KR102457241B1 - Method for production of sulforaphane from Brassicaceae plant with phenylisothiocynate and method for preservation of sulforaphane - Google Patents
Method for production of sulforaphane from Brassicaceae plant with phenylisothiocynate and method for preservation of sulforaphane Download PDFInfo
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- SUVMJBTUFCVSAD-UHFFFAOYSA-N sulforaphane Chemical compound CS(=O)CCCCN=C=S SUVMJBTUFCVSAD-UHFFFAOYSA-N 0.000 title claims abstract description 162
- SUVMJBTUFCVSAD-JTQLQIEISA-N 4-Methylsulfinylbutyl isothiocyanate Natural products C[S@](=O)CCCCN=C=S SUVMJBTUFCVSAD-JTQLQIEISA-N 0.000 title claims abstract description 81
- 229960005559 sulforaphane Drugs 0.000 title claims abstract description 81
- 235000015487 sulforaphane Nutrition 0.000 title claims abstract description 81
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 17
- QKFJKGMPGYROCL-UHFFFAOYSA-N phenyl isothiocyanate Chemical compound S=C=NC1=CC=CC=C1 QKFJKGMPGYROCL-UHFFFAOYSA-N 0.000 title claims description 29
- 238000000034 method Methods 0.000 title claims description 20
- 238000004321 preservation Methods 0.000 title claims 2
- 241000219193 Brassicaceae Species 0.000 title 1
- 108010058651 thioglucosidase Proteins 0.000 claims abstract description 24
- 150000001413 amino acids Chemical class 0.000 claims abstract description 17
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 17
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 17
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 17
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 14
- 239000000203 mixture Substances 0.000 claims abstract description 6
- 241000196324 Embryophyta Species 0.000 claims description 33
- 235000011299 Brassica oleracea var botrytis Nutrition 0.000 claims description 23
- 235000017647 Brassica oleracea var italica Nutrition 0.000 claims description 23
- 240000003259 Brassica oleracea var. botrytis Species 0.000 claims description 23
- 239000006228 supernatant Substances 0.000 claims description 12
- RUQCCAGSFPUGSZ-OBWQKADXSA-N Glucoraphanin Natural products C[S@](=O)CCCCC(=NS(=O)(=O)O)S[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O RUQCCAGSFPUGSZ-OBWQKADXSA-N 0.000 claims description 10
- GMMLNKINDDUDCF-JRWRFYLSSA-N [(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] (1e)-5-[(r)-methylsulfinyl]-n-sulfooxypentanimidothioate Chemical compound C[S@@](=O)CCCC\C(=N/OS(O)(=O)=O)S[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O GMMLNKINDDUDCF-JRWRFYLSSA-N 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 7
- 229940117953 phenylisothiocyanate Drugs 0.000 claims description 6
- 238000005119 centrifugation Methods 0.000 claims description 5
- 235000021537 Beetroot Nutrition 0.000 claims description 4
- 240000007124 Brassica oleracea Species 0.000 claims description 4
- 235000003899 Brassica oleracea var acephala Nutrition 0.000 claims description 4
- 235000011301 Brassica oleracea var capitata Nutrition 0.000 claims description 4
- 235000001169 Brassica oleracea var oleracea Nutrition 0.000 claims description 4
- 244000304217 Brassica oleracea var. gongylodes Species 0.000 claims description 4
- 150000001875 compounds Chemical class 0.000 abstract description 24
- 125000000524 functional group Chemical group 0.000 abstract description 16
- 239000012038 nucleophile Substances 0.000 description 8
- 239000000523 sample Substances 0.000 description 6
- IQPQWNKOIGAROB-UHFFFAOYSA-N isocyanate group Chemical group [N-]=C=O IQPQWNKOIGAROB-UHFFFAOYSA-N 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 2
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 238000010298 pulverizing process Methods 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 238000001291 vacuum drying Methods 0.000 description 2
- -1 PITC compound Chemical class 0.000 description 1
