KR102418178B1 - Method of manufacturing Gryllus bimaculatus protein extract and powder - Google Patents

Method of manufacturing Gryllus bimaculatus protein extract and powder Download PDF

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KR102418178B1
KR102418178B1 KR1020190134349A KR20190134349A KR102418178B1 KR 102418178 B1 KR102418178 B1 KR 102418178B1 KR 1020190134349 A KR1020190134349 A KR 1020190134349A KR 20190134349 A KR20190134349 A KR 20190134349A KR 102418178 B1 KR102418178 B1 KR 102418178B1
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cricket
protein
star
crickets
powder
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김용욱
박보람
류정표
장현욱
최지호
박신영
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주식회사 케일
대한민국(농촌진흥청장)
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00—Working-up of proteins for foodstuffs
    • A23J3/04—Animal proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00—Working-up of proteins for foodstuffs
    • A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L35/00—Food or foodstuffs not provided for in groups A23L5/00 – A23L33/00; Preparation or treatment thereof

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Abstract

본 발명은 쌍별귀뚜라미 원료의 지질이 분리되어 산패가 억제되고, 단백질 가수분해효소 처리를 통해 고수율의 수용성 아미노산을 추출할 수 있는 구성이 우수한 쌍별귀뚜라미 단백질 추출물 및 분말을 제조하는 방법을 제공하며, 이를 통해 제조된 쌍별귀뚜라미 단백질 추출물 및 분말은 저장안정성 및 품질이 우수하여 다양한 식품 조성물에 적용이 가능하다.The present invention provides a method for producing a bipolar cricket protein extract and powder having an excellent composition in which the lipids of the raw material of crickets are separated, rancidity is suppressed, and a high yield of water-soluble amino acids can be extracted through proteolytic enzyme treatment, The bipolar cricket protein extract and powder prepared through this process have excellent storage stability and quality, so they can be applied to various food compositions.

Description

쌍별귀뚜라미 단백질 추출물 및 분말의 제조방법{Method of manufacturing Gryllus bimaculatus protein extract and powder}Method of manufacturing Gryllus bimaculatus protein extract and powder

쌍별귀뚜라미 단백질 추출물 및 분말과, 이의 제조방법에 관한 것이다.It relates to a cricket protein extract and powder, and a method for preparing the same.

국제식량농업기구(FAO)에서는 곤충을 미래 대체 식량으로 주목하고 있으며, 우리나라에서는 곤충시장 확대 및 농가 소득원 창출을 위해 곤충이 식품 원료로 이용되는 것이 시급했고, 한시적 인정 요청 제도 신설로 식용곤충을 도입하면서 새로운 식품 인증 절차를 완화하게 되었다. 한국의 경우 귀뚜라미, 메뚜기, 번데기, 누에가 식약청 승인을 받아 일반식품으로 사용되고 있는데, 이러한 식용 곤충을 먹는 나라로는 중국, 태국, 일본, 남아프리카 공화국, 멕시코, 잠비아 등 전 세계 20억 명에 달하는 인구가 대략 1900종의 식용 곤충을 음식으로 먹는다고 한다. 또한, 2003년 이후 유엔농업식량기구(FAO)는 전세계 국가와 긴밀하게 협의하며 식용 곤충식의 확대와 개발 필요성에 대해 끊임없이 홍보하고 있으며, 네덜란드, 영국, 미국, 일본 등 많은 나라들이 식용 곤충식 활성화에 참여하고 있다. 특히, CNN, TIME지 등의 보도매체를 통해 그간 식용 곤충에 부정적이었던 서구권에서도 식용 곤충 식용화에 눈을 돌리고 있다.The Food and Agriculture Organization (FAO) is paying attention to insects as a future alternative food. The new food certification process was relaxed. In Korea, crickets, grasshoppers, pupae, and silkworms have been approved by the Food and Drug Administration and are used as general food. Countries that eat these edible insects include China, Thailand, Japan, South Africa, Mexico and Zambia, with a population of 2 billion people worldwide. It is said that they eat about 1900 species of edible insects as food. In addition, the Food and Agriculture Organization of the United Nations (FAO) has been continuously promoting the need for expansion and development of edible insect food in close consultation with countries around the world since 2003. is participating in In particular, through news media such as CNN and TIME magazine, Western countries, which have been negative about edible insects, are turning their attention to edible insects.

식용곤충으로서 미래의 단백질원으로 주목받고 있는 곤충 중 하나인 쌍별귀뚜라미는 단백질 함량이 매우 높고 구성이 우수하여 이용가치가 높다. 쌍별귀뚜라미에 관해 쌍별귀뚜라미의 사육밀도가 섭식량, 성충사망률 및 부화 약충수에 미치는 영향(박영규 외, 2013. 한국잠사곤충학회지 51, 89-94)이 보고되어 있으며, 사료 및 식용화를 위해 귀뚜라미 첨가 사료가 계육과 계란의 성분에 미치는 영향(안미영 외, 2000. 한국가금학회지 27, 197-202), 산지별 식용 귀뚜라미의 이화학적 특성(김은미 외, 2015. 한국식품저장유통학회지, 22, 831-837) 및 랫드에서 귀뚜라미 추출물을 반복투여한 경우 독성이 없음이 보고(황석연 외, 2004. 한국실험동물학회지 20, 194-199)되어 있다. 그러나, 쌍별귀뚜라미는 지방성분이 함유되어 있어 변질의 위험이 존재하는 문제점이 있고, 단백질 성분의 가용화 처리 등의 농축소재 제조공정의 개발이 필요한 실정이다.As an edible insect, one of the insects that is attracting attention as a protein source in the future, the twin-star cricket has a very high protein content and excellent composition, so its use value is high. Regarding twin-star crickets, the effect of breeding density of twin-star crickets on feeding amount, adult mortality rate and the number of hatching nymphs (Park Young-gyu et al., 2013. Journal of the Korean Society of Sleeping Insects 51, 89-94) has been reported. Effect on ingredients of domestic chicken meat and eggs (Miyoung Ahn et al., 2000. Journal of the Korean Poultry Society 27, 197-202), Physicochemical characteristics of edible crickets by production area (Eunmi Kim et al., 2015. Journal of the Korean Society for Food Storage and Distribution, 22, 831-837) ) and no toxicity in the case of repeated administration of cricket extract in rats (Hwang Seok-yeon et al., 2004. Journal of the Korean Society of Laboratory Animals 20, 194-199). However, there is a problem in that there is a risk of deterioration due to the presence of fat components in pair-star crickets, and it is necessary to develop a process for manufacturing a concentrated material such as solubilization treatment of protein components.

본 발명의 일 목적은 지질이 분리되고 단백질 함량이 높은 쌍별귀뚜라미 단백질 추출물 및 분말의 제조방법을 제공하는 것이다.It is an object of the present invention to provide a method for preparing a bipolar cricket protein extract and powder in which lipids are separated and protein content is high.

본 발명의 다른 일 측면은 상기 쌍별귀뚜라미 단백질 추출물 및 분말을 함유하는 식품 조성물을 제공하는 것이다.Another aspect of the present invention is to provide a food composition containing the bipolar cricket protein extract and powder.

상기 목적을 달성하기 위하여,In order to achieve the above object,

본 발명은, 착유기로 투입하는 쌍별귀뚜라미의 투입온도를 70℃ 내지 100℃로 만드는 투입온도 설정 단계;The present invention, the input temperature setting step of making the input temperature of the pair of crickets put into the milking machine from 70 ℃ to 100 ℃;

상기 투입온도의 쌍별귀뚜라미를 60℃ 내지 100℃의 온도에서 압착하여 착유하는 착유단계; 및A milking step of pressing and milking the paired crickets at the input temperature at a temperature of 60°C to 100°C; and

상기 착유된 쌍별귀뚜라미를 플레버자임(Flavourzyme), 프로타맥스(Protamax), 알칼레이즈(Alcalase) 및 뉴트라제(Neutrase)로 이루어진 군에서 선택된 1종 이상의 가수분해효소를 이용하여 추출하는 추출단계;를 포함하는 쌍별귀뚜라미 단백질 추출물의 제조방법을 제공한다.Extraction step of extracting the milked twin-star crickets using one or more hydrolytic enzymes selected from the group consisting of Flavorzyme, Protamax, Alcalase and Neutrase It provides a method for producing a bipolar cricket protein extract comprising;

또한, 본 발명은, 착유기로 투입하는 쌍별귀뚜라미의 투입온도를 70℃ 내지 100℃로 만드는 투입온도 설정 단계;In addition, the present invention, the input temperature setting step of making the input temperature of the pair of crickets put into the milking machine from 70 ℃ to 100 ℃;

상기 투입온도의 쌍별귀뚜라미를 60℃ 내지 100℃의 온도에서 압착하여 착유하는 착유단계;A milking step of pressing and milking the paired crickets at the input temperature at a temperature of 60°C to 100°C;

상기 착유된 쌍별귀뚜라미를 플레버자임(Flavourzyme), 프로타맥스(Protamax), 알칼레이즈(Alcalase) 및 뉴트라제(Neutrase)로 이루어진 군에서 선택된 1종 이상의 가수분해효소를 이용하여 추출하는 추출단계;Extraction step of extracting the milked twin-star crickets using one or more hydrolytic enzymes selected from the group consisting of Flavorzyme, Protamax, Alcalase and Neutrase ;

상기 추출된 추출물을 50℃ 내지 60℃에서 감압농축하는 농축단계; 및a concentration step of concentrating the extracted extract under reduced pressure at 50°C to 60°C; and

상기 농축된 추출물을 분무건조하는 하여 분말을 제조하는 분말화단계;를 포함하는 쌍별귀뚜라미 단백질 분말의 제조방법을 제공한다.It provides a method for producing a bi-star cricket protein powder comprising; a powdering step of preparing a powder by spray-drying the concentrated extract.

또한, 본 발명은 상기 쌍별귀뚜라미 단백질 추출물의 제조방법으로 제조된 쌍별귀뚜라미 단백질 추출물을 함유하는 식품 조성물을 제공하고, 및In addition, the present invention provides a food composition containing the bipolar cricket protein extract prepared by the method for producing the bipolar cricket protein extract, and

상기 쌍별귀뚜라미 단백질 분말의 제조방으로 제조된 쌍별귀뚜라미 단백질 분발을 함유하는 식품 조성물을 제공한다.It provides a food composition containing the pair-star cricket protein powder prepared by the production room of the pair-star cricket protein powder.

본 발명의 쌍별귀뚜라미 단백질 추출물 및 분말은 지질이 분리되어 산패가 억제되고, 단백질 가수분해효소 처리를 통해 고수율의 수용성 아미노산을 추출할 수 있어서 구성이 우수하고, 저장안정성 및 품질확보를 통해 다양한 식품 조성물에 적용이 가능하다.The pairwise cricket protein extract and powder of the present invention are excellent in composition because lipids are separated to suppress rancidity, and high yield of water-soluble amino acids can be extracted through proteolytic enzyme treatment, and various food products through storage stability and quality assurance It can be applied to the composition.

