then are sieved by a 40-60-mesh sieve, and are subjected to vacuum packaging and stored in a drying box at 25 ℃.

Preferably, the α -tocopherol content of the peanut sprout in the step (2) is up to 178.63 mug/g DW-267.52 mug/g DW, and is increased by 4.87-5.98 times compared with the α -tocopherol content in ungerminated peanut seeds.

Preferably, the method for increasing the content of α -tocopherol in peanuts comprises the following specific steps:

(1) pretreatment: selecting high-quality fresh peanut seeds which are full in appearance, consistent in size and free of mildew, cleaning the peanut seeds by using distilled water to remove surface impurities, draining the peanut seeds, soaking the peanut seeds in a 0.5-1.5% sodium hypochlorite solution for 5-15 min for disinfection, repeatedly washing the peanut seeds by using distilled water until the pH value is neutral, adding distilled water with the mass being 2-4 times of that of the peanuts, and soaking the peanut seeds for 4-12 h at 20-30 ℃ in a dark place;

(2) and (3) germination treatment: sprouting in the dark at 26-32 ℃ and 70-100% humidity for 7 days, spraying distilled water once every 1-3 hours, and spraying for 1-2 minutes;

(3) and (3) sample treatment and determination, namely performing vacuum freeze drying on the peanut sprouts to ensure that the moisture content is 2-3%, performing high-speed crushing, sieving by a 40-60-mesh sieve, performing vacuum packaging, placing in a drying box at 25 ℃ for storage, performing high performance liquid chromatography determination under the condition of experimental chromatography, and calculating the α -tocopherol content in the peanut sprouts according to peak areas by using α -tocopherol standard as a reference.

Compared with the prior art, the invention has the following advantages and technical effects:

1. the method is simple, low in cost, popular and suitable for popularization and application;

2. the content of α -tocopherol in the peanuts germinated for 7 days by the method is up to more than 178.63 mug/g DW, and is improved by at least 4.87 times compared with the content of α -tocopherol in ungerminated peanut seeds.

3. The invention is applied to Yue you No. 13, Yu Hua No. 15 and Lu Hua No. 11, and can effectively improve the α -tocopherol content in peanuts.

Drawings

FIG. 1 is a liquid chromatogram of 40 μ g/mL α -tocopherol standard;

FIG. 2 is a liquid chromatogram of Yueji No. 13 peanut of example 1;

FIG. 3 is a liquid chromatogram comparison of germinated peanut No. 13 in Yue oil for 2, 4 and 6 days in example 1.

Detailed Description

The following examples further illustrate the practice of the present invention, but the embodiments are not limited thereto.

Example 1

(1) Pretreatment: selecting high-quality fresh Yue oil No. 13 peanut seeds which are full in appearance, consistent in size and free of mildew, cleaning the seeds with distilled water to remove surface impurities, draining the seeds, soaking the seeds in a sodium hypochlorite solution with the concentration of 1wt% for 10min to sterilize, repeatedly washing the seeds with the distilled water until the pH value is neutral, adding distilled water with the mass 3 times that of peanuts, and soaking the seeds for 6h at 25 ℃ in a dark condition;

(2) and (3) germination treatment: placing the peanuts treated in the step (1) at 27 ℃ and under the condition of 95% humidity in a dark place for germination for 7 days, and spraying distilled water once every 1 hour, wherein each spraying lasts for 1 minute;

(3) sample processing and determination, namely, carrying out vacuum freeze drying on the peanut sprouts obtained in the step (2) to ensure that the moisture content is 2.74wt%, sieving with a 40-mesh sieve, carrying out vacuum packaging, placing in a drying box at 25 ℃ for storage, accurately weighing 2g of peanut sprout powder, adding 10mL of n-hexane, carrying out ultrasonic treatment (40 KHz, 150W) for 30min after vortex oscillation, centrifuging for 15min at 8000rpm, repeating the extraction process for three times, combining the supernatant liquid at 40 ℃ for rotary evaporation, and redissolving with 2mL of chromatographic grade n-hexane, wherein the chromatographic column is a reversed-phase C18 column (150 mm multiplied by 3.9mm, 4 μm, WATERS Nova-Pak C18), the flowing phase is pure methanol, carrying out isocratic elution at the flow rate of 0.9 mL/min, the detector is an ultraviolet detector, the detection wavelength is 294 nm, the column temperature is 40 ℃, the sample amount is 10 μ L, the high performance liquid chromatography is carried out under the experimental conditions, the tocopherol chromatogram is taken as a α -tocopherol standard product, the comparison chromatogram is calculated according to the comparison result that the content of the peanut sprout oil in the peanut sprouts in 7-3513 g of the peanut sprout, the tocopherol content is increased by the comparison chromatogram is found in the comparison chromatogram, the comparison of 7-3513 g of the peanut sprout, the tocopherol standard, the tocopherol peak area is found in the comparison chart of the peanut sprout, the comparison chart 7-3513 g of the peanut sprout, the tocopherol peak area is found by the comparison chart of the comparison chart 7-269.23 g of the tocopherol standard DW 7-3513 g of the peanut sprout, the tocopherol standard.