- 206010033546 Pallor Diseases 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 230000007247 enzymatic mechanism Effects 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 150000002540 isothiocyanates Chemical class 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
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- C12P13/00—Preparation of nitrogen-containing organic compounds
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- C07C331/00—Derivatives of thiocyanic acid or of isothiocyanic acid
- C07C331/16—Isothiocyanates
- C07C331/18—Isothiocyanates having isothiocyanate groups bound to acyclic carbon atoms
- C07C331/22—Isothiocyanates having isothiocyanate groups bound to acyclic carbon atoms of an unsaturated carbon skeleton
- C07C331/24—Isothiocyanates having isothiocyanate groups bound to acyclic carbon atoms of an unsaturated carbon skeleton the carbon skeleton containing six-membered aromatic rings
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- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01147—Thioglucosidase (3.2.1.147), i.e. myrosinase
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Abstract
본 발명은 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물을 처리함으로써 설포라판이 화합물화되는 것을 방지하여 설포라판을 생산할 수 있는 방법에 관한 것으로, 본 발명에서는 십자화과 식물의 분쇄액에 과량의 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물을 처리하여 십자화과 식물 내의 단백질, 펩타이드 및 아미노산 중 선택되는 어느 하나 이상을 침전시켜 설포라판이 화합물화되는 것을 원천적으로 막았고, 마이로시네이즈를 처리함으로써, 설포라판을 고순도로 수득할 수 있었다. 또한, 본 발명은 단백질, 펩타이드 및 아미노산 중 선택되는 어느 하나 이상과 설포라판을 함유하는 혼합물에 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물을 처리함으로써, 설포라판을 장시간 안정적으로 보존할 수 있었다. The present invention relates to a method for producing sulforaphane by preventing sulforaphane from being compounded by treating a compound containing an isothiocynate functional group. Treating a compound containing an isothiocynate functional group to precipitate any one or more selected from proteins, peptides and amino acids in cruciferous plants to fundamentally prevent sulforaphane from being compounded, and by treating myrosinase, sulforaphane with high purity could be obtained with In addition, the present invention was able to stably preserve sulforaphane for a long time by treating a compound containing an isothiocynate functional group in a mixture containing any one or more selected from proteins, peptides and amino acids and sulforaphane.
Description
본 발명은 십자화과 식물로부터 설포라판을 생산하는 방법 및 보존하는 방법에 관한 것으로, 더욱 상세하게는 페닐이소티오시아네이트를 처리함으로써 설포라판이 화합물화되는 것을 방지하여 십자화과 식물로부터 설포라판을 고수율로 생산할 수 있는 방법 및 설포라판을 고순도로 보존할 수 있는 방법에 관한 것이다. The present invention relates to a method for producing and preserving sulforaphane from a cruciferous plant, and more particularly, to a method for producing sulforaphane from a cruciferous plant in high yield by treating phenyl isothiocyanate to prevent sulforaphane from being compounded. It relates to a method and a method capable of preserving sulforaphane in high purity.
브로콜리, 콜라비, 레드비트, 양배추와 같은 십자화과 식물에는 당과 합성된 배당체 형태의 글루코시놀레이트(glucosinolate) 계의 글루코라파닌(glucoraphanin)이 존재한다. 글루코라파닌은 마이로시네이즈(myrosinase)라는 효소에 의해 에 의해 당과 비당으로 가수분해되어, 항산화, 항암 등의 기능성이 있는 것으로 보고된 이소티오시아네이트(isothiocyanate) 계의 설포라판(sulforaphane)으로 생성된다. In cruciferous plants such as broccoli, kohlrabi, red beet, and cabbage, there is glucoraphanin in the form of a glycoside synthesized with sugar. Glucorapanin is hydrolyzed to sugar and non-saccharide by an enzyme called myrosinase, and is an isothiocyanate-based sulforaphane reported to have antioxidant and anticancer functions. is created
그런데, 마이로시네이즈(myrosinase) 효소에 의해 설포라판이 생성되면, 십자화과 식물 내의 뉴클레오파일(nucleophile, 대부분의 단백질, 펩타이드 또는 아미노산들)들이 설포라판과 화합물(sulforaphane conjugate)을 이루어 설포라판의 수득에 부정적인 영향을 주는 문제가 있다. However, when sulforaphane is produced by myrosinase enzyme, nucleophiles in cruciferous plants (nucleophile, most proteins, peptides or amino acids) form a compound (sulforaphane conjugate) with sulforaphane, which is negative for obtaining sulforaphane There are issues that affect it.