도 1은 쌍별귀뚜라미 단백질 추출액 및 분말의 제조공정을 나타낸 것이다.
도 2는 쌍별귀뚜라미 단백질 추출액의 제조공정별로 쌍별귀뚜라미 건조 원료, 쌍별귀뚜라미 착유박, 단백질 가수분해 효소를 처리한 쌍별귀뚜라미 단백농축액, 쌍별귀뚜라미 단백농축분말의 대사체 함량 변화를 분석한 결과를 나타낸 것이다.
도 3은 쌍별귀뚜라미 건조 원료, 쌍별귀뚜라미 착유박, 단백가수분해 효소를 처리한 쌍별귀뚜라미 단백농축액, 쌍별귀뚜라미 단백농축분말 대사체를 크로마토그래피로 분석한 결과를 나타낸 것이다.1 shows the manufacturing process of the bipolar cricket protein extract and powder.
Figure 2 shows the results of analyzing the metabolite content change of the pairwise cricket dry raw material, the pairwise cricket milk meal, the pairwise cricket protein concentrate treated with proteolytic enzymes, and the pairwise cricket protein concentrate powder for each manufacturing process of the pairwise cricket protein extract. .
3 shows the results of chromatographic analysis of twin-star cricket dry raw materials, twin-star cricket milk meal, protein hydrolytic enzyme-treated twin-star cricket protein concentrate, and twin-star cricket protein concentrate powder metabolites.

이하, 본 발명을 상세하게 설명한다.Hereinafter, the present invention will be described in detail.

본 발명은, 착유기로 투입하는 쌍별귀뚜라미의 투입온도를 70℃ 내지 100℃로 만드는 투입온도 설정 단계;The present invention, the input temperature setting step of making the input temperature of the pair of crickets put into the milking machine from 70 ℃ to 100 ℃;

상기 투입온도의 쌍별귀뚜라미를 60℃ 내지 100℃의 온도에서 압착하여 착유하는 착유단계; 및A milking step of pressing and milking the paired crickets at the input temperature at a temperature of 60°C to 100°C; and

상기 착유된 쌍별귀뚜라미를 플레버자임(Flavourzyme), 프로타맥스(Protamax), 알칼레이즈(Alcalase) 및 뉴트라제(Neutrase)로 이루어진 군에서 선택된 1종 이상의 가수분해효소를 이용하여 추출하는 추출단계;를 포함하는 쌍별귀뚜라미 단백질 추출물의 제조방법을 제공한다.Extraction step of extracting the milked twin-star crickets using one or more hydrolytic enzymes selected from the group consisting of Flavorzyme, Protamax, Alcalase and Neutrase It provides a method for producing a bipolar cricket protein extract comprising;

상기 쌍별귀뚜라미의 투입온도는 착유시의 온도(예를들면 착유기의 온도)와는 별개로, 착유의 대상이 되는 쌍별귀뚜라미 원료의 온도를 설정한 것이다. 예를들어, 쌍별귀뚜라미를 가열 등을 통해 온도를 70℃ 내지 100℃로 만든 다음, 착유기에 투입할 수 있다.The input temperature of the paired crickets is set to the temperature of the raw material of the paired crickets to be milked separately from the temperature at the time of milking (eg, the temperature of a milking machine). For example, the pair-star crickets may be heated to 70° C. to 100° C. and then put into the milking machine.

상기 쌍별귀뚜라미의 투입온도는 75℃ 내지 85℃일 수 있고, 90℃ 또는 100℃일 수 있고, 바람직하게는 70℃일 수 있고, 가장 바람직하게는 80℃의 투입온도로 만들 수 있다.The input temperature of the pair-star crickets may be 75°C to 85°C, 90°C or 100°C, preferably 70°C, and most preferably 80°C.

상기 착유단계에서 쌍별귀뚜라미를 65℃ 내지 75℃에서 착유할 수 있는데, 바람직하게는 70℃에서 착유할 수 있다. 이때의 온도는 상기 투입온도(원료 온도)와는 별개로 착유를 실시할 때의 온도, 예를 들면 착유기의 온도를 말하는 것이다.In the milking step, the paired crickets may be milked at 65°C to 75°C, preferably at 70°C. The temperature at this time refers to the temperature at the time of milking separately from the input temperature (raw material temperature), for example, the temperature of the milking machine.

상기 추출단계에서 이용하는 가수분해효소의 총 농도는 0.10w/w% 내지 0.50w/w%일 수 있고, 0.15w/w% 내지 0.40w/w%일 수 있고, 0.20w/w% 내지 0.35w/w%일 수 있고, 바람직하게는 0.20w/w%일 수 있다.The total concentration of the hydrolase used in the extraction step may be 0.10w/w% to 0.50w/w%, 0.15w/w% to 0.40w/w%, 0.20w/w% to 0.35w It may be /w%, preferably 0.20w/w%.

상기 가수분해효소의 처리는 플레버자임(Flavourzyme), 프로타맥스(Protamax), 알칼레이즈(Alcalase) 및 뉴트라제(Neutrase)를 순서를 조합하여 처리할 수 있는데, 예를 들어 뉴트라제, 알칼레이즈, 프로타맥스, 플레버자임 순으로 처리할 수도 있고, 알칼레이즈, 프로타맥스, 뉴트라제, 플레버자임 순으로 처리할수도 있는데, 가장 바람직하게는 플레버자임, 프로타맥스, 알칼레이즈, 뉴트라제 순으로 처리하는 것이다.The treatment of the hydrolase can be treated by combining the sequence of Flavorzyme, Protamax, Alcalase and Neutrase, for example, Neutrase, Al Callase, Protamax, Flavorzyme may be processed in the order, Alkalease, Protamax, Neutrase, Flavorzyme may be processed in the order, most preferably, Flavorzyme, Protamax, Alkalease and neutrase are treated in that order.

상기 상기 추출단계의 가수분해 효소 처리 시간은 40분 내지 100분 동안 처리할 수 있는데, 50분 내지 90분 동안 처리할 수도 있고, 60분 내지 90분 동안 처리할 수도 있고, 바람직하게는 60분 또는 90분 동안 처리할 수 있다.The hydrolase treatment time of the extraction step may be treated for 40 minutes to 100 minutes, may be treated for 50 minutes to 90 minutes, may be treated for 60 minutes to 90 minutes, preferably 60 minutes or It can be processed for 90 minutes.

본 발명은, 착유기로 투입하는 쌍별귀뚜라미의 투입온도를 70℃ 내지 100℃로 만드는 투입온도 설정 단계;The present invention, the input temperature setting step of making the input temperature of the pair of crickets put into the milking machine from 70 ℃ to 100 ℃;

상기 투입온도의 쌍별귀뚜라미를 60℃ 내지 100℃의 온도에서 압착하여 착유하는 착유단계;A milking step of pressing and milking the paired crickets at the input temperature at a temperature of 60°C to 100°C;

상기 착유된 쌍별귀뚜라미를 플레버자임(Flavourzyme), 프로타맥스(Protamax), 알칼레이즈(Alcalase) 및 뉴트라제(Neutrase)로 이루어진 군에서 선택된 1종 이상의 가수분해효소를 이용하여 추출하는 추출단계;Extraction step of extracting the milked twin-star crickets using one or more hydrolytic enzymes selected from the group consisting of Flavorzyme, Protamax, Alcalase and Neutrase ;

상기 추출된 추출물을 50℃ 내지 60℃에서 감압농축하는 농축단계; 및a concentration step of concentrating the extracted extract under reduced pressure at 50°C to 60°C; and

상기 농축된 추출물을 분무건조하는 하여 분말을 제조하는 분말화단계;를 포함하는 쌍별귀뚜라미 단백질 분말의 제조방법을 제공한다.It provides a method for producing a bi-star cricket protein powder comprising; a powdering step of preparing a powder by spray-drying the concentrated extract.

이때 상기 투입온도 설정단계, 착유단계, 추출단계의 조건은 상기 쌍별귀뚜라미 단백질 추출물의 제조방법에서 서술한 바와 같다.At this time, the conditions of the input temperature setting step, the milking step, and the extraction step are the same as those described in the method for preparing the cricket protein extract by pair.

본 발명은 상기 쌍별귀뚜라미 단백질 추출물의 제조방법으로 제조한 쌍별귀뚜라미 단백질 추출물을 함유하는 식품조성물을 제공한다.The present invention provides a food composition containing the bipolar cricket protein extract prepared by the method for producing the bipolar cricket protein extract.

본 발명은 상기 쌍별귀뚜라미 단백질 분말의 제조방법으로 제조한 쌍별귀뚜라미 단백질 분말을 함유하는 식품조성물을 제공한다.The present invention provides a food composition containing the pair-star cricket protein powder prepared by the method for producing the pair-star cricket protein powder.

상기 본 발명에 따른 식품 조성물은 일반 식품, 기능성 식품(functional food), 영양 보조제(nutritional supplement), 건강식품(health food) 및 식품 첨가제(food additives) 등의 모든 형태를 포함한다. 상기 유형의 식품 조성물은 당 업계에 공지된 통상적인 방법에 따라 다양한 형태로 제조할 수 있다.The food composition according to the present invention includes all types of general food, functional food, nutritional supplement, health food, and food additives. Food compositions of this type can be prepared in various forms according to conventional methods known in the art.

예를 들면, 건강식품으로는 본 발명의 쌍별귀뚜라미 추출물 또는 분말, 및 유산균 또는 상기 유산균의 배양액의 혼합물 자체를 차, 주스 및 드링크의 형태로 제조하여 음용하도록 하거나, 페이스트화, 과립화, 캡슐화, 정제화 및 분말화하여 섭취할 수 있다. 또한, 상기 쌍별귀뚜라미 추출물 또는 분말, 및 유산균 또는 상기 유산균의 배양액의 혼합물은 분말 또는 농축액 형태로 제조하여 유소아 및 청소년 성장촉진을 목적으로 하는 식품에 첨가할 수 있다. 예를 들면, 음료, 과실 및 그의 가공식품(예: 과일통조림, 병조림, 잼, 마아말레이드 등), 어류, 육류 및 그 가공 식품(예: 햄, 소시지 콘비이프 등), 빵류 및 면류(예: 우동, 메밀국수, 라면, 스파게티, 마카로니 등), 과즙, 각종 드링크, 쿠키, 유제품(예: 버터, 치즈 등), 식용식물유지, 마아가린, 식물성 단백질, 레토르트 식품, 냉동식품, 각종 조미료(예: 된장, 간장, 소스 등) 등에 첨가할 수 있다.For example, as a health food, the extract or powder of the bipolar cricket of the present invention, and the lactic acid bacteria or a mixture of the culture solution of the lactic acid bacteria itself are prepared in the form of tea, juice, and drink to drink, paste, granulation, encapsulation, It can be ingested by tableting and powdering. In addition, the pairwise cricket extract or powder, and the lactic acid bacteria or a mixture of the culture solution of the lactic acid bacteria can be prepared in the form of a powder or a concentrate and added to food for the purpose of promoting growth in children and adolescents. For example, beverages, fruits and their processed foods (e.g. canned fruit, canned fruit, jam, marmalade, etc.), fish, meat and their processed foods (e.g. ham, sausage corned beef, etc.), breads and noodles (e.g. : Udon, soba, ramen, spaghetti, macaroni, etc.), fruit juice, various drinks, cookies, dairy products (eg butter, cheese, etc.), edible vegetable oil, margarine, vegetable protein, retort food, frozen food, various seasonings (eg, butter, cheese, etc.) : Doenjang, soy sauce, sauce, etc.) can be added.

