LU101609B1

LU101609B1 - Method for enriching lutein and zeaxanthin by utilizing germinated corns

Info

Publication number: LU101609B1
Application number: LU101609A
Authority: LU
Inventors: Hao Luo; Zhongyuan Zhang; Jiangfeng Song; Lei Feng; Dajing Li; Chunquan Liu; Weiwei He
Original assignee: Jiangsu Acad Agricultural Sci
Priority date: 2019-08-19
Filing date: 2019-08-19
Publication date: 2021-07-01
Also published as: LU101609A1

Abstract

The present invention provides a method for enriching lutein and zeaxanthin by utilizing germinated corns. According to the method provided by the invention,corn seeds with a germinating capability are taken as raw materials, disinfected with a sodium hypochlorite solution, sequentially soaked with a methyl jasmonate solution and a sodium chloride solution, darkly cultivated at 25°C to 28°C for 48 hours to 96 hours, and then sprayed with the methyl jasmonate solution every 4 hours to 6 hours in a germinating process. Upon determination, the method provided by the invention can improve a lutein content in the germinated corn by more than 50% and a zeaxanthin content in the germinated corn by more than 40%. The invention also provides a germinated corn prepared based on the method and an application of the germinated corn in preparing food, Pharmaceuticals, health care products or cosmetics containing lutein.

Description

METHOD FOR ENRICHING LUTEIN AND ZEAXANTHIN BY 191609 : UTILIZING GERMINATED CORNS

TECHNICAL FIELD The present invention relates to the field of development and utilization of biological resources, and more particularly, to a method for enriching lutein and zeaxanthin by utilizing germinated corns. | BACKGROUND Lutein and zeaxanthin are oxygen-containing carotenoids, which contain a plurality of unsaturated double bonds and ionone rings, widely exist in nature and are the main | components of plant pigments such as vegetables, fruits, flowers, etc. Meanwhile, the | lutein and the zeaxanthin are also the major pigments in macula retinae regions of human eyes. Studies have found that the lutein has certain preventive and improvement | 15 effects on eye health care, diabetes mellitus, cardiovascular diseases, cancers and a ! plurality of other chronic diseases. The zeaxanthin has a stronger antioxidant capacity, | which can protect tissue cells of an organism by antioxidant behaviors such as quenching singlet oxygen and scavenging free radicals, so as to reduce the outbreak risk of some diseases. Therefore, the development of food rich in the lutein and the zeaxanthin has become a hot spot and has been allowed to be added into food, beverages, | cosmetics and health products as a food supplement. Studies on a production process of lutein in the prior art are mainly reflected in the | studies on how to separate and purify raw materials. For example, the patent (Publication Number of CN103058904B and Publication Date of March 30, 2016) ; 25 discloses a novel method for enriching and purifying lutein in marigold, in which supercritical CO2 extraction and membrane separation technologies are utilized to | enrich and purify the lutein in the marigolds. However, there are few studies on enriching the lutein by biological methods at present. | After sprouting, chemical compositions of plant seeds are all changed, contents of active substances are increased, and unique flavor and taste are formed, thus improving lu101609 | a nutritional quality of the plants. The patent (Publication Number of CN102754780B | and Publication Date of March 5, 2014) discloses "Gama-aminobutyric Acid-enriched | Freeze-drying Bean Sprout and Production Technique Thereof". The patent (Publication Number of CN107211852A and Publication Date of September 29, 2017) discloses "Production Method for Enriching Gama-aminobutyric Acid in Barley". The patent (Publication Number of CN106922509A and Publication Date of July 7, 2017) ; discloses "Production Method for Enriching Gamma-aminobutyric Acid in Bean Sprouts and Increasing Yield". The patent (Publication Number of CN105901519A and ; 10 Publication Date of August 31, 2016) discloses "Method of Increasing Content of i Gamma-aminobutyric Acid in Coarse Millet". The patent (Publication Number of ; CN104498548A and Publication Date of April 8, 2015) discloses "Production Method ; for Enriching Gama-aminobutyric Acid by Utilizing Germinated Brown Rice". The | above patents only use seed seeds to germinate and enrich the gamma-aminobutyric ; 15 acid, but none of these patents published the simultaneous enrichment of the lutein and the zeaxanthin by corn germination. As an important food crop in China, the corns have a wide range of raw materials and a great development value in the fields of nutrition ; and health care.

SUMMARY / An object of the present invention is to provide a method for enriching lutein and ; zeaxanthin by utilizing germinated corns.

