초록 ▼

○ 연구결과
1.주관기관:단백질 회수방법 최적화 및 고부가가치 제품 산업화
가.단백질 회수방법 정립
(1)케이크 여과법(플라스틱 및 삼베망 각 2겹)
(2)수율이 2배 높고 겔 특성 효율적인 커팅법(8분,15℃,100%)정립
나.회수단백질 적정 첨가 수준 설정
(1)커팅육 크래미 20%,어묵 10% 연육 대체 가능
다.가수분해물 및 축육 함유 섬유상 연제품 개발
(1)가수분해물 게맛살 및 소시지 1.0%,어묵 0.4% 첨가 가능
라.MDCM 추출 생리활성 펩티드 특성 구명 및 적정 첨

○ 연구결과
1.주관기관:단백질 회수방법 최적화 및 고부가가치 제품 산업화
가.단백질 회수방법 정립
(1)케이크 여과법(플라스틱 및 삼베망 각 2겹)
(2)수율이 2배 높고 겔 특성 효율적인 커팅법(8분,15℃,100%)정립
나.회수단백질 적정 첨가 수준 설정
(1)커팅육 크래미 20%,어묵 10% 연육 대체 가능
다.가수분해물 및 축육 함유 섬유상 연제품 개발
(1)가수분해물 게맛살 및 소시지 1.0%,어묵 0.4% 첨가 가능
라.MDCM 추출 생리활성 펩티드 특성 구명 및 적정 첨가 수준 설정
(1)펩티드는 항산화성보다 ACE 저해능 탁월
(2)펩티드 게맛살 1.0% 첨가시 간 보호 활성 기여
마.펩티드 및 축육 함유 섬유상 연제품 개발(세계 최초)
(1)킹크랩 개발시 커팅 폐계 가슴살 10%까지 연육 대체 가능
(2)소시지에 펩티드 첨가시 조직 개선에는 ISP보다 감자전분 효과 높음
(3)어묵에 동결건조 펩티드 첨가시 POV 낮으나 겔 및 관능 특성 저하
(4)대부분의 개발 연제품 및 소시지류 공히 냉장온도에서 4주 이상 유통 가능
2.협동기관:회수단백질을 이용한 조미소재 및 생리활성 펩티드 특성 구명
가.회수단백질의 수율은 56.7%
나.Protamex가수분해물의 최적조건:E/S 0.06,pH 7.0,온도 43℃
다.2단 최적 단백질 분해효소:Bromelain
라.2단 가수분해물 제조의 최적 조건:E/S 3.56,pH 5.06,온도 50.2℃
마.유리 아미노산의 함량:1단가수분해<2단 가수분해물바.이취제거 Maillard반응을 위한 최적 ingredient조성의 최적화
사.DPPH 라디칼 소거능:2단 가수분해물<이취제거 Maillard반응물
아.DPPH 라디칼 소거능 펩티드:KLGXX,GGXXX,KGGGX,XGXRY,XQPQX,XQPQV
자.ACE 저해 펩티드:YAK(IC50=0.197uM),LTK(IC50=0.171uM),LY(IC50=2.344uM), TYP(IC50=22.585uM),FQNPD(IC50=1.908uM)
차.합성 펩타이드의 세포독성:cell-line에서 검출되지 않음
카.LY,TYP,FQNPD;collagenasetypeI저해
타.합성 펩티드의 돌연변이원성:확인되지 않음

Abstract ▼

A. Managerial institution (Gyeongnam national university of science and technology)
The objectives of this study were to develop of optimum method to extract of recovered protein derived using spent laying hens and then apply it to produce valuable products.
1. The first year of the experiment

A. Managerial institution (Gyeongnam national university of science and technology)
The objectives of this study were to develop of optimum method to extract of recovered protein derived using spent laying hens and then apply it to produce valuable products.
1. The first year of the experiments
1) Recovery protein was collected due to pH after ISP addition and then cake filteration.
After recovery protein was collected, gel characteristics, folding test and texture properties of recovery protein were measured. Generally, gel characteristics, folding test and texture properties were lowered but recovery protein was collected due to pH only had a better value in gel characteristics.