본 발명의 식품 조성물 중 쌍별귀뚜라미 추출물 또는 분말, 및 유산균 또는 상기 유산균의 배양액의 혼합물의 함량은 특별히 제한되는 것은 아니지만, 식품의 전체 중량을 기준으로 0.01~90%, 바람직하게는 0.1~50%의 비율로 함유될 수 있다. 나아가, 본 발명의 식품 조성물은 쌍별귀뚜라미 추출물 또는 분말 또는 유산균과 쌍별귀뚜라미 혼합물 성분 이외에 미량의 미네랄, 비타민, 지질, 당류 및 공지의 면역 활성을 가진 성분 등을 추가로 함유할 수 있다. 상기 미네랄로는 칼슘, 철 등 성장기에 필요한 영양성분이 함유될 수 있으며 비타민으로는 비타민 C, 비타민 E, 비타민 B1, 비타민 B2, 비타민 B6 등이 함유될 수 있다. 지질로는 알콕시글리세롤 또는 레시틴 등이 함유될 수 있으며 당류로는 프락토올리고당 등이 함유될 수 있다.In the food composition of the present invention, the content of the locust cricket extract or powder, and the lactic acid bacteria or the mixture of the lactic acid bacteria culture medium is not particularly limited, but 0.01 to 90%, preferably 0.1 to 50% based on the total weight of the food. may be contained in a proportion. Furthermore, the food composition of the present invention may further contain trace amounts of minerals, vitamins, lipids, sugars, and components having known immune activity, in addition to the components of the cricket extract or powder or the mixture of lactic acid bacteria and the crickets. The minerals may contain nutrients necessary for growth, such as calcium and iron, and vitamins may include vitamin C, vitamin E, vitamin B1, vitamin B2, vitamin B6, and the like. The lipid may include alkoxyglycerol or lecithin, and the saccharide may include fructooligosaccharide.

본 발명에 따른 쌍별귀뚜라미 추출물 또는 분말, 및 유산균 또는 상기 유산균의 배양액의 혼합물을 식품, 음료 등의 건강보조 또는 건강기능성 식품에 첨가할 수 있다. 이 경우, 쌍별귀뚜라미 추출물 또는 분말, 및 유산균 또는 상기 유산균의 배양액의 혼합물을 식품 첨가물로 사용시에 원료에 대하여 0.01 내지 20 중량%, 바람직하게는 0.1 내지 5 중량%의 양으로 첨가할 수 있다. 유효 성분의 혼합양은 사용목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. 그러나, 건강 및 위생을 목적으로 하거나 건강 조절을 목적으로 하는 장기간의 섭취의 경우에 상기 양은 상기 범위 이하일 수 있으며, 안정성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 시용될 수 있다. 상기 쌍별귀뚜라미 추출물 또는 분말, 및 유산균 또는 상기 유산균의 배양액의 혼합물을 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다.The pairwise cricket extract or powder according to the present invention, and lactic acid bacteria or a mixture of the lactic acid bacteria culture solution may be added to health supplements or health functional foods such as food and beverages. In this case, it is possible to add in an amount of 0.01 to 20% by weight, preferably 0.1 to 5% by weight, based on the raw material when using the mixture of the locust cricket extract or powder, and the lactic acid bacteria or the culture solution of the lactic acid bacteria as a food additive. The mixing amount of the active ingredient may be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment). However, in the case of long-term ingestion for health and hygiene purposes or health control, the above amount may be less than the above range, and since there is no problem in terms of stability, the active ingredient may be applied in an amount above the above range . The pairwise cricket extract or powder, and the lactic acid bacteria or a mixture of the lactic acid bacteria culture solution may be used together with other foods or food ingredients, and may be appropriately used according to a conventional method.

상기 식품의 종류에는 특별한 제한이 없다. 상기 추출물을 첨가할 수 있는 식품의 예로는 육류, 소시지, 빵, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함하는 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알코올 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다.There is no particular limitation on the type of the food. Examples of foods to which the extract can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks , alcoholic beverages and vitamin complexes, and includes all health foods in the ordinary sense.

본 발명의 식품 조성물은 여러 가지 향미제 또는 천연 탄수화물 등을 사용할 수 있다. 상기 천연 탄수화물은 포도당, 과당과 같은 단당류, 말토오스, 수크로오스 등과 같은 이당류 및 덱스트린, 시클로덱스트린과 같은 다당류, 자일리톨, 솔비톨, 에르트리톨 등의 당알코올이다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연감미료나, 사카린, 아스파르탐과 같은 합성감미료 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명에 따른 조성물 100 중량부당 일반적으로 약 0.01 내지 0.04 중량부, 바람직하게는 0.02 내지 0.03 중량부이다.In the food composition of the present invention, various flavoring agents or natural carbohydrates may be used. The natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and ertritol. As the sweetener, natural sweeteners such as tau martin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 parts by weight, preferably 0.02 to 0.03 parts by weight, per 100 parts by weight of the composition according to the present invention.

상기 외에 본 발명에 따른 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 중점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 본 발명에 따른 조성물은 천연 과일 주스 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있으며, 첨가제의 비율은 크게 중요하진 않지만 본 발명에 따른 조성물 100 중량부당 0.01 내지 0.1 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the composition according to the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal agents, pH regulators, stabilizers, preservatives, glycerin, It may contain alcohol, a carbonation agent used in carbonated beverages, and the like. In addition, the composition according to the present invention may contain pulp for the production of natural fruit juices and vegetable beverages. These components may be used independently or in combination, and the proportion of the additives is not particularly important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition according to the present invention.

이하, 본 발명의 실시예 및 실험예를 하기에 구체적으로 예시하여 설명한다. 다만, 후술하는 실시예 및 실험예는 본 발명의 일부를 예시하는 것일 뿐, 본 발명에 이에 한정되는 것은 아니다.Hereinafter, Examples and Experimental Examples of the present invention will be specifically illustrated and described below. However, the Examples and Experimental Examples to be described below are only illustrative of a part of the present invention, and are not limited thereto.

쌍별귀뚜라미 원료의 준비Preparation of raw materials for twin-star crickets

식용곤충으로 사용되는 쌍별귀뚜라미와 장수풍뎅이 유충의 수분함량, 조지방(crude fat), 조단백질(crude protein), 조회분(crude ash) 및 조섬유(crude fiber)의 함량을 분석하여 그 평균값을 하기 표 1에 나타내었다.The average values of the moisture content, crude fat, crude protein, crude ash and crude fiber content of twin-star cricket and long-lived beetle larvae used as edible insects were analyzed and the average values are shown in Table 1 below. shown in

조성물(%)Composition (%) 쌍별귀뚜라미double star cricket 장수풍뎅이 유충long-lived beetle larva 수분moisture 5.955.95 8.388.38 조지방george 12.8712.87 32.4532.45 조단백crude protein 67.7267.72 44.5844.58 조회분views 4.474.47 2.052.05 조섬유crude fiber 7.737.73 6.796.79

(g/100g, dry basis)(g/100g, dry basis)

상기 표 1에서 확인할 수 있듯이, 쌍별귀뚜라미 건조물의 조단백 함량은 주요 단백질 급원 식품인 육류(15.61~24%), 난류(12.04%), 어류(17.5~21.06%) 등에 비해 높은 함량을 보여 우수한 단백질 급원으로써 활용을 기대할 수 있을 것으로 확인하여 본 발명에서 식용곤충으로 이용가치가 높은 곤충으로서 쌍별귀뚜라미를 선택하였다.As can be seen in Table 1 above, the crude protein content of the dried crickets by pair was higher than that of meat (15.61~24%), eggs (12.04%), fish (17.5~21.06%), etc., which are the main protein source foods. As it was confirmed that it could be expected to be utilized as a edible insect, the twin-star cricket was selected as an insect with high utility value as an edible insect in the present invention.

본 발명에서 사용된 곤충은 쌍별귀뚜라미 성충으로 덕동분교 친환경 곤충농장 및 농업회사법인 미래도시농업, 농업회사법인 ㈜에코엠으로부터 구매하여 사용하였다. 시료는 72시간 절식 또는 절식 후 세척하여 살균한 후에 열풍건조시킨 제품을 구매하여 사용하였다.Insects used in the present invention were adult twin-star crickets, and were purchased from the Deokdong Branch School Eco-Friendly Insect Farm, Mirae Urban Agriculture, an agricultural corporation, and Eco-M, an agricultural corporation. Samples were fasted for 72 hours or washed and sterilized after fasting, and then hot air-dried products were purchased and used.

<실시예 1> 쌍별귀뚜라미의 착유 공정<Example 1> Milking process of twin-star crickets

1.1. 지방추출법 설정1.1. Fat extraction method setting

식용곤충과 같은 건조물의 지방추출법에는 유기용매를 이용한 화학적추출법과 온도, 압력을 이용한 물리적추출법이 있으나, 유기용매를 이용한 화학적추출의 경우 용매로 인한 인체유해성, 환경오염, 오폐수 관리, 고가의 설비구축 등의 이유로 가공공정을 확립하기 어려운 점이 있어, 일반적인 식물성 유지에서 많이 사용되는 압착법을 사용하여 착유하였다.Fat extraction methods for dry matter such as edible insects include chemical extraction using organic solvents and physical extraction using temperature and pressure. It is difficult to establish a processing process for the following reasons, so milking was performed using the compression method commonly used in general vegetable oils and fats.

1.2. 투입온도 및 착유기 내 온도 설정1.2. Setting the input temperature and the temperature in the milking machine

지방의 경우 열을 가하여 착유공정을 개발할 시 지방의 수율이 증가하여 작업의 효율성은 높아지지만, 가열된 온도로 인하여 지방의 산패를 유도할 수도 있고 품질이 낮아지는 결과를 가져오게 된다. 따라서 비교적 저온이라 할 수 있는 100℃ 미만의 온도에서 착유를 진행하고자 하였다. 엑스펠러(expeller) 착유기를 사용하여 착유기 온도를 70℃로 설정하였으며, 투입온도는 70℃, 80℃, 90℃, 100℃로 설정하여 투입량에 따른 수율과 처리량을 측정하였다.In the case of fat, when the milking process is developed by applying heat, the yield of fat increases and the efficiency of work increases, but the heated temperature may induce rancidity of the fat and lower the quality. Therefore, it was attempted to proceed with milking at a temperature of less than 100°C, which is a relatively low temperature. The expeller temperature was set to 70°C using an expeller milking machine, and the input temperature was set to 70°C, 80°C, 90°C, and 100°C to measure the yield and throughput according to the input amount.