The present invention provides a method for enriching lutein and zeaxanthin by | utilizing germinated corns, wherein corn seeds with a germinating capability are taken ; 25 as raw materials, disinfected with a sodium hypochlorite solution, sequentially soaked ; with a methyl jasmonate solution and a sodium chloride solution, darkly cultivated at a | constant temperature, and then the germinated corn seeds are sprayed with the methyl | jasmonate solution every 4 hours to 6 hours during the period.

| Preferably, a concentration of the sodium hypochlorite solution used for disinfecting . 30 the com seeds is 0.5%, and the disinfection lasts for 20 minutes to 30 minutes. a EET

| ; [ Preferably, when soaking the corn seeds, the methyl jasmonate solution used is 0.1 lu101609 | pmol/L to 10.0 pmol/L, and the corn seeds are soaked with the methyl jasmonate É solution for 15 hours to 20 hours.

Preferably, when soaking the corn seeds, the sodium chloride solution used is 300 mmol/L, and the corn seeds are soaked with the sodium chloride solution for 6 hours to hours.

Preferably, a temperature of the dark cultivation is 25°C.

| Preferably, the dark cultivation lasts for 72 hours.

! Preferably, the methyl jasmonate solution spayed during the germinating process is 0.1 j 10 pmol/L to 10.0 umol/L, and the methyl jasmonate solution is evenly spayed once every ; 4 hours to 6 hours.

, The present invention also provides a germinated corn prepared by the method / mentioned above, wherein based on a dry weight of the germinated corn, a lutein ; content in the germinated corn is no less than 12 ug/g, and a zeaxanthin content in the germinated corn is no less than 9 pg/g.

The present invention also provides an application of the germinated corn in preparing | food, pharmaceuticals, health care products or cosmetics containing lutein.

, Beneficial effects: (1) According to the present invention, the corn seeds are used as the raw materials, and | 20 exogenous methyl jasmonate with an appropriate concentration is applied before and ; during soaking the corn seeds. The methyl jasmonate is an important plant signal | substance, and participates in various development processes such as seed germination, | root system growth, generation, fruit ripening, senescence and the like in plants, which ; can increase the resilience of the corn seeds to adversity, and improve a growth . 25 condition of the corn seeds under adversity stress; and promote synthesis of various : amino acids and proteins and enhance accumulation of antioxidant substances. The | addition of the sodium chloride will produce adversity stress on the plants, which is not conducive to the growth of the plants, but can induce enrichment of secondary ; metabolites such as carotenoids and phenols, and increase antioxidant capacities of ; 30 those secondary metabolites. Spraying the methyl jasmonate during the germinating

‘ | can maintain a defense mechanism of the corn seeds and offset adverse effects on ‘101608 growth caused by sodium chloride stress.

| (2) According to the method provided by the present invention, the methyl jasmonate | and the sodium chloride are used to soak the corn seeds in sequence, which is | 5 convenient to process, low in cost, safe and non-toxic, and easy for large-scale | production and use.

(3) In the present invention, the corn seeds are subjected to the joint treatment of being | soaked with the methyl jasmonate and the sodium chloride and sprayed with the methyl 2 jasmonate, which has corresponding promoting effects on physiological characteristics such as sprout length, root length and the like of the germinated corns, can improve a ; germination rate, a sprout length and an antioxidant capacity of the corn seeds, and | improve the nutritional quality and health care functions of the product.

; Other advantages, objects and features of the invention will be partially reflected by the | following description, and will be partially understood by those skilled in the art ; 15 through research and practice of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS | Fig. 1 is a change of lutein contents in a germinated corn seed jointly treated with methyl jasmonate of different concentrations and 300 mmol/L sodium chloride in a method of the present invention; and | Fig. 2 is a change of zeaxanthin contents in the germinated corn seed jointly treated : with methyl jasmonate of different concentrations and 300 mmol/L sodium chloride in : the method of the present invention.

: 25 DETAILED DESCRIPTION The invention will be further described in detail hereinafter with reference to the | accompanying drawings, so that those skilled in the art can implement the invention . with reference to the description.

; It should be understood that terms such as "having," "including," and "comprising" as | 30 used herein do not exclude the presence or addition of one or more other elements or

| combinations thereof. 101609 | The present invention provides a method for enriching lutein by utilizing germinated | coms, wherein corn seeds with a germinating capability are taken as raw materials, ) disinfected with a sodium hypochlorite solution, sequentially soaked with a methyl | 5 jasmonate solution and a sodium chloride solution, darkly cultivated at a constant temperature, and then the germinated corn seeds are sprayed with the methyl jasmonate : solution during the germinating period.