Example 2

(1) Pretreatment: selecting high-quality fresh Yuhua No. 15 peanut seeds which are full in appearance, consistent in size and free of mildew, cleaning the seeds with distilled water to remove surface impurities, draining the seeds, soaking the seeds in a sodium hypochlorite solution with the concentration of 1.5wt% for 5min for disinfection, repeatedly washing the seeds with distilled water until the pH value is neutral, adding distilled water with the mass of 2 times that of peanuts, and soaking the seeds for 12h at 23 ℃ in a dark place;

(2) and (3) germination treatment: placing the peanuts treated in the step (1) at 32 ℃ and under 100% humidity in a dark place for germination for 7 days, and spraying distilled water once every 2 hours, wherein each spraying lasts for 2 minutes;

(3) and (3) sample treatment and determination, namely, carrying out vacuum freeze drying on the peanut sprouts obtained in the step (2) to ensure that the moisture content is 2.52wt%, carrying out high-speed crushing, sieving with a 40-mesh sieve, carrying out vacuum packaging, and storing in a drying box at 25 ℃, wherein the determination method of the α -tocopherol content in the peanuts is the same as that in example 1, and the detection shows that the α -tocopherol content of ungerminated peanut kernels is 36.68 mu g/g DW, and the α -tocopherol content of the peanut sprouts after 7 days of germination is 178.63 mu g/g DW, and is increased to the background value of 4.87 times.

Example 3

(1) Pretreatment: selecting high-quality fresh Luhua No. 11 peanut seeds which are full in appearance, consistent in size and free of mildew, cleaning the seeds with distilled water to remove surface impurities, draining the seeds, soaking the seeds in a sodium hypochlorite solution with the concentration of 0.5 wt% for 15min for disinfection, repeatedly washing the seeds with distilled water until the pH value is neutral, adding distilled water with the mass of 4 times that of peanuts, and soaking the seeds for 4h at 27 ℃ in a dark condition;

(2) and (3) germination treatment: placing the peanuts treated in the step (1) at 29 ℃ and under 90% humidity in a dark place for germination for 7 days, and spraying distilled water once every 3 hours, wherein each spraying lasts for 2 minutes;

(3) and (3) sample treatment and determination, namely, carrying out vacuum freeze drying on the peanut sprouts obtained in the step (2) to ensure that the moisture content is 2.48wt%, carrying out high-speed grinding, then sieving with a 60-mesh sieve, carrying out vacuum packaging, and storing in a drying box at 25 ℃, wherein the determination method of α -tocopherol content in the peanuts is the same as that in example 1, and the detection shows that the α -tocopherol content of ungerminated peanut kernels is 50.28 mu g/g DW, and the α -tocopherol content of the peanut sprouts after 7 days of germination is 272.52 mu g/g DW, and is increased to the background value of 5.42 times.

Claims

1. a method for improving α-tocopherol content in peanut, is characterized in that, comprises the following steps:

(1) Pre-treatment: Add peanut seeds into disinfectant, soak them for disinfection, rinse with water repeatedly until the pH value is neutral, then add water and soak in the dark;

(2) Germination treatment: the peanut seeds after the pretreatment in step (1) are placed under light-proof conditions to germinate to obtain peanut buds;

In the step (1), the water used for soaking in the dark is 2 to 4 times the quality of the peanut seeds; the temperature for soaking in the dark is 20 to 30°C, and the time is 4 to 12 hours;

The temperature of the germination in step (2) is 26-32°C;

The humidity of the germination in step (2) is 70-100%;

During the germination of step (2), distilled water is sprayed once every 1 to 3 hours, and each spray lasts for 1 to 2 minutes;

The germination time of step (2) is 7 days.

2. a kind of method that improves α-tocopherol content in peanut according to claim 1, is characterized in that, the peanut seeds described in step (1) are Yueyou No. 13, Yuhua No. 15 and Luhua No. 11 peanuts more than one of the seeds.

3. The method for improving the content of α-tocopherol in peanuts according to claim 1, wherein the disinfectant solution in step (1) is a sodium hypochlorite solution with a concentration of 0.5 to 1.5wt%; the immersion disinfection The time is 5~15min.

4. a kind of method that improves α-tocopherol content in peanut according to claim 1, it is characterised in that the peanut bud described in step (2) is vacuum freeze-dried, so that the peanut bud moisture content is 2～3 wt%, then pulverized and passed through a 40-60 mesh sieve, and finally vacuum-packed and stored in a drying oven at 25°C.

5. The method for increasing the content of α-tocopherol in peanuts according to claim 1, wherein the content of α-tocopherol in the peanut sprouts of step (2) is as high as 178.63 μg/g DW ~ 267.52 μg/ g DW, compared with the content of α-tocopherol in ungerminated peanut seeds, increased by 4.87-5.98 times.