실시예 1-1: 투입온도 70℃, 착유기 온도 70℃, 쌍별귀뚜라미 투입량 5 kgExample 1-1: input temperature 70 ℃, milking machine temperature 70 ℃, twin-star cricket input amount 5 kg

실시예 1-2: 투입온도 80℃, 착유기 온도 70℃, 쌍별귀뚜라미 투입량 5 kgExample 1-2: input temperature 80 ℃, milking machine temperature 70 ℃, twin-star cricket input amount 5 kg

실시예 1-3: 투입온도 90℃, 착유기 온도 70℃, 쌍별귀뚜라미 투입량 5 kgExample 1-3: input temperature 90 ℃, milking machine temperature 70 ℃, twin-star cricket input amount 5 kg

실시예 1-4: 투입온도 100℃, 착유기 온도 70℃, 쌍별귀뚜라미 투입량 5 kgExample 1-4: input temperature 100 ℃, milking machine temperature 70 ℃, twin-star cricket input amount 5 kg

그 결과, 투입온도에 따라 착유량 및 처리시간에 차이가 발생하였으나, 유의미한 차이는 없었다. 또한, 투입온도가 낮은 구간인 실시예 1-1 및 실시예 1-2에서 처리결과가 좋게 나타났다. 가장 높은 착유조건은 투입온도가 80℃인 실시예 1-3 에서 5 kg의 쌍별귀뚜라미를 착유하는데 9분이 걸렸으며, 수율도 11.84%로 가장 높게 나타났다.As a result, there was a difference in milking amount and treatment time depending on the input temperature, but there was no significant difference. In addition, the treatment results were good in Examples 1-1 and 1-2, which are sections in which the input temperature was low. The highest milking conditions were that it took 9 minutes to milk 5 kg of twin-star crickets in Example 1-3 where the input temperature was 80° C., and the yield was also the highest at 11.84%.

<실시예 2> 쌍별귀뚜라미의 단백질 가수분해를 위한 활성 효소 측정<Example 2> Measurement of active enzymes for protein hydrolysis of paired crickets

본 발명에서 사용된 상업용 효소는 노보자임(Novozymes, Bagsvard, Denmark)의 Alcalase®2.4L와 FlavourzymeTM 500 MG, ProtametTM, Neutrase®로, 사용된 효소의 특징은 하기 표 2와 같다. 효소별 단백질 함량 측정은 대조군(효소 무처리) Alcalase®2.4L와 FlavourzymeTM 500 MG, ProtametTM를 이용하였다. 귀뚜라미 효소가수분해물은 원료 귀뚜라미 착유박과 물을 1:10(w/w)의 비율로 혼합하여 3종의 상업적 효소와 혼합 효소를 원료 대비 2% (w/w) 첨가하고 폴리에틸렌 필름 포장지에 담아 밀봉하였으며, 밀봉된 효소용액을 고압액화기 (TFS-20, Innoway, Incheon, Korea)에 넣고 50℃로 온도를 조절한 항온수조 (Wisd WB-11, Daiham, Seoul, Korea)에 동결보존용 튜브에 밀봉된 효소용액을 넣고 4시간 동안 반응시켰다. 그 결과를 하기 표 3에 나타내었다.Commercial enzymes used in the present invention are Novozymes (Novozymes, Bagsvard, Denmark) of Alcalase® 2.4L and Flavorzyme TM 500 MG, Protamet TM , Neutrase®, and the characteristics of the enzymes used are shown in Table 2 below. For the measurement of protein content by enzyme, control (no enzyme treatment) Alcalase® 2.4L, Flavorzyme TM 500 MG, and Protamet TM were used. For the cricket enzyme hydrolyzate, raw cricket milk meal and water are mixed in a ratio of 1:10 (w/w), 3 commercial enzymes and 2% (w/w) of the mixed enzyme are added compared to the raw material and put in polyethylene film wrapper. The sealed, sealed enzyme solution was placed in a high-pressure liquefier (TFS-20, Innoway, Incheon, Korea) and a tube for cryopreservation in a constant temperature water tank (Wisd WB-11, Daiham, Seoul, Korea) controlled to 50 °C. The sealed enzyme solution was added and reacted for 4 hours. The results are shown in Table 3 below.

이름name 기원origin 최적 pHoptimum pH 적정온도(℃)Appropriate temperature (℃) 활성단위active unit 반응기작reaction mechanism Alcalase®2.4 LAlcalase® 2.4 L B. licheniformisB. licheniformis 6.5~8.56.5~8.5 55~7055-70 2.4 AU/g2.4 AU/g endoendo FlavourzymeFlavorzyme TMTM 500 MG 500 mg A. oryzaeA. oryzae 5.0~7.05.0~7.0 5050 500 LAPU/g500 LAPU/g endo, exoendo, exo Neutrase®Neutrase® Bacillus amyloliiensquenfaciensBacillus amyloliiensquenfaciens 5.5~7.05.5~7.0 35~6035 to 60 1.5 AU1.5 AU endoendo ProtamexProtamex TMTM BacillusBacillus 5.5~7.55.5~7.5 35~6035 to 60 1.5 AU1.5 AU endoendo

AU: Anson Unit, LAPU: leucine aminopeptidase unitAU: Anson Unit, LAPU: leucine aminopeptidase unit

0.1 MPa0.1 MPa 50 MPa50 MPa 압력에 대한 차이difference in pressure 대조군control 0.680.68 0.710.71 0.030.03 Alcalase®2.4 LAlcalase® 2.4 L 4.494.49 6.736.73 2.242.24 FlavourzymeFlavorzyme TMTM 500 MG 500 mg 4.564.56 6.226.22 1.661.66 ProtamexProtamex TMTM 5.945.94 7.247.24 1.301.30

(단위: %)(unit: %)

쌍별귀뚜라미의 효율적인 단백질 가수분해물 제조를 위해 효소별 단백질 함량을 분석한 결과, 건조 쌍별귀뚜라미에 대한 효소처리액의 단백질 함량은 무처리에서 0.1MPa, 50MPa 각각 0.68, 0.71로 나타났다. 쌍별귀뚜라미에 대한 효소별 단백질 함량을 분석한 결과 0.1MPa과 50MPa에서 Protamex가 모두 가장 높은 5.94%, 7.24%의 함량을 나타냈다. 그러나 고압에 대한 효율성 증가는 Alcalase가 0.1MPa에서 4.49% 였으나, 50MPa에서는 6.73으로 고압으로 될수록 높아지는 것을 확인하였다.As a result of analyzing the protein content for each enzyme for the efficient production of protein hydrolysates of crickets, the protein content of the enzyme-treated solution for dry crickets was 0.68 and 0.71, respectively, at 0.1 MPa and 50 MPa in the untreated case. As a result of analyzing the protein content for each enzyme in paired crickets, Protamex showed the highest content of 5.94% and 7.24% at 0.1 MPa and 50 MPa. However, the efficiency increase for high pressure was 4.49% at 0.1 MPa for Alcalase, but 6.73 at 50 MPa, confirming that it increased as the pressure increased.

<실시예 3> 효소처리 농도에 따른 쌍별귀뚜라미의 단백질 가수분해<Example 3> Protein hydrolysis of paired crickets according to the concentration of enzyme treatment

상기 실시예 1-2에서 제조한 탈지 쌍별귀뚜라미박 200 g에 증류수를 10배 가수하여 4종의 단백질 분해효소를 순서에 상관 없이 동시에 각각 0, 0.10, 0.15 및 0.20%(w/w) 첨가한 후 55~60℃에서 100 rpm, 60분 교반 및 반응시켰다.Distilled water was added 10 times to 200 g of the defatted pairwise cricket meal prepared in Example 1-2, and 0, 0.10, 0.15 and 0.20% (w/w) were added at the same time, regardless of the order, of 4 proteolytic enzymes. Then, it was stirred and reacted at 100 rpm, 60 minutes at 55 ~ 60 ℃.

가수분해 종료 후 95℃에서 10분간 불활성한 후 원심분리(4,000 rpm, 10분)하여 상등액을 얻었다. 상등액의 °Brix, 가용성 고형분 함량, 조단백 함량, 아미노태질소 함량, 탁도, 갈색도를 분석하여 하기 표 4에 나타내었다. 결과값은 3번 실험한 결과의 평균값으로 나타내었다. 각각의 분석 방법은 아래와 같다.After the hydrolysis was completed, it was inactivated at 95° C. for 10 minutes, and then centrifuged (4,000 rpm, 10 minutes) to obtain a supernatant. °Brix, soluble solids content, crude protein content, amino nitrogen content, turbidity, and brownness of the supernatant were analyzed and shown in Table 4 below. The result value is shown as the average value of the results of three experiments. Each analysis method is as follows.

실시예 3-1: 효소 0% 첨가Example 3-1: Addition of enzyme 0%

실시예 3-2: 효소 0.10% 첨가Example 3-2: 0.10% enzyme addition

실시예 3-3: 효소 0.15% 첨가Example 3-3: 0.15% enzyme addition

실시예 3-4: 효소 0.20% 첨가Example 3-4: 0.20% enzyme addition

3.1. 가용성 및 수용성 고형분 함량 분석3.1. Analysis of soluble and water-soluble solids content

가용성 고형분 함량은 1 mL를 취하여 당도계를 이용하여 측정하였으며, °Brix로 표시하였으며, 수용성 고형분 함량은 상압가열건조법으로 수분함량을 측정한 후 100%에서 수분함량을 뺀 고형분 함량으로 조사하였다.The content of soluble solids was measured using a saccharometer by taking 1 mL, and expressed as °Brix. The content of soluble solids was investigated as the solid content by subtracting the moisture content from 100% after measuring the moisture content by atmospheric heating and drying method.

3.2. 조단백 함량 분석3.2. Analysis of crude protein content

조단백 함량은 식품공전의 일반시험법(조단백질)을 참고하여 자동단백질분석기(FoodALTY)를 사용하여 측정하였다. 검체 1 g을 정밀하게 취하여 분해튜브에 넣고 분해촉진제 0.5g과 진한 황산 12 mL을 넣은 후 420℃의 분해장치에서 45~60분간 분해하여 분해액의 색이 투명한 연푸른 또는 투명한 노란색이 되면 상온으로 냉각시켰다. 분해된 시험용액에 80 mL의 증류수를 주의하여 첨가한 후 증류장치에 넣은 후 25 mL의 혼합지시약(1% 붕산용액), 40% 수산화나트륨용액 50 mL을 첨가하여 자동장치에 의해 증류(3~4min)한 후 0.1N 염산용액으로 적정하였다.Crude protein content was measured using an automatic protein analyzer (FoodALTY) referring to the general test method (crude protein) of the Food Code. Take 1 g of the sample precisely, put it in a decomposition tube, add 0.5 g of decomposition accelerator and 12 mL of concentrated sulfuric acid, and decompose in a decomposition device at 420 ° C for 45 to 60 minutes. cooled. After carefully adding 80 mL of distilled water to the decomposed test solution, put it in a distillation apparatus, add 25 mL of mixing indicator (1% boric acid solution), and 50 mL of 40% sodium hydroxide solution, and distill by an automatic device (3~ 4 min) and titrated with 0.1N hydrochloric acid solution.