이에 본 발명에서는 이와 같은 문제를 해결하여 설포라판을 고수율로 수득할 수 있는 방법을 개발하고자 한다. Therefore, in the present invention, it is intended to develop a method capable of obtaining sulforaphane in high yield by solving such a problem.
십자화과 식물에는 설포라판(sulforaphane)의 기질(substrate)인 글르코라파닌(glucoraphanin)이 다량 포함되어 있는데, 십자화과 식물 자체 내의 마이로시네이즈(myrosinase) 효소에 의해 설포라판이 생성되면, 십자화과 식물 내의 뉴클레오파일(대부분의 단백질, 펩타이드 또는 아미노산들)이 설포라판과 화합물(sulforaphane conjugate)을 이루어 설포라판의 수득에 부정적인 영향을 준다. 본 발명에서는 이와 같은 문제점을 해결하여 십자화과 식물로부터 설포라판을 고수율로 수득할 수 있는 방법을 개발하여 제공하고자 한다. Cruciferous plants contain a large amount of glucoraphanin, a substrate of sulforaphane, and when sulforaphane is produced by myrosinase enzyme in the cruciferous plant itself, nucleos in the cruciferous plant Pile (most proteins, peptides or amino acids) forms a compound (sulforaphane conjugate) with sulforaphane, which negatively affects the yield of sulforaphane. In the present invention, by solving such a problem, it is intended to develop and provide a method for obtaining sulforaphane in high yield from cruciferous plants.
또한, 본 발명은 설포라판을 고순도로 보존할 수 있는 방법을 개발하여 제공하고자 한다. In addition, the present invention is to develop and provide a method for preserving sulforaphane in high purity.
본 발명은 십자화과 식물의 분쇄액에 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물를 처리하는 단계 (a); 상기 단계 (a) 후, 마이로시네이즈(myrosinase)를 처리하는 단계 (b);를 포함하는 것을 특징으로 하는 설포라판(sulforaphane)의 생산방법을 제공한다. The present invention comprises the steps of treating a compound containing an isothiocynate functional group to a pulverized solution of a cruciferous plant (a); After the step (a), the step (b) of treating myrosinase (myrosinase); provides a method for producing sulforaphane (sulforaphane) comprising the.
본 발명의 설포라판(sulforaphane) 생산방법에 있어서, 상기 십자화과 식물은, 일 예로 브로콜리, 콜라비, 레드비트, 양배추 중 선택되는 어느 하나일수 있다. In the method for producing sulforaphane of the present invention, the cruciferous plant may be, for example, any one selected from broccoli, kohlrabi, red beet, and cabbage.
본 발명의 설포라판(sulforaphane) 생산방법에 있어서, 상기 십자화과 식물의 분쇄액에는, 바람직하게 글루코라파닌(glucoraphanin)이 함유되어 있을 수 있다.In the method for producing sulforaphane of the present invention, the crushed liquid of the cruciferous plant may preferably contain glucoraphanin.
본 발명의 설포라판(sulforaphane) 생산방법에 있어서, 상기 단계 (a)는, 바람직하게 아이소사이오시아네이트(isothiocynate)의 처리에 의해 십자화과 식물 분쇄액 내의 단백질, 펩타이드 및 아미노산 중 선택되는 어느 하나 이상이 침전되는 것일 수 있다. In the method for producing sulforaphane of the present invention, in step (a), at least one selected from proteins, peptides and amino acids in the crushed cruciferous plant is precipitated, preferably by treatment with isothiocynate. it may be
본 발명의 설포라판(sulforaphane) 생산방법에 있어서, 상기 설포라판의 생산방법은, 바람직하게 상기 단계 (a) 후, 원심분리하여 상등액을 회수한 후, 회수한 상등액에 단계 (b)의 마이로시네이즈(myrosinase)를 처리하는 것이 좋다. In the method for producing sulforaphane of the present invention, the method for producing sulforaphane is preferably, after step (a), centrifuging to recover the supernatant, and then adding the myrosinase of step (b) to the recovered supernatant. (myrosinase) treatment is recommended.