Before soaking, the corn seeds with the germinating capability are preferably screened according to the present invention, and the corn seeds are preferably washed with clear | 10 water to remove impurities during the screening. Then the corn seeds are put in the clear | water, and floating corn seeds are removed after 5 minutes to 10 minutes. In the present ) invention, the purpose of the screening is mainly to remove the impurities in the corn | seeds and the corn seeds which are not plumpy.

| According to the present invention, the screened corn seeds are preferably disinfected, | 15 and the sodium hypochlorite solution is preferably used to soak the corn seeds during | the disinfection. A concentration of the sodium hypochlorite solution is preferably 0.1% ; to 1%, and more preferably 0.5%. The disinfection preferably lasts for 10 minutes to 30 minutes, and more preferably lasts for 20 minutes. In the present invention, deionized ‘ water is preferably used to wash the sterilized corn seeds to clean the residual sodium ; 20 hypochlorite solution on surfaces of the corn seeds until a pH value on the surfaces of ; the com seeds is 7.

In the present invention, the corn seeds with the germinating capability are soaked with the methyl jasmonate solution. In the present invention, a concentration of the methyl | jasmonate solution is preferably 0.1 um ol/l to 10.0. pm ol/l, and more preferably 1 um ol/l. The corn seeds are soaked for preferably 15 hours to 20 hours, and more preferably | 18 hours. The corn seeds are soaked preferably under a temperature of 20°C to 30°C, and more preferably 25°C. An application of exogenous methyl jasmonate with appropriate concentration during the seed soaking can induce a defense reaction of plants, and improve a growth status of the plants and resistance of the plants to adversity. | 30 After soaking the corn seeds with the methyl jasmonate solution, distilled water is | preferably used to clean the residual solution on the surfaces of the corn seeds according

: 6 to the present invention, and then the corn seeds are soaked with sodium chloride, (101608 ; wherein the sodium chloride solution is 300 mmol/L, and the corn seeds are soaked l with the sodium chloride solution for 6 hours to 10 hours, and more preferably 8 hours. ; After soaking the corn seeds with the sodium chloride solution, distilled water is ; 5 preferably used to clean the residual solution on the surfaces of the corn seeds, and the | cleaned corn seeds are placed in a constant-temperature constant-humidity incubator : for dark cultivation. In the present invention, a temperature of the dark culture 1s 25°C | to 28°C, and preferably 25°C. A relative humidity of the dark cultivation is preferably 70% to 98% and more preferably 80% to 95%. The dark cultivation preferably lasts for ; 10 24 hours to 96 hours, and more preferably 72 hours. The purpose of culturing the corn | seeds in the dark for 24 hours to 96 hours is to enable the corn seeds to germinate ‘ naturally, improve a growing ability of the corn seeds, maintain a certain germinating ) rate, and fully enrich nutrients such as lutein, zeaxanthin and the like.

; In the dark cultivation process, the corn seeds are preferably placed in a culture dish ; 15 which is padded with two layers of filter paper wetted by distilled water, and sprayed with the methyl jasmonate solution every 4 hours to 6 hours, wherein a concentration of the methyl jasmonate solution is preferably 0.1 pmol/l to 10.0 umol/l, and more ; preferably 1 umol/L, so as to keep the germinated corns moist, maintain moisture | required for normal growth, and promote large accumulation of lutein and zeaxanthin. | 20 In the embodiment of the present invention, lutein and zeaxanthin contents in germinated corn freeze-dried powder are also determined. Based on a dry weight of the germinated corn, the lutein content in the germinated corn is rich, which is no less than : 12 ug/g, and the zeaxanthin content in the germinated corn is no less than 9 ug/g.

The present invention also provides an application of the germinated corns in preparing | 25 food, pharmaceuticals, health care products or cosmetics containing lutein, which uses ; the germinated corns mentioned above to match with other active ingredients, or, use ; the germinated corns mentioned above as the unique active ingredient to match with | carriers acceptable in the field to prepare foods, pharmaceuticals, health care products ; or cosmetics. A dosage of the germinated corn is 1% to 100%.