조단백질(%)=(소비된 HCl 양- 공시험 HCL)/검체량(mg) x M x 14.01/검체량(mg) x F x 100Crude protein (%) = (amount of HCl consumed - blank test HCL) / sample amount (mg) x M x 14.01 / sample amount (mg) x F x 100

M: HCl 몰 농도M: HCl molarity

14.01: 질소의 원자량14.01: atomic weight of nitrogen

F: 질소계수 (6.25)F: nitrogen coefficient (6.25)

3.3. 아미노태 질소 함량 분석3.3. Analysis of amino nitrogen content

아미노태 질소 측정은 Formol 법을 이용하여 검체 0.5 g을 취해 증류수를 이용하여 50 mL을 정용하고 1분간 균질화한 후 원심분리(4,000 rpm, 10min)한 후 상등액을 시료로 하였다. 상등액 5 mL에 중성 formalin 용액 10 mL을 가하고 0.1 N NaOH 용액으로 pH 8.3이 되도록 중화적정하였다. 공시험은 상등액 5 mL 대신 증류수를 넣어 시료에서와의 같은 방법으로 실험을 시행하여 아미노태 질소 함량을 구하였다.For amino nitrogen measurement, 0.5 g of a sample was taken using the Formol method, 50 mL of distilled water was used, homogenized for 1 minute, centrifuged (4,000 rpm, 10 min), and the supernatant was used as a sample. 10 mL of neutral formalin solution was added to 5 mL of the supernatant, and neutralized titration was performed with 0.1 N NaOH solution to pH 8.3. In the blank test, distilled water was added instead of 5 mL of the supernatant, and the experiment was conducted in the same manner as in the sample to determine the amino acid nitrogen content.

아미노태질소(%)=(소비된 NaOH 양- 공시험 NaOH) x F x 1.4 x D/시료의 채취량(g) x 100Amino nitrogen (%) = (NaOH consumed- NaOH blank test) x F x 1.4 x D/Amount of sample collected (g) x 100

F: 0.1 N NaOH 용액의 역가F: titer of 0.1 N NaOH solution

1.4: 0.1 N NaOH 용액 1 mL에 상당하는 질소량1.4: Amount of nitrogen equivalent to 1 mL of 0.1 N NaOH solution

D: 희석배수D: dilution factor

3.4. 탁도 및 갈색도3.4. Turbidity and Brownness

탁도는 whatman filter paper로 여과한 단백질 가수분해물을 시료로 660 nm에서 흡광도를 측정하였으며, 갈색도는 동일한 방법으로 여과한 단백질 가수분해물을 시료로 453 nm에서 흡광도를 측정하였다.For turbidity, absorbance was measured at 660 nm using a protein hydrolyzate filtered with whatman filter paper, and absorbance was measured at 453 nm for a protein hydrolyzate filtered by the same method for brownness.

실시예 3-1Example 3-1 실시예 3-2Example 3-2 실시예 3-3Example 3-3 실시예 3-4Example 3-4 °Brix°Brix 2.502.50 5.435.43 5.905.90 6.176.17 가용성 고형분(%)Soluble solids (%) 2.102.10 4.644.64 5.055.05 5.795.79 조단백(%)Crude protein (%) 1.031.03 3.023.02 3.133.13 3.393.39 아미노태질소(%)Amino nitrogen (%) 0.070.07 0.100.10 0.140.14 0.150.15 탁도turbidity 2.302.30 4.234.23 3.103.10 3.003.00 갈색도brown degree 3.373.37 6.776.77 5.205.20 4.934.93

효소 처리 농도 (0, 0.10, 0.15 및 0.20%(w/w))에 따른 쌍별귀뚜라미 단백질 가수분해물의 품질특성을 분석을 분석한 결과, 쌍별귀뚜라미 단백질 가수분해추출물의 가용성 고형분 함량은 2.50~6.17 °Brix로, 첨가되는 효소의 농도가 높아질수록 증가하였으며, 수용성 고형분 또한 2.10~5.79%로, 농도가 높아질수록 증가하는 경향을 나타내었다. 조단백 함량은 1.03~3.39%로 첨가되는 효소의 농도가 높아질수록 증가하였으며, 아미노태 질소?t량 또한 0.07~0.15%로 농도가 높아질수록 증가하는 경향을 나타내었다. 효소처리 농도에 따른 쌍별귀뚜라미 단백질 가수분해물의 탁도는 실시예 3-2 (0.1%첨가)구간에서 4.23으로 높은 값을 나타내었으며, 그 다음 실시예 3-3 (0.15%첨가) 3.10, 실시예 3-4 (0.20%첨가) 3.00 순으로 나타났으며, 갈색도 또한 실시예 3-2 구간이 6.77로 가장 높은 값을 나타내었으며, 그 다음 실시예 3-4 5.20, 실시예 3-5 4.93 순으로 나타났다. 따라서 쌍별귀뚜라미 단백질 가수분해물 제조시 첨가되는 효소의 농도가 높아짐에 따라 고형분(가용성 및 수용성) 및 조단백, 아미노태 질소 함량이 높아지는 경향으로 나타나 단백질 추출수율을 높이기 위해서는 효소처리 농도를 0.20%(w/w)로 설정하는 것이 적합한 것으로 나타났다.As a result of analyzing the quality characteristics of the hydrolyzed cricket protein according to the enzyme treatment concentration (0, 0.10, 0.15 and 0.20% (w/w)), the soluble solids content of the hydrolyzed cricket protein extract was 2.50~6.17 ° As Brix, it increased as the concentration of the added enzyme increased, and the water-soluble solid content was also 2.10 to 5.79%, showing a tendency to increase as the concentration increased. The crude protein content was 1.03~3.39%, and it increased as the concentration of the added enzyme increased, and the amount of amino nitrogen?t was also 0.07~0.15%, showing a tendency to increase as the concentration increased. The turbidity of the cricket protein hydrolyzate according to the concentration of enzyme treatment showed a high value of 4.23 in the section of Example 3-2 (0.1% added), followed by Example 3-3 (0.15% added) 3.10, Example 3 -4 (0.20% added) appeared in the order of 3.00, and the brown color also showed the highest value at 6.77 in the section of Example 3-2, followed by Examples 3-4 5.20 and Examples 3-5 4.93 appear. Therefore, as the concentration of the enzyme added during the production of the protein hydrolyzate of paired cricket increases, the solid content (soluble and water-soluble), crude protein, and amino nitrogen content tend to increase. w) was found to be suitable.

<실시예 4> 효소처리 시간에 따른 쌍별귀뚜라미의 단백질 가수분해<Example 4> Protein hydrolysis of paired crickets according to enzyme treatment time

상기 실시예 1-2에서 제조한 탈지 쌍별귀뚜라미박 200 g에 증류수를 10배 가수하여 4종의 단백질 분해효소를 순서에 상관없이 동시에 0.20w/w%를 첨가한 후 55~60℃에서 100 rpm으로 30, 60 및 90분 각각 교반 및 반응시켰다. 가수분해 종료 후 95℃에서 10분간 불활성한 후 원심분리(4,000 rpm, 10분)하여 상등액을 얻었다. 상기 실시예 3과 마찬가지로, 상등액의 °Brix, 가용성 고형분 함량, 조단백 함량, 아미노태질소 함량, 탁도, 갈색도를 분석하여 하기 표 5에 나타내었다. 결과값은 3번 실험한 결과의 평균값으로 나타내었다.Distilled water was added 10 times to 200 g of the defatted twin-star cricket meal prepared in Example 1-2, and 0.20 w/w% of 4 proteolytic enzymes were simultaneously added in any order, and then at 55-60 ° C. at 100 rpm. was stirred and reacted for 30, 60 and 90 minutes, respectively. After the hydrolysis was completed, it was inactivated at 95° C. for 10 minutes, and then centrifuged (4,000 rpm, 10 minutes) to obtain a supernatant. As in Example 3, °Brix, soluble solids content, crude protein content, amino nitrogen content, turbidity, and brownness of the supernatant were analyzed and shown in Table 5 below. The result value is shown as the average value of the results of three experiments.

실시예 4-1: 효소처리 30분Example 4-1: Enzyme treatment 30 minutes

실시예 4-2: 효소처리 60분Example 4-2: Enzyme treatment 60 minutes

실시예 4-3: 효소처리 90분Example 4-3: Enzyme treatment for 90 minutes

실시예 4-1Example 4-1 실시예 4-2Example 4-2 실시예 4-3Example 4-3 °Brix°Brix 5.035.03 6.176.17 7.177.17 가용성 고형분(%)Soluble solids (%) 3.963.96 5.795.79 5.745.74 조단백(%)Crude protein (%) 2.752.75 3.393.39 3.943.94 아미노태질소(%)Amino nitrogen (%) 0.120.12 0.150.15 0.180.18 탁도turbidity 1.901.90 3.003.00 2.632.63 갈색도brown degree 3.403.40 4.934.93 4.404.40

효소처리 시간(30, 60 및 90분)에 따른 쌍별귀뚜라미 단백질 가수분해물의 품질특성을 분석한 결과, 쌍별귀뚜라미 단백질 가수분해물의 가용성 고형분 함량은 5.03~7.17 °Brix로 효소반응 시간이 길어질수록 증가하였으며, 수용성 고형분 또한 3.96~5.79%로 시간이 길어질수록 증가하는 경향을 보였다. 조단백 함량은 2.75~3.94%로 시간이 경과함에 따라 증가하는 경향을 나타내었으며, 아미노태질소 함량 또한 0.12~0.18%로 나타났다. 효소처리 시간에 따른 쌍별귀뚜라미 단백질 가수분해물의 탁도는 실시예 4-2 (60분처리) 구간이 3.00으로 높은 값을 나타내었으며, 그 다음 실시예 4-3 (90분처리), 실시예 4-1 (30분처리)순으로 나타났으며, 갈색도 또한 실시예 4-2 구간이 4.93, 실시예 4-3 4.40, 실시예 4-1 3.40 순으로 나타났다. 따라서 쌍별귀뚜라미 단백질 가수분해물 제조시 효소반응 시간이 증가함에 따라 고형분(가용성 및 수용성) 및 조단백, 아미노태질소 함량이 증가하는 경향을 나타내었으나, 산업에서의 시간적 및 비용적 범위를 고려하여 효소반응 시간을 60분 처리하는 것으로 하였다.As a result of analyzing the quality characteristics of the hydrolyzate of cricket protein according to the enzyme treatment time (30, 60 and 90 minutes), the soluble solids content of the hydrolyzate of cricket protein was 5.03~7.17 °Brix, which increased as the enzyme reaction time increased. , and the water-soluble solid content was also 3.96-5.79%, showing a tendency to increase as the time increased. The crude protein content was 2.75~3.94%, and it showed a tendency to increase with the lapse of time, and the amino nitrogen content was also 0.12~0.18%. The turbidity of the cricket protein hydrolyzate according to the enzyme treatment time showed a high value of 3.00 in the section of Example 4-2 (60 minutes treatment), followed by Examples 4-3 (90 minutes treatment), Example 4- 1 (30-minute treatment) was shown, and brown was also shown in the order of Example 4-2 section 4.93, Example 4-3 4.40, and Example 4-1 3.40. Therefore, as the enzymatic reaction time increased, the solids (soluble and water-soluble), crude protein, and amino nitrogen content tended to increase when preparing the hydrolyzate of cricket protein. was treated for 60 minutes.