한편, 본 발명은 단백질, 펩타이드 및 아미노산 중 선택되는 어느 하나 이상과 설포라판을 함유하는 혼합물에 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물을 처리하는 것을 특징으로 하는 설포라판의 보존방법을 제공한다. Meanwhile, the present invention provides a method for preserving sulforaphane, comprising treating a compound containing an isothiocynate functional group to a mixture containing at least one selected from protein, peptide and amino acid and sulforaphane.
본 발명에서는 십자화과 식물의 분쇄액에 과량의 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물을 처리하여 십자화과 식물 내의 단백질, 펩타이드 및 아미노산 중 선택되는 어느 하나 이상을 침전시켜 원천적으로 설포라판이 화합물화되는 것을 막았고, 마이로시네이즈를 처리함으로써 설포라판을 고수율로 수득할 수 있었다. In the present invention, sulforaphane is originally compounded by precipitating any one or more selected from proteins, peptides and amino acids in cruciferous plants by treating a compound containing an excess of isothiocynate functional groups in the crushed liquid of a cruciferous plant. was prevented, and sulforaphane was obtained in high yield by treatment with myrosinase.
또한, 본 발명은 단백질, 펩타이드 및 아미노산 중 선택되는 어느 하나 이상과 설포라판을 함유하는 혼합물에 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물을 처리함으로써, 설포라판을 장시간 안정적으로 보존할 수 있었다. In addition, the present invention was able to stably preserve sulforaphane for a long time by treating a compound containing an isothiocynate functional group in a mixture containing any one or more selected from proteins, peptides and amino acids and sulforaphane.
도 1은 미로시네이즈 (myrosinase)의 효소 기작에 의해 글루코라파닌(glucoraphanin)이 설포라판(sulforaphane)으로 전환되는 과정을 보여준다.
도 2는 PITC를 처리한 브로콜리 분쇄액의 원심분리 후 모습이다.
도 3은 PITC(phenylisothiocynate) 처리 여부에 따른 설포라판 생성량의 비교 실험 결과이다.1 shows a process in which glucoraphanin is converted to sulforaphane by an enzymatic mechanism of myrosinase.
Figure 2 is a view after centrifugation of the PITC-treated broccoli pulverized solution.
3 is a comparative experiment result of the amount of sulforaphane produced according to whether PITC (phenylisothiocynate) treatment.
십자화과 식물에는 통상적으로 설포라판(sulforaphane)의 기질(substrate)인 글루코라파닌(glucoraphanin)이 다량 포함되어 있다. 십자화과 식물에는 마이로시네이즈(myrosinase)라는 효소가 존재하는데, 이 효소는 글루코라파닌으로부터 설포라판의 생성을 유도한다. 그런데, 설포라핀이 생성되면, 십자화과 식물 내의 뉴클레오파일(nucleophile)이 설포라판과 화합물(sulforaphane conjugate)을 이루어 설포라판의 활성에 부정적인 영향을 준다. 뉴클레오파일은 친핵체로서, 단백질, 펩타이드 또는 아미노산들이 대표적이다.Cruciferous plants usually contain a large amount of glucoraphanin, which is a substrate of sulforaphane. Cruciferous plants contain an enzyme called myrosinase, which induces the production of sulforaphane from glucoraphanin. However, when sulforapine is produced, nucleophile in the cruciferous plant forms a compound (sulforaphane conjugate) with sulforaphane, thereby negatively affecting the activity of sulforaphane. A nucleophile is a nucleophile, and proteins, peptides, or amino acids are representative.