The technical solutions in the embodiments of the invention will be clearly and | completely described below with reference to the embodiments of the invention. Based

: on the embodiments of the invention, other embodiments obtained by those having lu101609 . ordinary skills in the art without going through any creative work shall all fall within ; the scope of protection of the invention.

j Embodiment 1 | 5 (1) Screening and disinfecting corn seeds: after removing impurities in the corn seeds ‘ ; and the corn seed which were not plumpy, the corn seeds were washed with clear water, ; stood for 5 minutes to 10 minutes, floating corn seeds were removed, and then soaked . in 0.5% sodium hypochlorite solution for disinfection for 20 minutes.

| (2) The corn seeds in the step (1) were washed with deionized water to clean the residual | 10 sodium hypochlorite solution on surfaces of the corn seeds.

(3) Soaking the corn seeds with methyl jasmonate: the corn seeds were soaked with 1 | pmol/L methyl jasmonate solution at normal temperature for 16 hours. After soaking ; the corn seeds, the residual solution on the surfaces ofthe corn seeds were cleaned with ) deionized water.

(4) Soaking the corn seeds with sodium chloride: the corn seeds were soaked with 300 | mmol/L sodium chloride solution at normal temperature for 8 hours.

; (5) Germinating the corn seeds: every 30 g (20 to 25 seeds) of the corn seeds in the step | (4) were put into a culture dish (®15 cm) which was padded with two layers of wetted | filter paper, and placed in a constant temperature incubator at 25°C for dark germination | 20 for 72 hours.

} (6) Spraying methyl jasmonate: in the germinating process of the germinate corns in the step (4), 30 mL to 40 mL of 1 umol/L methyl jasmonate solution were sprayed every | 4 hours to keep the germinated corns wet to germinate.

: (7) Drying and pulverizing the corn seeds: the germinated corn seeds and sprouts | 25 thereof were washed and drained off, put into a refrigerator at 80°C below zero to pre- freeze for 24 hours, dried by a vacuum freeze dryer, and then crushed and sieved | through a 60-mesh sieve, wherein a lutein content and a zeaxanthin content were measured to be 13.96 ug/g and 9.66 ug/g respectively.

| Embodiment 2 | 30 (1) Screening and disinfecting corn seeds: after removing impurities in the corn seeds

: and the corn seed which were not plumpy, the corn seeds were washed with clear water, u101609 ; stood for 5 minutes to 10 minutes, floating com seeds were removed, and then soaked : in 0.5% sodium hypochlorite solution for disinfection for 25 minutes.

; (2) The corn seeds in the step (1) were washed with deionized water to clean the residual . 5 sodium hypochlorite solution on surfaces of the corn seeds.

| (3) Soaking the corn seeds with methyl jasmonate: the corn seeds were soaked with 5 | pmol/L methyl jasmonate solution at normal temperature for 16 hours. After soaking | the corn seeds, the residual methyl jasmonate solution on the surfaces of the corn seeds ) were cleaned with deionized water.

| 10 (4) Soaking the corn seeds with sodium chloride: the corn seeds were soaked with 300 mmol/L sodium chloride solution at normal temperature for 8 hours.

(5) Germinating the corn seeds: every 30 g (20 to 25 seeds) of the corn seeds in the step | (4) were put into a culture dish (®15 cm) which was padded with two layers of wetted filter paper, and placed in a constant temperature incubator at 25°C for dark germination ; 15 for 12 hours.

, (6) Spraying methyl jasmonate: in the germinating process of the germinate corns in the step (4), 30 mL to 40 mL of 5 pmol/L methyl jasmonate solution were sprayed every | 4 hours to keep the germinated corns wet to germinate.

/ (7) Drying and pulverizing the corn seeds: the germinated corn seeds and sprouts | 20 thereof were washed and drained off, put into a refrigerator at 80°C below zero to pre- / freeze for 24 hours, dried by a vacuum freeze dryer, and then crushed and sieved / through a 60-mesh sieve, wherein a lutein content and a zeaxanthin content were | measured to be 13.38 ug/g and 9.55 ug/g respectively.

| Embodiment 3 (1) Screening and disinfecting corn seeds: after removing impurities in the corn seeds and the corn seed which were not plumpy, the corn seeds were washed with clear water, | stood for 5 minutes to 10 minutes, floating corn seeds were removed, and then soaked | in 0.5% sodium hypochlorite solution for disinfection for 25 minutes.

; (2) The corn seeds in the step (1) were washed with deionized water to clean the residual | 30 sodium hypochlorite solution on surfaces of the corn seeds. DEE :

| (3) Soaking the corn seeds with methyl jasmonate: the corn seeds were soaked with 10 u101609 i pmol/L methyl jasmonate solution at normal temperature for 16 hours. After soaking ] the corn seeds, the residual methyl jasmonate solution on the surfaces of the corn seeds | were cleaned with deionized water.