<실시예 5> 효소처리 순서에 따른 쌍별귀뚜라미의 단백질 가수분해<Example 5> Protein hydrolysis of paired crickets according to the enzymatic treatment sequence

탈지 쌍별귀뚜라미박 200 g에 증류수를 10배 가수하여 4종의 단백질 분해효소 각각 0.20%를 첨가후 55~60℃에서 100 rpm으로 60분 동안 교반 및 반응시켰다. 이때, 효소는 4개의 효소를 factorial design으로 효소처리 순서를 달리하여 총 24개의 처리구로 제조하였음. 가수분해 종료 후 95℃에서 10분간 불활성한 후 원심분리(4,000 rpm, 10분)하여 상등액을 얻었다. 상기 실시예 3과 같은 방법으로 분석한 효소처리 순서에 따른 단백질 가수분해물의 pH, Brix, 가용성 고형분 함량을 하기 표 6에 나타내고, 조단백 함량, 아미노태질소 함량은 표 7, 탁도 및 갈색도는 표 8에 나타내었다. Distilled water was added 10 times to 200 g of defatted twin-star cricket meal, 0.20% of each of the four proteolytic enzymes was added, and then stirred and reacted at 55 to 60° C. at 100 rpm for 60 minutes. At this time, 4 enzymes were prepared in a total of 24 treatment groups by changing the order of enzyme treatment by factorial design. After the hydrolysis was completed, it was inactivated at 95° C. for 10 minutes, and then centrifuged (4,000 rpm, 10 minutes) to obtain a supernatant. The pH, Brix, and soluble solids content of the protein hydrolyzate according to the enzymatic treatment sequence analyzed in the same manner as in Example 3 are shown in Table 6 below, the crude protein content and amino nitrogen content are Table 7, and the turbidity and brownness are Tables 8 is shown.

실시예 5-1 내지 5-24: 4개의 효소 Alcalase®(A), FlavourzymeTM(F), Neutrase®(N), ProtamexTM(P)를 이용하여 24개의 효소 처리 순서를 조합하였다. 예를들어, FNAP는 FlavourzymeTM, Neutrase®, Alcalase®, ProtamexTM 순으로 처리한 것이다.Examples 5-1 to 5-24: 24 enzyme treatment sequences were combined using 4 enzymes Alcalase® (A), Flavourzyme ™ (F), Neutrase® (N), Protamex ™ (P). For example, FNAP is treated with Flavorzyme TM , Neutrase ® , Alcalase ® , and Protamex TM in that order.

효소처리순서Enzyme treatment sequence pHpH BrixBrix 가용성 고형분(%)Soluble solids (%) 실시예 5-1Example 5-1 FNAPFNAP 6.276.27 5.235.23 6.586.58 실시예 5-2Example 5-2 FNPAFNPA 6.236.23 6.376.37 6.336.33 실시예 5-3Example 5-3 FANPFANP 6.286.28 6.236.23 6.796.79 실시예 5-4Example 5-4 FAPNFAPN 6.256.25 6.276.27 6.956.95 실시예 5-5Example 5-5 FPNAFPNA 6.246.24 4.674.67 4.744.74 실시예 5-6Example 5-6 FPANFPAN 6.266.26 6.436.43 6.536.53 실시예 5-7Example 5-7 NFAPNFAP 6.246.24 5.305.30 5.455.45 실시예 5-8Examples 5-8 NFPANFPA 6.266.26 4.604.60 4.334.33 실시예 5-9Examples 5-9 NAFPNAFP 6.246.24 5.835.83 5.635.63 실시예 5-10Examples 5-10 NAPFNAPF 6.236.23 6.736.73 6.366.36 실시예 5-11Examples 5-11 NPFANPFA 6.236.23 5.705.70 5.015.01 실시예 5-12Examples 5-12 NPAFNPAF 6.236.23 5.535.53 5.455.45 실시예 5-13Examples 5-13 AFNPAFNP 6.226.22 6.836.83 7.137.13 실시예 5-14Examples 5-14 AFPNAFPN 6.206.20 5.835.83 6.676.67 실시예 5-15Examples 5-15 ANFPANFP 6.216.21 6.136.13 6.146.14 실시예 5-16Examples 5-16 ANPFANPF 6.176.17 6.076.07 5.745.74 실시예 5-17Examples 5-17 APFNAPFN 6.216.21 6.306.30 6.826.82 실시예 5-18Examples 5-18 APNFAPNF 6.216.21 6.336.33 6.816.81 실시예 5-19Examples 5-19 PFNAPFNA 6.176.17 5.605.60 5.605.60 실시예 5-20Examples 5-20 PFANPFAN 6.206.20 4.934.93 5.265.26 실시예 5-21Example 5-21 PNFAPNFA 6.196.19 5.875.87 5.615.61 실시예 5-22Example 5-22 PNAFPNAF 6.196.19 5.305.30 5.595.59 실시예 5-23Example 5-23 PAFNPAFN 6.186.18 5.435.43 5.685.68 실시예 5-24Example 5-24 PANFPANF 6.196.19 5.675.67 5.945.94

효소처리 순서에 따른 쌍별귀뚜라미 단백질 가수분해물의 품질특성을 분석한 결과, 24개의 쌍별귀뚜라미 단백질 가수분해추출물의 pH는 6.17~6.28 범위로 중성을 나타내었다. 가용성 고형분 함량은 4.60~6.83 °Brix로 NFPA 처리구에서 4.60 °Brix로 가장 낮은 값을, ANFP 처리구에서 6.83 °Brix로 가장 높은 값을 나타내었으며, 수용성 고형분 함량은 4.74~7.13%로 나타났다. 이상의 결과 가용성 고형분 함량 및 고형분 함량에서 효소의 처리 순서가 큰 영향을 미치지 않는 것으로 나타났다.As a result of analyzing the quality characteristics of the hydrolyzed cricket protein according to the enzymatic treatment sequence, the pH of 24 cricket protein hydrolyzate extracts was neutral in the range of 6.17 to 6.28. Soluble solids content was 4.60~6.83 °Brix, the lowest value at 4.60 °Brix in the NFPA treatment group, and the highest value at 6.83 °Brix in the ANFP treatment group, and the soluble solid content was 4.74~7.13%. As a result, it was found that the soluble solid content and the order of the enzyme treatment did not have a significant effect on the solid content.

효소처리순서Enzyme treatment sequence 조단백(%)Crude protein (%) 아미노태질소(%)Amino nitrogen (%) 실시예 5-1Example 5-1 FNAPFNAP 4.174.17 0.080.08 실시예 5-2Example 5-2 FNPAFNPA 4.044.04 0.040.04 실시예 5-3Example 5-3 FANPFANP 4.414.41 0.040.04 실시예 5-4Example 5-4 FAPNFAPN 3.893.89 0.050.05 실시예 5-5Example 5-5 FPNAFPNA 2.782.78 0.070.07 실시예 5-6Example 5-6 FPANFPAN 4.634.63 0.070.07 실시예 5-7Example 5-7 NFAPNFAP 3.173.17 0.080.08 실시예 5-8Examples 5-8 NFPANFPA 2.482.48 0.080.08 실시예 5-9Examples 5-9 NAFPNAFP 4.284.28 0.070.07 실시예 5-10Examples 5-10 NAPFNAPF 4.464.46 0.080.08 실시예 5-11Examples 5-11 NPFANPFA 3.263.26 0.100.10 실시예 5-12Examples 5-12 NPAFNPAF 3.463.46 0.100.10 실시예 5-13Examples 5-13 AFNPAFNP 4.264.26 0.060.06 실시예 5-14Examples 5-14 AFPNAFPN 3.743.74 0.070.07 실시예 5-15Examples 5-15 ANFPANFP 3.633.63 0.060.06 실시예 5-16Examples 5-16 ANPFANPF 4.044.04 0.060.06 실시예 5-17Examples 5-17 APFNAPFN 3.843.84 0.080.08 실시예 5-18Examples 5-18 APNFAPNF 4.434.43 0.080.08 실시예 5-19Examples 5-19 PFNAPFNA 3.763.76 0.110.11 실시예 5-20Examples 5-20 PFANPFAN 3.333.33 0.080.08 실시예 5-21Example 5-21 PNFAPNFA 4.154.15 0.080.08 실시예 5-22Example 5-22 PNAFPNAF 2.432.43 0.140.14 실시예 5-23Example 5-23 PAFNPAFN 3.563.56 0.130.13 실시예 5-24Example 5-24 PANFPANF 2.312.31 0.140.14

24개의 쌍별귀뚜라미 단백질 가수분해추출물의 조단백 함량은 2.31~4.63%의 범위로 나타났으며, 그 중 실시예 5-6 (FPAN) 처리구가 4.63%로 가장 높은 값을 나타내었다. 아미노태 질소 함량의 경우 0.04~0.14%의 범위로 실시예 5-22 (PNAF) 및 실시예 5-24 (PANF) 처리구가 0.14%로 가장 높게 나타났다. The crude protein content of the 24 cricket protein hydrolyzate extracts was found to be in the range of 2.31 to 4.63%, and among them, Example 5-6 (FPAN) treatment showed the highest value at 4.63%. In the case of amino nitrogen content, in the range of 0.04 to 0.14%, Example 5-22 (PNAF) and Example 5-24 (PANF) treated groups were the highest at 0.14%.

효소처리순서Enzyme treatment sequence 탁도turbidity 갈색도brown degree 실시예 5-1Example 5-1 FNAPFNAP 0.170.17 1.351.35 실시예 5-2Example 5-2 FNPAFNPA 0.170.17 1.611.61 실시예 5-3Example 5-3 FANPFANP 0.220.22 1.691.69 실시예 5-4Example 5-4 FAPNFAPN 0.220.22 1.581.58 실시예 5-5Example 5-5 FPNAFPNA 0.180.18 1.491.49 실시예 5-6Example 5-6 FPANFPAN 0.300.30 1.901.90 실시예 5-7Example 5-7 NFAPNFAP 0.240.24 1.401.40 실시예 5-8Examples 5-8 NFPANFPA 0.150.15 1.081.08 실시예 5-9Examples 5-9 NAFPNAFP 0.270.27 1.831.83 실시예 5-10Examples 5-10 NAPFNAPF 0.280.28 2.032.03 실시예 5-11Examples 5-11 NPFANPFA 0.270.27 1.631.63 실시예 5-12Examples 5-12 NPAFNPAF 0.270.27 1.681.68 실시예 5-13Examples 5-13 AFNPAFNP 0.410.41 1.971.97 실시예 5-14Examples 5-14 AFPNAFPN 0.330.33 1.841.84 실시예 5-15Examples 5-15 ANFPANFP 0.330.33 1.921.92 실시예 5-16Examples 5-16 ANPFANPF 0.170.17 1.421.42 실시예 5-17Examples 5-17 APFNAPFN 0.390.39 2.082.08 실시예 5-18Examples 5-18 APNFAPNF 0.340.34 2.062.06 실시예 5-19Examples 5-19 PFNAPFNA 0.300.30 1.661.66 실시예 5-20Examples 5-20 PFANPFAN 0.210.21 1.451.45 실시예 5-21Example 5-21 PNFAPNFA 0.190.19 1.621.62 실시예 5-22Example 5-22 PNAFPNAF 0.130.13 1.131.13 실시예 5-23Example 5-23 PAFNPAFN 0.160.16 1.371.37 실시예 5-24Example 5-24 PANFPANF 0.190.19 1.481.48

24개의 쌍별귀뚜라미 단백질 가수분해추출물의 탁도는 0.13~0.39로 나타났으며, 갈색도의 경우 1.13~2.08로 나타났다. 탁도와 갈색도는 효소 처리의 순서에 의한 영향이 미미한 것으로 확인하였다.The turbidity of the protein hydrolyzed extracts of 24 pairs of crickets was 0.13 to 0.39, and the brownness was 1.13 to 2.08. It was confirmed that the turbidity and brownness were insignificantly affected by the order of the enzyme treatment.