이를 극복하기 위하여 본 발명에서는 십자화과 식물 분쇄액에 과량의 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물을 처리하여 십자화과 식물 내의 단백질, 펩타이드 또는 아미노산들을 침전시켜 설포라판이 화합물화 되는 것을 원천적으로 막았다. In order to overcome this problem, in the present invention, protein, peptide or amino acids in the cruciferous plant are precipitated by treating the compound containing an excess of isothiocynate functional group in the crushed cruciferous plant, thereby fundamentally preventing sulforaphane from being compounded.
본 발명에서는 십자화과 식물 분쇄액에 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물을 처리한 후, 마이로시네이즈를 처리하면 순수한 설포라판만이 효소전환으로 생성되는 것을 확인하였다. 또한, 그 후 보관 중에도, 생성된 설포라판이 화합물화가 되지 않고, 순수한 설포라판으로 안정적으로 유지됨을 확인하였다.In the present invention, it was confirmed that only pure sulforaphane was produced by enzymatic conversion when the crushed cruciferous plant was treated with a compound containing an isothiocynate functional group and then treated with myrosinase. In addition, it was confirmed that the produced sulforaphane was not compounded and was stably maintained as pure sulforaphane even during storage thereafter.
이를 통해 본 발명은 십자화과 식물의 분쇄액에 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물을 처리하는 단계 (a); 상기 단계 (a) 후, 마이로시네이즈(myrosinase)를 처리하는 단계 (b);를 포함하는 것을 특징으로 하는 설포라판(sulforaphane)의 생산방법을 제공한다.Through this, the present invention comprises the steps of (a) treating a compound containing an isocyanate functional group in the crushed liquid of a cruciferous plant; After the step (a), the step (b) of treating myrosinase (myrosinase); provides a method for producing sulforaphane (sulforaphane) comprising the.
이하, 본 발명의 설포라판(sulforaphane) 생산방법에 대해 하기에서 각 단계별로 상세히 설명하고자 한다.Hereinafter, the sulforaphane production method of the present invention will be described in detail for each step below.
<단계 (a): 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물의 처리><Step (a): Treatment of a compound containing an isothiocynate functional group>
본 단계는 십자화과 식물의 분쇄액에 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물을 처리하는 과정이다. This step is a process of treating a compound containing an isothiocynate functional group in the crushed liquid of a cruciferous plant.
상기 십자화과 식물은, 일 예로 브로콜리, 콜라비, 레드비트, 양배추 중 선택되는 어느 하나일 수 있다. 상기 십자화과 식물의 분쇄액은 바람직하게 글루코라파닌(glucoraphanin)을 함유하는 것이 좋다. 글루코라파닌은 설포라판의 전구체로서, 하기 단계 (b)의 마이로시네이즈(myrosinase)에 의해 설포라판으로 전환된다. The cruciferous plant may be, for example, any one selected from broccoli, kohlrabi, red beet, and cabbage. The pulverized liquid of the cruciferous plant preferably contains glucoraphanin. Glucorapanin is a precursor of sulforaphane, and is converted to sulforaphane by myrosinase in step (b) below.
본 단계 (a)에서는 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물의 처리에 의해 십자화과 식물 분쇄액 내의 뉴클레오파일(nucleophile)이 침전되는데, 상기 클레오파일(nucleophile)은 단백질, 펩타이드 및 아미노산 등이 대표적이다. In this step (a), nucleophiles in the crushed cruciferous plant are precipitated by treatment with a compound containing an isothiocynate functional group, and the nucleophiles are proteins, peptides, amino acids, etc. This is an example.