2) Redness was lowered when salt was added to recovery protein, however, folding test was improved.
3) Lightness, whiteness and folding test was improved when recovery protein showed better properties, also, salt addition to recovery protein improved in gel properties.
4) Before and after cake filteration using 5 and 1 mm of sieve and 1 mm stainless filter provided negative effects to recovery proteins, however, using 2 layers of plastic and 2 layers of hemp cloth to recovery protein, the recovery protein had better properties in folding test and gel characteristics.
5) The addition of 2, 4 and 6 layers of filter did not affect on the gel characteristics of recovery proteins when salt was added to each treatment, in contrast, gel characteristics of recovery proteins were not improved when salt was not added to recovery proteins and it is because cross-bridges were not completely built up when salt was not added.
6) Lightness, whiteness, folding test, breaking force and jelly strength of FA Pollack had higher values, in contrast, redness, yellowness and shear force were not higher as compared to those of KA and FA, and the addition of salt to Pollack provided a positive effect on gel characteristics of Pollack, which is lower in grade.
7) The addition of salt to recovery proteins significantly improved the values of breaking force, gel force, folding test and texture properties, however, the value of deformation was decreased.
8) Higher grade of recovery protein showed higher values of lightness, whiteness, folding test, breaking force and deformation but had lower values in redness and yellowness.
No significant difference was determined in shear force.
9) Optimum cutting was conducted when 8 min per Pollack was applied and over 8 min cutting generated the protein denature, thereby, the low level of gel characteristics was determined. The sheet formation was important when cutting was applied to the pollacks. Before the formation of sheet, all materials must be filtered through a 1 mm sieve. Also, cutting method had a high recovery ratio, which was not found in pH and cake filtration methods.
10) Better gel characteristics were determined when cutting method was applied to pollacks as compared to that of pH method and shear force and folding test were improved when cutting time was increased. Optimum cutting time was 8 min and it was determined due to the consideration of protein denature of 12 min cutting.
11) Cremis from all treatments showed lower values in lightness, moisture content, TBARS and VBN as compared to those of control. Cremis in T2 had high values in whiteness, breaking force, deformation and gel strength of gel characteristics and hardness of texture properties and Cremis from T3 treatment were high in salt soluble protein concentration, cohesiveness of texture properties and shear force. In sensory evaluation, no significant differences were determined, thereby, as the consideration of economical, quality and yield properties, recovery proteins derived using a cutting method may be replaced up to 15% and it may work with normal qualities.
2. The second year of the experiments
1) To determine the effects of the recovery protein addition derived using a cutting method on the quality and storage time of Cremis, spent layer recovery protein (SLRP) was added to Pollack and 0 (C), 10 (T1), 20 (T2), 30 (T3) and 40% (T4) of pollack was replaced and Cremis were manufactured and then stored at 10℃ for 4 weeks of storage. The addition of SLRP increased the yellowness in color, gel strength and texture properties, however, whiteness was decreased. No significant differences were determined in sensory evaluation, thereby, replacement of 20% Pollack by SLRP maybe economically acceptable and up to 30% replacement maybe considerable.
2) To determine the effects of the recovery protein addition manufactured using a cutting method on the quality and storage time, spent layer recovery protein (SLRP) was added to boiled fish pastes and 0 (C), 10 (T1), 20 (T2), and 30% (T3) of pollack was replaced and boiled fish pastes were processed. Each paste was then stored at 10℃ for 6 weeks. The addition of SLRP affected the quality of boiled fish pastes and improved the lightness and yellowness, however, decreased the values in redness and whiteness. Moreover, the addition of SLRP showed positive effects on gel strength and texture properties but no significant difference was observed due to sensory
panels. Therefore, the addition of SLRP up to 10% may be considerable to process boiled fish pastes.