상기 실시예 3 내지 실시예 5의 결과를 종합한 결과, 쌍별귀뚜라미 단백질 가수분해 제조시 단백질 분해효소는 Flavourzyme, Protamex, Alcalase 및 Neutrase순으로 각각 0.2%(w/w) 첨가하여 55~60℃에서 60분간 반응시키는 것이 가장 적합한 것으로 확인하였다(도 1).As a result of synthesizing the results of Examples 3 to 5, the proteolytic enzymes were added in the order of Flavorzyme, Protamex, Alcalase, and Neutrase in the order of 0.2% (w/w), respectively, at 55 to 60 ° C. It was confirmed that the reaction for 60 minutes was most suitable (FIG. 1).

<실시예 6> 쌍별귀뚜라미 단백농축액의 제조<Example 6> Preparation of Ssangbyeol Cricket Protein Concentrate

상기 실시예 5에서 제조한 쌍별귀뚜라미 단백질 추출물을 농축하기 위하여 회전식 감압 농축기에서 온도 60℃, 회전속도 40 rpm 조건 농축을 실시하였으며, 농축 종말점은 농축액의 농도가 40°Brix를 나타낼 때로 하였다.In order to concentrate the bi-star cricket protein extract prepared in Example 5, concentration was carried out in a rotary vacuum concentrator at a temperature of 60° C. and a rotation speed of 40 rpm, and the concentration end point was determined when the concentration of the concentrate was 40° Brix.

<실시예 7> 쌍별귀뚜라미 단백농축분말의 제조<Example 7> Preparation of double star cricket protein concentrate powder

상기 실시예 6에서 제조한 쌍별귀뚜라미 단백농축액을 건조하여 단백농축분말을 제조하기 위하여, 분무건조기를 사용하였으며, 분무건조 조건은 In 200℃, Out 100℃ 조건으로 분무건조를 실시하였다.In order to prepare a protein concentrate powder by drying the double star cricket protein concentrate prepared in Example 6, a spray dryer was used, and spray drying was carried out under the conditions of In 200 ℃ and Out 100 ℃.

<실험예> 쌍별귀뚜라미 원료 및 탈지박을 이용한 단백추출물의 대사체 분석<Experimental Example> Metabolite analysis of protein extracts using raw materials for twin-star crickets and skim foil

분석 대상 시료는 쌍별 귀뚜라미 단백농축액 제조공정에 따른 4가지 시료로, 1) 쌍별귀뚜라미 건조원료, 2) 쌍별귀뚜라미 착유박(실시예 1-2), 3) 단백가수분해 효소처리한 쌍별귀뚜라미 단백농축액(실시예 6), 4) 쌍별귀뚜라미 단백농축분말(실시예 7)이며, 추출은 70% 메탄올로 정량을 위한 ITSD로 zidovudine, 0.02 g/mL을 사용하였다.The samples to be analyzed are four samples according to the production process of the pairwise cricket protein concentrate, 1) the pairwise cricket dry raw material, 2) the pairwise cricket milk meal (Example 1-2), 3) the pairwise cricket protein concentrate treated with proteolytic enzyme (Example 6), 4) Concentrated cricket protein powder (Example 7), extraction was 70% methanol, and zidovudine, 0.02 g/mL was used as ITSD for quantification.

분석장비는 UPLC-Q-TOF MS (Vion, Waters, Milford, MA, USA), LC 조건은 inject volume : 1 uL; Column : Acquity UPLC BEH C18 column (2.1 mm ×100 mm, 1.7 im; Waters); Solvent A: water with 0.1% FA; Solvent B: ACN with 0.1% FA; Flow rate: 0.35 mL/min, MS 조건은, ESI-negative mode; Capillary voltage : 2.5 kV; Sample cone voltage : 20 V; Desolvation gas flow : 900 L/h; Cone gas flow : 30 L/h; Desolvation temperature : 400 ℃; Ion source temperatgure : 100 ℃; Data collection : m/z 50-1500 range with a scan time of 0.5 s; Lock mass : Leucine-enkephalin (554.2615Da); Quality control (QC), a mixture of all samples was injected after every 10 samples이다. 분석결과 데이터 처리는 UNIFI version 1.8.2.169 (Waters)을 통해 Data collection, normalization, alignment 과정을 거치고 물질동정은 Chemspider database, Metlin database (metlin.scripps.edu) and human metabolome databases (www.hmdb.ca), Ezmass database, authentic standards를 통해 이루어졌다.Analysis equipment is UPLC-Q-TOF MS (Vion, Waters, Milford, MA, USA), LC conditions are inject volume: 1 uL; Column: Acquity UPLC BEH C18 column (2.1 mm × 100 mm, 1.7 im; Waters); Solvent A: water with 0.1% FA; Solvent B: ACN with 0.1% FA; Flow rate: 0.35 mL/min, MS conditions, ESI-negative mode; Capillary voltage: 2.5 kV; Sample cone voltage: 20 V; Desolvation gas flow: 900 L/h; Cone gas flow: 30 L/h; Desolvation temperature: 400℃; Ion source temperature: 100℃; Data collection: m/z 50-1500 range with a scan time of 0.5 s; Lock mass: Leucine-enkephalin (554.2615 Da); Quality control (QC), a mixture of all samples was injected after every 10 samples. Data processing of analysis results is data collection, normalization, and alignment process through UNIFI version 1.8.2.169 (Waters), and substance identification is conducted in Chemspider database, Metlin database (metlin.scripps.edu) and human metabolome databases (www.hmdb.ca) , Ezmass database, and authentic standards.

쌍별귀뚜라미 건조원료, 본 발명에서 제공하는 착유 공정에 따라 제조한 쌍별귀뚜라미 착유박, 본 발명에서 제공하는 단백가수분해 효소처리 한 쌍별귀뚜라미 단백농축액, 단백농축분말이 대사체를 분석한 결과를 하기 표 9 내지 표 11 및 도 2에 나타내고, 크로마토그래피로 분석한 크로마토그램을 도 3에 나타내었다.The table below shows the results of metabolite analysis of the dried raw material of twin-star crickets, the milk-milking meal of twin-star crickets prepared according to the milking process provided by the present invention, the protein-hydrolytic enzyme-treated twin-star cricket protein concentrate and the protein concentrate powder provided in the present invention. 9 to Table 11 and FIG. 2, and chromatograms analyzed by chromatography are shown in FIG.

쌍별귀뚜라미 제조공정 변화에 따른 대사체 성분 상대량 변화 (원물→착유박)Changes in the relative amount of metabolite components according to the change in the manufacturing process of twin-star crickets (raw material → milk meal) CompoundsCompounds RTRT 착유공정 후 After milking process
상대 증감율(%)Relative increase/decrease rate (%) 증가increase xanthinexanthine 1.25611.2561 1.5952 1.5952 tyrosinetyrosine 1.4471.447 1.5554 1.5554 phenylalaninephenylalanine 2.50142.5014 0.6590 0.6590 9,12,13-TriHOME9,12,13-TriHOME 4.25054.2505 6.4402 6.4402 13-Hydroxyoctadecadienoic acid13-Hydroxyoctadecadienoic acid 5.76465.7646 18.2229 18.2229 hydroxystearic acidhydroxystearic acid 6.26086.2608 10.6904 10.6904 감소decrease uric aciduric acid 1.09891.0989 -0.6694 -0.6694 acetyl-glutamic acidacetyl-glutamic acid 1.49151.4915 -0.1761 -0.1761 dopamine sulfatedopamine sulfate 2.05872.0587 -0.1103 -0.1103 arabinosylhypoxanthinearabinosylhypoxanthine 2.32592.3259 -2.7505 -2.7505 sulfotyrosinesulfotyrosine 2.74052.7405 -2.8563 -2.8563 methyl sulfotyrosinatemethyl sulfotyrosinate 2.80072.8007 -0.1286 -0.1286 Di(buta-2,3-dienyl)butane-1,4-diamineDi(buta-2,3-dienyl)butane-1,4-diamine 3.11853.1185 -0.0398 -0.0398 LPE(18:1)+HCOOH(adduct)LPE(18:1)+HCOOH(adduct) 5.46475.4647 -3.6360 -3.6360 LPE(18:2)LPE (18:2) 5.635.63 -3.7641 -3.7641 LPE(20:2)LPE (20:2) 5.64555.6455 -6.8056 -6.8056 LPE(20:2)+HCOOH(adduct)LPE(20:2)+HCOOH(adduct) 5.65915.6591 -0.9271 -0.9271 LPE(18:0)+HCOOH(adduct)LPE(18:0)+HCOOH(adduct) 5.92985.9298 -3.2235 -3.2235 LPE(16:0)LPE (16:0) 5.92985.9298 -0.2460 -0.2460 LPE(18:0)LPE(18:0) 5.94465.9446 -0.5953 -0.5953 LPE(18:0)LPE(18:0) 6.03916.0391 -1.0151 -1.0151 LPE(20:1)+HCOOH(adduct)LPE(20:1)+HCOOH(adduct) 6.05116.0511 -3.3017 -3.3017 eicosapentaenoic acideicosapentaenoic acid 6.50116.5011 -8.9179 -8.9179

쌍별귀뚜라미 제조공정 변화에 따른 대사체 성분 상대량 변화 (원물→단백농축액)Changes in the relative amount of metabolite components according to the change in the manufacturing process of paired crickets (raw material → protein concentrate) CompoundsCompounds RTRT 단백 효소 처리 후 상대 증감율(%)Relative increase/decrease rate after proteolytic enzyme treatment (%) 증가increase uric aciduric acid 1.09891.0989 1.4229 1.4229 tyrosinetyrosine 1.4471.447 3.3893 3.3893 acetyl-glutamic acidacetyl-glutamic acid 1.49151.4915 0.1831 0.1831 dopamine sulfatedopamine sulfate 2.05872.0587 0.6737 0.6737 phenylalaninephenylalanine 2.50142.5014 0.4194 0.4194 methyl sulfotyrosinatemethyl sulfotyrosinate 2.80072.8007 2.6190 2.6190 Di(buta-2,3-dienyl)butane-1,4-diamineDi(buta-2,3-dienyl)butane-1,4-diamine 3.11853.1185 3.8993 3.8993 9,12,13-TriHOME9,12,13-TriHOME 4.25054.2505 6.8072 6.8072 LPE(18:2)LPE (18:2) 5.635.63 3.7878 3.7878 LPE(18:0)+HCOOH(adduct)LPE(18:0)+HCOOH(adduct) 5.76465.7646 0.5967 0.5967 LPE(16:0)LPE (16:0) 5.92985.9298 0.8224 0.8224 LPE(18:0)LPE (18:0) 5.92985.9298 0.0437 0.0437 LPE(18:0)LPE (18:0) 5.94465.9446 1.7158 1.7158 감소decrease xanthinexanthine 6.03916.0391 -0.3080 -0.3080 arabinosylhypoxanthinearabinosylhypoxanthine 2.32592.3259 -2.9869 -2.9869 sulfotyrosinesulfotyrosine 2.74052.7405 -1.6915 -1.6915 LPE(18:1)+HCOOH(adduct)LPE(18:1)+HCOOH(adduct) 5.46475.4647 -3.4880 -3.4880 LPE(20:2)LPE (20:2) 5.64555.6455 -4.0473 -4.0473 LPE(20:2)+HCOOH(adduct)LPE(20:2)+HCOOH(adduct) 5.65915.6591 -0.4408 -0.4408 13-Hydroxyoctadecadienoic acid13-Hydroxyoctadecadienoic acid 5.76465.7646 -0.0520 -0.0520 LPE(20:1)+HCOOH(adduct)LPE(20:1)+HCOOH(adduct) 6.05116.0511 -1.6104 -1.6104 hydroxystearic acidhydroxystearic acid 6.26086.2608 -2.8347 -2.8347 eicosapentaenoic acideicosapentaenoic acid 6.50116.5011 -8.9207 -8.9207