십자화과 식물 내의 단백질, 펩타이드 또는 아미노산들은 설포라판이 존재하면 설포라판과 화합물(sulforaphane conjugate)을 이루어 설포라판의 수득에 부정적인 영향을 준다. 하지만, 본 발명에서는 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물을 처리함으로써 십자화과 식물 내의 단백질, 펩타이드 또는 아미노산들을 설포라판이 생성되기 전에 미리 침전시켜 설포라판이 화합물이 되는 것을 원천적으로 막을 수 있었던 것이다. Proteins, peptides or amino acids in cruciferous plants form a compound (sulforaphane conjugate) with sulforaphane when sulforaphane is present, and negatively affect the yield of sulforaphane. However, in the present invention, by treating a compound containing an isothiocynate functional group, proteins, peptides, or amino acids in the cruciferous plant were precipitated before sulforaphane was produced, thereby fundamentally preventing sulforaphane from becoming a compound.
본 발명의 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물은 아이소사이오시아네이트(isothiocynate) 작용기 (N=C=S)를 포함하는 것이면 어느 것이나 사용할 수 있으며, 일 예로는 PITC(phenylisothiocynate)가 있다. As the compound containing an isocyanate functional group of the present invention, any compound containing an isocyanate functional group (N=C=S) may be used, and an example is PITC (phenylisothiocynate). .
한편, 본 발명은 상기 단계 (a) 후, 원심분리하여 상등액을 수득하고, 수득한 상등액에 하기 단계 (b)의 마이로시네이즈를 처리할 수 있다. 단계 (a)를 거치게 되면, 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물에 의해 침전물이 생기게 되는데, 이것을 원심분리하여 상등액을 수득할 수 있다. 상등액을 분리하여 사용하면, 최종 제조하는 설포라판의 순도를 높일 수 있는 장점이 있다. 아이소사이오시아네이트(isothiocynate) 작용기의 예로 PITC를 사용한 도 2를 보면, 아래층부터 PITC 포함 불용성 부분, PITC 화합물, 상등액의 순서로 원심분리가 됨을 알 수 있고, 제일 위의 상등액을 수득하여 하기 단계 (b)에서 사용할 수 있는 것이다. 상등액에는 설포라판의 전구체인 글루코라파닌이 포함되어 있다. Meanwhile, in the present invention, after step (a), centrifugation is performed to obtain a supernatant, and the obtained supernatant may be treated with the myrosinase of step (b). When step (a) is performed, a precipitate is generated by a compound containing an isothiocynate functional group, and this can be centrifuged to obtain a supernatant. When the supernatant is separated and used, there is an advantage in that the purity of the sulforaphane finally produced can be increased. Referring to FIG. 2 using PITC as an example of an isocyanate functional group, it can be seen that centrifugation is performed in the order of an insoluble portion containing PITC, a PITC compound, and a supernatant from the lower layer, and the following steps ( b) can be used. The supernatant contains glucoraphanin, a precursor of sulforaphane.
<단계 (b): 마이로시네이즈(myrosinase) 처리> <Step (b): myrosinase treatment>
본 단게는 상기 단계 (a) 후, 마이로시네이즈(myrosinase)를 처리하는 과정이다. This step is a process of treating myrosinase after step (a).
본 단계는 설포라판을 생산하는 과정이다. 마이로시네이즈(myrosinase)는 도 1에서 보는 바와 같이 글루코라파닌으로부터 설포라판의 생산을 유도하는 효소이다. 본 단계를 거침으로써 최종적으로 설포라판이 생성된다. This step is a process for producing sulforaphane. Myrosinase (myrosinase) is an enzyme that induces the production of sulforaphane from glucoraphanin as shown in FIG. Sulforaphane is finally produced by going through this step.
한편, 본 발명은 단백질, 펩타이드 및 아미노산 중 선택되는 어느 하나 이상과 설포라판을 함유하는 혼합물에 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물를 처리하는 것을 특징으로 하는 설포라판의 보존방법을 제공한다. On the other hand, the present invention provides a method for preserving sulforaphane, comprising treating a compound containing an isothiocynate functional group to a mixture containing any one or more selected from proteins, peptides and amino acids and sulforaphane.
설포라판은 보관 중, 같이 존재하는 단백질, 펩타이드 및 아미노산 등의 뉴클레오파일에 의해 화합물화가 되어 순도가 떨어지게 되는데, 본 발명의 방법에 의할 경우, 화합물화가 되는 것을 방지하여 높은 순도를 유지할 수 있는 것이다. During storage, sulforaphane is compounded by nucleophiles such as proteins, peptides, and amino acids that are present together and the purity is lowered. .