3) To determine the effects of the MDCM hydrolysates addition up to 0 (C), 0.5 (T1), 1.0 (T2) and 1.5% (T3) on the quality and storage time of imitation crab stick, imitation crab sticks were manufactured and then stored at 10℃ for 4 weeks. Lightness and whiteness of imitation crab stick were decreased but had high values in gel characteristics when more MDCM hydrolysates was added. In addition the MDCM addition was not significantly affected on texture properties of imitation crab sticks but showed high values in brittleness, gumminess and chewiness and showed low especially in POV. No significant differences were not determined, thereby, the imitation crab sticks contained 20% of recovery proteins and 1.0% of MDCM
hydrolysates may positively be stored up to 4 weeks of storage.
4) To determine the effects of the MDCM hydrolysates addition up to 0 (C), 0.4 (T1), and 0.8% (T2) on the quality and storage time of boiled fish pastes, boiled fish pastes were manufactured and then stored at 10℃ for 6 weeks. The whiteness was decreased and gel characteristics were increased when MDCM hydrolysates was added and then processed to boiled fish pastes. However, no significant differences were observed in texture properties. Positive effects were determined in DPPH when MDCM hydrolysates was added to boiled fish pastes. No significant differences were observed in sensory evaluation, thereby, boiled fish pastes contained 20% of recovery proteins and 0.4% of MDCM hydrolysates may positively be stored up to 6 weeks of storage.
5) To determine the effects of the MDCM hydrolysates addition up to 0 (C), 1 (T1), 2 (T2), 3 (T3), 0.05% vitamin C (T4) and ascorbic acid (T5) on the quality and storage of sausages, sausages were processed and then stored at 10℃ for 4 weeks. The lightness, whiteness and gel characteristics of sausages were increased when more MDCM hydrolysates was added but no significant differences were observed in texture properties. Such effects were influenced on the quality of sausages and significant differences were determined due to panels when sausages were stored after 2 weeks of storage. Therefore, up to 1.0% of MDCM hydrolysates addition to sausages may positively be considerable to manufacture sausages using MDCM hydrolysates.
3. The third year of the experiments
1) Second MDCM hydrolysates had low in DPPH, xanthine oxidase inhibitory activity, superoxide scavenging activity and hydroxyl radical scavenging activity. However, ACE inhibitory property related to liver functions was significantly increased as compared to that of control.
2) To determine the effects of the double distilled second MDCM hydrolysates addition up to 0, 1.0 and 1.5% on the quality and storage time of imitation crab sticks, imitation crab sticks were manufactured and then stored at 10℃ for 6 weeks. pH of imitation crab sticks was significantly increased, in contrast, moisture content was decreased as the double distilled second MDCM hydrolysates concentration was increased. In addition, the addition of double distilled second MDCM hydrolysates concentration in imitation crab sticks CIE L*, a* and b*, chroma and hue values were increased but sometimes decreased depending on the concentration of second MDCM hydrolysates. Also, VBN, TBARS and POV and breaking force, gel strength and jelly strength were increased due to second MDCM hydrolysates concentration was increased. However, no significant differences were observed due to the panels and determined in DPPH, superoxide radical scavenging and hydroxyl radical scavenging properties. Only ACE inhibitory property was significantly differed in the imitation crab sticks of 1.0% second MDCM hydrolysates treatment.
3) To determine the effects of the recovery protein addition up to 8 (T1) and 17% (T2) on the quality and storage time of King-crabs, king clicks were manufactured and then stored at 10℃ for 3 weeks. The moisture content of two different treatments was increased as storage time was increased and low TBARS values were determined in king clicks derived from T2 as compared to that of T1. The yellowness and redness was increased but decreased respectively when recovery proteins were replaced up to 8 and 17%. Whiteness of king click was significantly decreased.
Breaking force and jelly strength of king clicks were significantly affected due to the addition of recovery proteins rather than storage time and such effects were not influenced on sensory evaluation.
4) To determine the effects of the second MDCM hydrolysates, ISP and potato starch addition, cooked pork sausages were manufactured and then stored at 10℃ for 4 weeks and the quality effects of each sausage were determined on 0, 2 and 4 weeks of storage. The moisture content was the highest and it was 67.67–67.90% when 2.0% ISP was added to sausages but significantly high TBARS, VBN and POV values were determined. The addition of ISP to sausages showed effects on texture properties of sausages and the ISP influenced on the deformation, gel strength and jelly strength and had positive effects on hardness, gumminess and adhesiveness. The addition of second MDCM hydrolysates, ISP and potato starch to cooked pork sausages showed negative effects on odor, flavor and tenderness but no significant difference was observed in overall acceptability at 4 weeks of storage.