쌍별귀뚜라미 제조공정 변화에 따른 대사체 성분 상대량 변화 (원물→단백농축분말)Changes in the relative amount of metabolite components according to the change in the manufacturing process of paired crickets (raw material → protein-enriched powder) CompoundsCompounds RTRT 단백 농축액 분무건조후
상대 증감율(%)After spray drying of protein concentrate
Relative change rate (%) 증가increase uric aciduric acid 1.4751 1.4751 1.4751 1.4751 tyrosinetyrosine 3.5122 3.5122 3.5122 3.5122 acetyl-glutamic acidacetyl-glutamic acid 0.1952 0.1952 0.1952 0.1952 dopamine sulfatedopamine sulfate 0.6810 0.6810 0.6810 0.6810 phenylalaninephenylalanine 0.5092 0.5092 0.5092 0.5092 methyl sulfotyrosinatemethyl sulfotyrosinate 2.8906 2.8906 2.8906 2.8906 Di(buta-2,3-dienyl)butane-1,4-diamineDi(buta-2,3-dienyl)butane-1,4-diamine 4.0630 4.0630 4.0630 4.0630 9,12,13-TriHOME9,12,13-TriHOME 7.2459 7.2459 7.2459 7.2459 LPE(18:2)LPE (18:2) 3.5750 3.5750 3.5750 3.5750 13-Hydroxyoctadecadienoic acid13-Hydroxyoctadecadienoic acid 0.0482 0.0482 0.0482 0.0482 LPE(18:0)+HCOOH(adduct)LPE(18:0)+HCOOH(adduct) 0.2176 0.2176 0.2176 0.2176 LPE(16:0)LPE (16:0) 0.7824 0.7824 0.7824 0.7824 LPE(18:0)LPE (18:0) 0.1507 0.1507 0.1507 0.1507 LPE(18:0)LPE (18:0) 1.8309 1.8309 1.8309 1.8309 감소decrease xanthinexanthine 1.25611.2561 -0.3013 -0.3013 arabinosylhypoxanthinearabinosylhypoxanthine 2.32592.3259 -3.4250 -3.4250 sulfotyrosinesulfotyrosine 2.74052.7405 -1.5499 -1.5499 LPE(18:1)+HCOOH(adduct)LPE(18:1)+HCOOH(adduct) 5.46475.4647 -3.4568 -3.4568 LPE(20:2)LPE (20:2) 5.64555.6455 -5.0595 -5.0595 LPE(20:2)+HCOOH(adduct)LPE(20:2)+HCOOH(adduct) 5.65915.6591 -0.4346 -0.4346 LPE(20:1)+HCOOH(adduct)LPE(20:1)+HCOOH(adduct) 6.05116.0511 -1.3127 -1.3127 hydroxystearic acidhydroxystearic acid 6.26086.2608 -2.7458 -2.7458 eicosapentaenoic acideicosapentaenoic acid 6.50116.5011 -8.8913 -8.8913

본 발명에 따른 쌍별귀뚜라미 제조공정 변화에 따른 대사체 성분 상대량 변화를 분석하기 위하여, 쌍별귀뚜라미 원물, 착유 및 단백가수분해 처리를 통한 탈지, 단백효소 처리 및 분말화 과정을 거치며 식품에 사용되는 쌍별귀뚜라미 소재의 제조공정별 시료에 대한 대사체 분석을 실시한 결과, 수백가지의 peak로 확인되는 다양한 물질 중 주요한 변화에 영향을 끼친(VIP value >1) 23종의 대사체를 동정하였다. 그 중 쌍별귀뚜라미 원물에 존재하는 인지질(LPE)과 잔틴 성분은 원료 대비 착유공정 및 단백가수분해 효소처리 공정을 거치며 총 대사체 성분 중 차지하는 비율이 14.2%, 2.7%로 각 각 감소하였으며, 티로신, 글루타민산, 페닐알라닌 등의 아미노산과 그 유도체는 총 대사체 성분 함량 중 6.6% 늘어남을 확인했다. 또한 단백가수분해 농축액을 분무건조하여 제조한 수용성 단백분말 에서는 포화지방산인 stearic acid가 3%가량 감소, 앞서 언급한 아미노산과 유도체는 7.2% 증가하는것으로 확인하였다.In order to analyze the change in the relative amount of metabolite components according to the change of the pairwise cricket manufacturing process according to the present invention, the pairwise cricket raw material, milking and degreasing through proteolytic treatment, protease treatment and powdering process are used in food. As a result of metabolite analysis of samples for each manufacturing process of cricket material, 23 metabolites were identified that affected major changes (VIP value >1) among various substances identified as hundreds of peaks. Among them, the phospholipid (LPE) and xanthine components present in the raw material of paired crickets went through the milking process and proteolytic enzyme treatment process compared to the raw material, and the proportion of the total metabolite components decreased to 14.2% and 2.7%, respectively, and tyrosine, It was confirmed that amino acids such as glutamic acid and phenylalanine and their derivatives increased by 6.6% of the total metabolite content. In addition, in the water-soluble protein powder prepared by spray-drying the protein hydrolysis concentrate, it was confirmed that the saturated fatty acid, stearic acid, decreased by 3% and the amino acids and derivatives mentioned above increased by 7.2%.

이상, 본 발명을 바람직한 제조예, 실시예 및 실험예를 통해 상세히 설명하였으나, 본 발명의 범위는 특성 실시예에 한정되는 것은 아니며, 첨부된 특허청구범위에 의하여 해석되어야 할 것이다. 또한, 이 기술분야에서 통상의 지식을 습득한 자라면, 본 발명의 범위에서 벗어나지 않으면서도 많은 수정과 변형이 가능함을 이해하여야 할 것이다.As mentioned above, although the present invention has been described in detail through preferred preparation examples, examples and experimental examples, the scope of the present invention is not limited to the characteristic examples, and should be interpreted by the appended claims. In addition, those skilled in the art will understand that many modifications and variations are possible without departing from the scope of the present invention.

Claims (10)

(1) 쌍별귀뚜라미를 60℃ 내지 80℃의 온도에서 압착하고 착유하여 지방을 제거하고 쌍별귀뚜라미 착유박을 얻는 단계; 및
(2) 단계 (1)에서 얻은 쌍별귀뚜라미 착유박에 단백질 가수분해효소를 이용하여 가수분해하여 가수분해물을 얻는 단계;
(3) 단계 (2)에서 얻은 가수분해물을 원심분리하여 상등액을 얻는 단계;
(4) 단계 (3)에서 얻은 상등액을 농축하고 분무건조하여 분말을 얻는 단계;를 포함하는 것을 특징으로하는 쌍별귀뚜라미 단백질 분말의 제조방법으로,
상기 단계 (2)의 가수분해물을 얻는 단계는 (2)-1 가수분해효소인 플레버자임(Flavourzyme)을 첨가하여 40분 내지 100분 동안 가수분해하는 단계;
(2)-2 상기 단계 (2)-1에서 플레버자임으로 가수분해한 가수분해물에 프로타 맥스(Protamax)를 첨가하여 40분 내지 100분 동안 가수분해하는 단계;
(2)-3 상기 단계 (2)-2에서 프로타맥스로 가수분해한 가수분해물에 알칼레이즈(Alcalase)를 첨가하여 40분 내지 100분 동안 가수분해하는 단계; 및
(2)-4 상기 단계 (2)-3에서 알칼레이즈로 가수분해한 가수분해물에 뉴트라제(Neutrase)를 첨가하여 40분 내지 100분 동안 가수분해하는 단계;를 포함하는 것을 특징으로 하는, 쌍별귀뚜라미 단백질 분말의 제조방법.
(1) pressing and milking the twin-star crickets at a temperature of 60° C. to 80° C. to remove fat and obtain milking meal from the twin-star crickets; and
(2) hydrolyzing the milk meal obtained in step (1) using a proteolytic enzyme to obtain a hydrolyzate;
(3) centrifuging the hydrolyzate obtained in step (2) to obtain a supernatant;
(4) concentrating the supernatant obtained in step (3) and spray-drying to obtain a powder;
The step of obtaining the hydrolyzate of step (2) includes the steps of (2)-1 hydrolyzing by adding a hydrolase, Flavorzyme, for 40 to 100 minutes;
(2)-2 adding Protamax to the hydrolyzate hydrolyzed with flavorzyme in step (2)-1 and hydrolyzing for 40 to 100 minutes;
(2)-3 adding Alcalase to the hydrolyzate hydrolyzed with Protamax in step (2)-2 and hydrolyzing for 40 to 100 minutes; and
(2)-4 adding neutrase to the hydrolyzate hydrolyzed with alkalize in step (2)-3 and hydrolyzing for 40 to 100 minutes; characterized in that it comprises; A method for producing twin-star cricket protein powder.
 삭제delete 제1항에 있어서,
상기 착유박을 얻는 단계(단계 (1))에서 쌍별귀뚜라미를 65℃ 내지 75℃에서 착유하는 것인, 쌍별귀뚜라미 단백질 분말의 제조방법.
The method of claim 1,
In the step (step (1)) of obtaining the milking meal, milking the paired crickets at 65° C. to 75° C., a method for producing a paired-star cricket protein powder.
 제1항에 있어서,
상기 가수분해물을 얻는 단계(단계 (2))에서 이용하는 가수분해효소의 총 농도는 0.10w/w% 내지 1.00w/w%인, 쌍별귀뚜라미 단백질 분말의 제조방법.
The method of claim 1,
The total concentration of the hydrolase used in the step of obtaining the hydrolyzate (step (2)) is 0.10 w/w% to 1.00 w/w%, a method for producing a cricket protein powder by pair.
 삭제delete 삭제delete 삭제delete 삭제delete 제1항의 제조방법으로 제조된 쌍별귀뚜라미 단백질 분말을 함유하는 식품조성물.A food composition containing the double-star cricket protein powder prepared by the method of claim 1. 삭제delete
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JP2009254348A (en) 2008-03-28 2009-11-05 Shingy:Kk Method for producing food and drink of processed larva of bee, and food and drink of processed larva of bee
JP2018502203A (en) 2014-12-31 2018-01-25 インセクト Enzymatic hydrolysis of chitin, hydrolysates, and insects to produce one or more desired products

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JP2009254348A (en) 2008-03-28 2009-11-05 Shingy:Kk Method for producing food and drink of processed larva of bee, and food and drink of processed larva of bee
JP2018502203A (en) 2014-12-31 2018-01-25 インセクト Enzymatic hydrolysis of chitin, hydrolysates, and insects to produce one or more desired products

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