이하, 본 발명의 내용을 하기 실시예를 통해 더욱 상세히 설명하고자 한다. 다만, 본 발명의 권리범위가 하기 실시예에만 한정되는 것은 아니고, 그와 등가의 기술적 사상의 변형까지를 포함한다.Hereinafter, the content of the present invention will be described in more detail through the following examples. However, the scope of the present invention is not limited only to the following examples, and includes modifications of technical ideas equivalent thereto.
[실시예 1: 페닐이소티오시아네이트(PITC)를 처리하여 십자화과 식물인 브로콜리로부터 설포라판 생산] [Example 1: Production of sulforaphane from broccoli, a cruciferous plant, by treatment with phenylisothiocyanate (PITC)]
본 실시예에서는 아이소사이오시아네이트(isothiocynate) 작용기를 포함하는 화합물의 일 예로서, PITC(phenylisothiocynate)를 사용하여 십자화과 식물인 브로콜리로부터 설포라판을 생산하고자 하였다. In this example, as an example of a compound containing an isothiocynate functional group, phenylisothiocynate (PITC) was used to produce sulforaphane from broccoli, a cruciferous plant.
실험을 위해, 100℃에서 20분간 열처리한 브로콜리를 분쇄하여 얻은 분쇄액에 PITC(phenylisothiocynate)를 100g/L 처리하였다. 이후, 37℃에서 10,000 rpm으로 2시간 동안 원심분리하고 상등액을 수득하였다. 수득한 상등액에 마이로시네이즈(myrosinase)를 5% 처리한 후, 40℃에서 2시간 동안 반응을 유도하였다. 이후, 반응액 샘플 (D4) 내에 존재하는 설포라판의 함량을 정량하였다. For the experiment, 100 g/L of PITC (phenylisothiocynate) was treated in the pulverized solution obtained by pulverizing broccoli heat-treated at 100° C. for 20 minutes. Thereafter, centrifugation was performed at 10,000 rpm at 37° C. for 2 hours to obtain a supernatant. After the obtained supernatant was treated with 5% myrosinase, a reaction was induced at 40° C. for 2 hours. Thereafter, the content of sulforaphane present in the reaction solution sample (D4) was quantified.
한편, 대조군으로는 브로콜리 분쇄액을 40℃에서 2시간 동안 처리한 샘플 (D1), 브로콜리 분쇄액에 마이로시네이즈를 5% 처리한 후, 40℃에서 2시간 동안 처리한 샘플 (D2), 브로콜리 분쇄액에 PITC를 처리한 후, 40℃에서 2시간 동안 처리한 샘플 (D3)을 사용하였다. 실험군은 브로콜리 분쇄액에 PITC를 처리한 후, 마이로시네이즈를 5% 처리한 후, 40℃에서 2시간 동안 처리한 샘플 (D4)이다. On the other hand, as a control, a sample treated with crushed broccoli at 40° C. for 2 hours (D1), a sample treated with 5% myrosinase in crushed broccoli solution at 40° C. for 2 hours (D2), After PITC was treated with the crushed broccoli solution, a sample (D3) treated at 40° C. for 2 hours was used. The experimental group is a sample (D4) treated with PITC in broccoli ground solution, 5% myrosinase, and then treated at 40° C. for 2 hours.
실험 결과는 도 3과 같이 나타났다. 도 3은 PITC 처리 여부에 따른 설포라판 생성량의 비교 실험 결과이다. 도 3에서 보는 바와 같이 본 발명의 샘플 D4는 PITC를 처리하지 않은 대조군 샘플 D2에 비해 설포라판의 함량이 5.7% 향상됨 {(925-875)/875*100}을 확인할 수 있었다. The experimental results were shown in FIG. 3 . 3 is a comparison experimental result of the amount of sulforaphane produced according to whether or not PITC treatment. As shown in FIG. 3 , it was confirmed that the sulforaphane content of the sample D4 of the present invention was improved by 5.7% {(925-875)/875*100} compared to the control sample D2 that was not treated with PITC.