5) To determine the effects of the freeze dried and second MDCM hydrolysates addition up to 1 and 3% on the quality and storage time of boiled fish pastes, boiled fish pastes were manufactured and then stored at 10℃ for 4 weeks. The quality of boiled fish pastes was determined on 0, 2 and 4 weeks of storage. The moisture content of boiled fish pastes was 76.33–76.80%. The highest moisture content was determined when 3% of second MDCM hydrolysates was added and it is 76.80%. The CIE a* and b* values were increased and moisture content, shear force, TBARS and VBN values were also increased, in contrast, pH and POV were decreased as second MDCM hydrolysates values were increased up to 3% to boiled fish pastes. In addition, gel characteristics and texture properties were decreased when second MDCM hydrolysates addition was increased from 1 to 3%, thereby, such effects were affected to the values derived from sensory panels. Control showed higher sensory values as compared to those of treated groups.
B. Cooperative institution (Gyeongsang national university) The objectives of this study were to investigate food and bioactive functionalities of the recovered protein hydrolysates from mechanical deboned chicken meat (MDCM), and a potential application.
1. The first year of the experiments
1) The yield of recovered protein from MDCM was 56.7% by alkaline pH shifting process. The optimum protease was Protamex in the first step of hydrolysis. The optimum conditions for hydrolysis were 0.06 for ratio of protease to MDCM, 7.0 for pH, 43℃ for temperature, and 6 hr for reaction time, respectively. Free amino acid was 428.27±1.1 mg/100 mL in Protamex hydrolysate. The major minerals were phosphate, calcium, sodium and potassium. The solubility of Protamex hydrolysate increased up to pH 8.0, and did not affected by NaCl concentration. Emulsifying activity index (EAI) of Protamex hydrolysate was higher than that of gluten, but significantly lower than those of bovine albumin and soy protein isolate. The emulsifying stability index (ESI) of Protamex hydrolysate was also much lower than other non-muscle protein. The results suggested that Protamex hydrolysate from MDCM did not have a potential properties for seasoning. Protamex hydrolysate was needed a two step hydrolysis, enzymatic and chemical modification for seasoning source.
2) The conditions for plastein reaction were 0.7 mL for deamidated gluten 0.1 mL for Protamex hydrolysate, 0.4 mL of buffer (pH 6.0), and 0.4 mL for papain.
Polymerization of plastein material was 77.1%. The minimum TCA soluble index was showed in 0.9 mL for deamidation gluten, 1.5 mL for Protamex hydrolysate, 0.4 mL for buffer (pH 6.0), 0.25 mL for papain, respectively. Glutamic acid was composed of 22.0% in total amino acids. Free amino acid of plastein material was 3226.7 mg/100g.
Glutamic acid as taste compound was 3.5% in total free amino acid. Solubilities were not affected by NaCl concentration and pH. EAI, ESI, water and fat adsorption of plastein material were higher than those of bovine serum albumin but much lower than those of non-muscle protein.
2. The second year of the experiments
1) The contents of soluble compounds did not depend on proteases such as Alcalase, Neutrase, Flavourzyme and bromelain, significantly (p<0.05). However, DPPH radical scavenging activity was greatly increased in bromelain hydrolysate. Optimum conditions for 2 steps hydrolysis by bromelain were 3.56 for ratio of Protamex hydrolysate to protase, 50.2℃ for reaction temperature and 5.06 for pH. DPPH radical scavenging activity of optimum hydrolysate showed 88.62%. Optimum ingredients were 95.2% Protamex hydrolysate, 1.9% chicken smoking solution, 1.0% chicken oil, 0.5% hydrolysated vegetables protein, 0.5% glucose, 0.5% xylose, and 0.5% onion liquid for Maillard reaction to remove chicken odor. Optimum temperature and time were 95℃ and 50 min, respectively. Free amino acid content of two step hydrolysate and deodoring hydrolysate were 678.5±1.5 mg/100 g and 683.2±6.9 mg/100 g, respectively.