; 5 (4) Soaking the corn seeds with sodium chloride: the corn seeds were soaked with 300 | mmol/L sodium chloride solution at normal temperature for 8 hours.

(5) Germinating the corn seeds: every 30 g (20 to 25 seeds) of the corn seeds in the step ; (4) were put into a culture dish (®15 cm) which was padded with two layers of wetted | filter paper, and placed in a constant temperature incubator at 25°C for dark germination for 12 hours.

(6) Spraying methyl jasmonate: in the germinating process of the germinate corns in | the step (4), 30 mL to 40 mL of 5 umol/L methyl jasmonate solution were sprayed every : 4 hours to keep the germinated corns wet to germinate.

| (7) Drying and pulverizing the corn seeds: the germinated corn seeds and sprouts | 15 thereof were washed and drained off, put into a refrigerator at 80°C below zero to pre- | freeze for 24 hours, dried by a vacuum freeze dryer, and then crushed and sieved through a 60-mesh sieve, wherein a lutein content and a zeaxanthin content were | measured to be 12.85 pg/g and 9.08 ug/g respectively.

| Comparison Example 1 | 20 Compared with Embodiment 1, the steps (3), (4) and (6) were omitted, and the corn : seeds soaked with the sodium chloride were directly placed in a constant temperature | incubator at 25°C for dark germination for 72 hours.

! It can be seen from Fig. 1 and Fig. 2: after drying and pulverizing the germinated corns | treated by the solution described in the Comparison Example 1, a lutein content was | 25 measured to be 8.29 ug/g and a zeaxanthin content was measured to be 6.09 ug/g/g.

The lutein contents of Embodiments 1 to 3 were increased by 68.40%, 61.40% and ; 55.00% respectively, and the zeaxanthin contents were increased by 58.62%, 56.81% | and 49.10% respectively in comparison with the Comparison Example 1.

| The embodiments of the invention have been described in detail above, but these are | 30 merely examples for ease of understanding and should not be considered as limiting the

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| 10

; scope of the invention.

Similarly, any person skilled in the art can make various possible 1101609

; equivalent changes or substitutions according to the description of the technical ı solutions of the invention and preferred embodiments thereof, but all these changes or f substitutions shall fall within the scope of protection of the claims of the invention. re RE OOo ES ;

Claims

: lu101609 / Claims / 1. A method for enriching lutein and zeaxanthin by utilizing germinated corns, : wherein corn seeds with a germinating capability are taken as raw materials, disinfected | with a sodium hypochlorite solution, sequentially soaked with a methyl jasmonate ; solution and a sodium chloride solution, darkly cultivated at 25°C to 28°C for 48 hours | to 96 hours, and then sprayed with the methyl jasmonate solution every 4 hours to 6 / hours in a germinating process,
2. The method according to claim 1, wherein a concentration of the sodium hypochlorite solution used for disinfecting the corn seeds is 0.5%, and the disinfection lasts for 20 minutes to 30 minutes.
3. The method according to claim 1, wherein when soaking the corn seeds, the methyl] jasmonate solution used is 0.1 pmol/L to 10.0 pmol/L, and the corn seeds are soaked with the methyl jasmonate solution for 15 hours to 20 hours.
4. The method according to claim 1, wherein when soaking the corn seeds, the sodium chloride solution used is 300 mmol/L, and the corn seeds are soaked with the sodium chloride solution for 6 hours to 10 hours. |
5. The method according to claim 1, wherein a temperature of the dark cultivation is 25°C.
6. The method according to claim 5, wherein the dark cultivation lasts for 72 hours.
7. The method according to claim 1, wherein the methyl jasmonate solution spayed | | during the germinating process is 0.1 pmol/L to 10.0 umol/L, and the methyl jasmonate solution is evenly spayed once every 4 hours to 6 hours. | | |

: lu101609
8. A germinated corn prepared by the method according to claims 1 to 7, wherein : based on a dry weight of the germinated corn, a lutein content in the germinated corn is no less than 12 ug/g, a zeaxanthin content in the germinated corn is no less than 9 ug/g, and a sprout length is no less than 2.5 cm.
9. An application of the germinated corn according to claim 8 in preparing food, | pharmaceuticals, health care products or cosmetics containing lutein. ee pee picid eee