Claims (6)
상기 단계 (a) 후, 원심분리하여 상등액을 회수한 후, 회수한 상등액에 마이로시네이즈(myrosinase)를 처리하는 단계 (b);를 포함하는 것을 특징으로 하는 고순도 설포라판(sulforaphane)의 생산방법.
Treating the crushed liquid of the cruciferous plant with phenyl isothiocyanate (phenylisothiocyanate) (a);
After the step (a), the supernatant is recovered by centrifugation, and the recovered supernatant is treated with myrosinase (b); .
상기 십자화과 식물은,
브로콜리, 콜라비, 레드비트, 양배추 중 선택되는 어느 하나인 것을 특징으로 하는 고순도 설포라판(sulforaphane)의 생산방법.
The method of claim 1,
The cruciferous plant,
A method of producing high purity sulforaphane, characterized in that any one selected from broccoli, kohlrabi, red beet, and cabbage.
상기 십자화과 식물의 분쇄액에는,
글루코라파닌(glucoraphanin)이 함유되어 있는 것을 특징으로 하는 고순도 설포라판(sulforaphane)의 생산방법.
The method of claim 1,
In the crushed liquid of the cruciferous plant,
A method of producing high-purity sulforaphane, characterized in that it contains glucoraphanin.
상기 단계 (a)는,
페닐이소티오시아네이트(phenylisothiocyanate)의 처리에 의해 십자화과 식물 분쇄액 내의 단백질, 펩타이드 및 아미노산 중 선택되는 어느 하나 이상이 침전되는 것을 특징으로 하는 고순도 설포라판(sulforaphane)의 생산방법.
The method of claim 1,
The step (a) is,
A method for producing high-purity sulforaphane, characterized in that at least one selected from proteins, peptides and amino acids in the crushed cruciferous plant is precipitated by treatment with phenylisothiocyanate.
A high-purity preservation method of sulforaphane, characterized in that a mixture containing at least one selected from protein, peptide and amino acid and sulforaphane is treated with phenylisothiocyanate.
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| JP2010516766A (en) | 2007-01-23 | 2010-05-20 | フアーマグラ・ラブズ・インコーポレイテツド | Stabilized sulforaphane |
| JP2012520063A (en) | 2009-03-12 | 2012-09-06 | ネステク ソシエテ アノニム | Milk protein / isothiocyanate covalent complex |
| JP2014506597A (en) | 2011-02-22 | 2014-03-17 | コーディル・シード・カンパニー・インコーポレイテッド | Spray dried myrosinase and use for producing isothiocyanate |
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| KR100946727B1 (en) | 2008-03-18 | 2010-03-12 | 한국과학기술연구원 | How to amplify the sulforaphane content of broccoli through ultra high pressure treatment |
| KR101761575B1 (en) | 2015-09-08 | 2017-07-26 | (주)제주아침 | Method for Manufacturing Broccoli Juice Having High Sulforaphane Content and Method for Manufacturing Bean Curd Using the Juice |
| KR101965391B1 (en) | 2018-04-17 | 2019-04-03 | 주태영 | Broccoli Sprout Extract with Enhanced Sulforaphane and Manufacturing Method Thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2010516766A (en) | 2007-01-23 | 2010-05-20 | フアーマグラ・ラブズ・インコーポレイテツド | Stabilized sulforaphane |
| JP2012520063A (en) | 2009-03-12 | 2012-09-06 | ネステク ソシエテ アノニム | Milk protein / isothiocyanate covalent complex |
| JP2014506597A (en) | 2011-02-22 | 2014-03-17 | コーディル・シード・カンパニー・インコーポレイテッド | Spray dried myrosinase and use for producing isothiocyanate |
Non-Patent Citations (1)
| Title |
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| Food Chemistry, Vol. 268, pp. 249-256 (2018.06.19.)* |
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