Free amino acid was increased up to 1.58 fold compared to one step hydrolysate.
Sodium in two step and deodoring hydrolysate was greatly increased, whereas potassium and phosphate were decreased. Cadnium was not detected in both hydrolysate. Hydrolysate of 500 Da composed of 43.4% of total two step hydrolysate, while hydrolysate of 260 Da composed of 69.1% in deodoring hydrolysate. DPPH radical scavenging activities (EC₅₀) of the two step and deodoring hydrolysate were 13.0 mg/mL and 5.04 mg/mL. Angiotension enzyme inhibition (IC₅₀) of the two step and deodoring hydrolysate were 0.032 mg/mL and 0.018 mg/mL, respectively.
Fe-chelating and reducing abilities were high in deodoring hydrolysate compared to two step hydrolysate. Both hydrolysates showed antioxidant effect against linoleic acid. The results suggested that the two step hydrolysate from recovered MDCM protein had a potential application as angiotension enzyme (ACE) inhibitor.
2) The peptides were purified with ion-exchange, size exclusion and reverse-phase chromatography from two step hydrolysate of the recovered MDCM protein to identify ACE inhibitor and DPPH radical scavenger. Amino acid sequences of the purified peptides were K(G)L(G)G(S,L)XX, GGXXX, KG(L,Q)G(Q)G(E)X, XG(L,P)XR(Q)Y, XQPQX and XQPQV for DPPH radical scavenger. Amino acid sequences for ACE inhibitor were YAK, LTK, LY, TYP and FQNPD, and their IC₅₀ values were 0.197 uM, 0.171 uM, 2.344 uM, 22.585 uM and 1.908 uM, respectively.
3. The third year of the experiments
1) Two step and deodoring hydrolysate did not show the toxicity by 200 ug/mL against a normal liver and RAW 264.7 cell line. In addition, the toxicities against a normal liver cell line did not detect in the synthetic peptides. Two step and deodoring hydrolysates showed the protective effect of 80% and 90% against liver cell line treated by ethanol treatment, while synthetic peptides did not show the protective effect. Also, Hydrolysates and synthetic peptides did not reduce a number of AGS and HT 29 cell. The production of NO was not influenced with all hydrolysates. LY and TYP inhibited collagenase type I in the 12.5 fold concentration of protease, while FQNPD inhibited collagenase activities by 60.9% at the same concentration.

목차 Contents

• 표지 ... 1
• 제 출 문 ... 2
• 요 약 문 ... 3
• SUMMARY ... 8
• CONTENTS ... 15
• 목 차 ... 19
• 제1장 연구개발과제의 개요 ... 22
• 제1절 연구개발의 목적 ... 22
• 제2절 연구개발의 필요성 ... 22
• 제3절 연구개발의 범위 ... 23
• 1. 주관기관 과제: 단백질 회수방법 최적화 및 고부가가치 제품 산업화 ... 23
• 2. 협동기관 과제: 회수단백질을 이용한 조미소재 및 생리활성 펩티드 특성 구명 ... 24
• 제2장 국내외 기술개발 현황 ... 26
• 제1절 국내·외 관련분야에 대한 기술개발 현황 ... 26
• 제2절 연구결과가 국내·외 기술개발 현황에서 차지하는 위치 ... 30
• 제3장 연구개발수행 내용 및 결과 ... 31
• 제1절: 단백질 회수방법 최적화 및 고부가가치 제품 산업화 ... 31
• 1. 단백질 회수방법 최적화(1차년) ... 31
• 가. ISP 첨가 후 케이크 여과하여 pH 조절법으로 회수한 단백질의 특성 비교 ... 31
• 나. pH 조절법으로 회수한 단백질에 소금 첨가 효과 규명 ... 34
• 다. 연육 종류 등급 간에 소금 첨가 효과 규명 ... 36
• 라. 케이크 여과법 여과 규격 및 회수단백질에 소금 첨가 효과 규명 I ... 38
• 마. 케이크 여과법 여과 규격 및 회수단백질에 소금 첨가 효과 규명 II ... 41
• 바. 명태 연육 종류 등급 간에 소금 첨가 효과 규명 ... 44
• 사. pH 조절법으로 회수한 단백질에 소금 첨가 효과 추가 규명 ... 47
• 아. 명태 연육 등급 간에 소금 첨가 효과 규명 ... 49
• 자. 커팅법의 커팅조건 정립 비교 ... 51
• 차. 커팅법과 pH 조절법 후 케이크 여과하여 두 방법 간 특성 규명 ... 54
• 카. 단백질 회수 방법 간 크래미 현장시제품의 특성과 유통기한 설정 ... 56
• 2. 회수단백질을 이용한 조미소재 함유 제품 개발(2차년) ... 69
• 가. 커팅법으로 제조한 폐계육의 명태연육 대체 비율에 따른 현장시제 크래미의 품질 특성 및 유통기한 설정 ... 69
• 나. 커팅법으로 제조한 폐계육의 연육 대체 비율에 따른 현장시제 유탕형 어묵의 품질특성 및 유통기한 설정 ... 82
• 다. MDCM 추출 조미소재를 첨가한 현장시제 게맛살의 품질 특성 및 유통기한 설정 ... 93
• 라. MDCM 추출 조미소재를 첨가한 유탕형 어묵의 품질 특성 및 유통기한 설정 ... 104
• 마. MDCM 추출 조미소재를 첨가한 소시지의 품질 특성 및 유통기한 설정 ... 115
• 3. 회수단백질을 이용한 생리활성 펩티드 함유 제품 개발(3차년) ... 127
• 가. 회수단백질의 생리활성 검증 ... 127
• 나. 회수단백질을 이용한 생리활성 펩티드 함유 게맛살의 개발 ... 131
• 다. 회수단백질을 이용한 킹크랩의 개발 ... 149
• 라. 회수단백질을 이용한 생리활성 펩티드 함유 소시지의 개발 ... 159
• 마. 회수단백질을 이용한 생리활성 펩티드 함유 어묵의 개발 ... 175
• 제2절 회수단백질을 이용한 조미소재 및 생리활성 펩티드 특성 구명 ... 186
• 1. 회수단백질을 이용한 조미소재 특성 구명(1차년) ... 186
• 가. 발골계육 회수단백질을 이용한 효소 가수분해물의 특성 ... 186
• 나. Plastein 반응에 의한 회수단백질 가수분해물의 특성 변화 ... 203
• 2. 회수단백질을 이용한 생리활성 펩티드 특성 구명(2차년) ... 218
• 가. MDCM 회수단백질 2단 가수분해물의 생리활성 ... 218
• 나. MDCM 회수단백질로 제조한 2단 가수분해물의 생물활성 펩티드의 정제와 활성 ... 239
• 3. 합성 펩티드의 안전성 검증(3차년) ... 256
• 가. MDCM 회수단백질로 제조한 2단 가수분해물과 합성 펩티드의 안전성 ... 256
• 제4장 목표달성도 및 관련분야에의 기여도 ... 268
• 제1절 연구개발 목표의 달성도 ... 268
• 1. 연구성과 달성도 ... 268
• 2. 연도별 연구개발목표의 달성도 ... 269
• 제2절 관련분야의 기술발전에의 기여도 ... 272
• 1. 기술적 측면 ... 272
• 2. 경제․산업적 측면 ... 272
• 3. 산업․-문화적 측면 ... 272
• 제5장 연구개발 성과 및 성과활용 계획 ... 273
• 제1절 연구개발 성과 ... 273
• 제2절 성과활용 계획 ... 276
• 1. 실용화․산업화 계획 ... 276
• 2. 교육․-지도․.홍보 등 기술 확산 계획 ... 276
• 3. 특허․논문 등 지식재산권 확보 계획 ... 276
• 4. 추가연구․타연구에 활용 계획 ... 277
• 제6장 연구개발과정에서 수집한 해외과학기술정보 ... 277
• 제7장 참 고 문 헌 ... 278
• 끝페이지 ... 280