초록 ▼

- 넙치 가공부산물을 원료로 하여 초고압 반응조건에서 효소에 의한 최적 가수분해물 생산 기술을 확립하였으며 이를 마이얄 반응 수식화하여 알러지 저감화되고 항산화,항염증 효능이 우수한 기능성 소재로 개발하고 이를 이용하여 단백질 보충식품 및 향미제 소재로 개발하였음.

- 넙치의 식품 가공 과정 중 생기는 부산물을 이용하여 고압반응기를 이용한 효소 가수분해를 실시하여 얻은 FPH의 분자량 분획과 ribose 당원을 지질로 하여 Maillard 반응할 시 갈변도, DPPH radical 소거활성，FRAP 활성, Histami

- 넙치 가공부산물을 원료로 하여 초고압 반응조건에서 효소에 의한 최적 가수분해물 생산 기술을 확립하였으며 이를 마이얄 반응 수식화하여 알러지 저감화되고 항산화,항염증 효능이 우수한 기능성 소재로 개발하고 이를 이용하여 단백질 보충식품 및 향미제 소재로 개발하였음.

- 넙치의 식품 가공 과정 중 생기는 부산물을 이용하여 고압반응기를 이용한 효소 가수분해를 실시하여 얻은 FPH의 분자량 분획과 ribose 당원을 지질로 하여 Maillard 반응할 시 갈변도, DPPH radical 소거활성，FRAP 활성, Histamine 방출정도，β-hexosaminidase 방출정도，Cholesterol Micelle 저감 활성이 최대가 되는 GFPH의 생산 최적 MR 반응 조건을 RSM분석에 의하여 확립하였음.

- GFPH는 향미가 우수하고 열적변화에 안정적이며 친수성이 강한 특성을 가지며 안전한 것으로 판단되었으며 GFPH와 향미전구체，부재료를 이용한 단백질 보충제 및 향미제를 개발하였으며 최종 제품은 저장기간 중 안정한 것으로 판단되었음.

- GFPH는 알러지 반응이 유의적으로 감소하였으며 염증 신호전달 경로 억제에 위한 항염증 활성이 우수하였으며, 특히 항산화활성이 뛰어나 이를 바탕으로 간 보호 활성을 확인한 결과, FPH에 비하여 간 보호 활성이 크게 증가함을 확인하였음.

(출처 : 보고서 요약서 3p)

Abstract ▼

IV. Results
The results of the research and development for each part of the project are summarized as follows.
Part I : Development of production techniques and materialization for functional GFPH with Maillard reaction and enzymatic hydrolysis with ultra high pressure
-High hydrostatic p

IV. Results
The results of the research and development for each part of the project are summarized as follows.
Part I : Development of production techniques and materialization for functional GFPH with Maillard reaction and enzymatic hydrolysis with ultra high pressure
-High hydrostatic pressure enzymatic hydrolysis was applied to the flatfish byproducts (FFP) in order to develop the functional food materials. Proximate compositions of FFP were determined and its crude protein content, which could be converted to bioactive peptide, was 20.4 %. The degree of hydrolysis (DH) of FFP hydrolysates were investigated depending on the process parameters such as pressure, temperature, [substrate]/[enzyme] ([S]/[E]) ratio, pH and type of enzyme. The results showed that temperature, s/e ratio and pH are major independent parameters to optimize a low-cost commercial process under 100 MPa fixed pressure using Protamex™ protease. Response surface methodology (RSM) was applied to study the effects of three major high hydrostatic pressure enzymatic hydrolysis parameters and the RSM statistical analysis results showed the hydrolysis parameters for optimal DH : [S]/[E] ratio of 50.65 (w/w), pH 6.89 and temperature of 56.97 ℃. Under the optimal condition, the DH of FFP was predicted as 32.09%. To enhance the bioavailability, FFP hydrolysates was processed through a series of ultrafiltration membrane with molecular weight cut-off (MWCO) 10， 5 and 3 kDa. Four types of permeates including below 3 kDa，3-5 kDa，5-10 kDa and over 10 kDa were obtained and comparative analysis of amino-acid compositions was conducted. These results suggest that high hydrostatic pressure enzymatic hydrolysis is effective processing method for utilization of FFP as functional food materials.

- We investigated the optimal conditions of the Maillard reaction (MR) with ribose using response surface methodology to reduce the allergenicity of its protein. A 3-factored and 5-leveled central composite design was used, where the independent variables were substrate (ribose) concentration (X₁，%), reaction time (X₂, min), and pH (X₃)，while the dependent variables were browning index (Y₁，absorbance at 420 nm)，DPPH scavenging (Y₂, EC50 mg/mL), FRAP (Y₃, mM FeSOVmg extract) and b-hexosaminidase release (Y₄, %). The optimal conditions were obtained as follows: X₁, 28.36%； X₂, 38.09 min; X₃, 8.26. Maillard reaction products of fish protein hydrolysate (MFPH) reduced the amount of nitric oxide synthesis compared to the untreated FPH, and had a significant anti-allergy effect on b-hexosaminidase and histamine release, compared with that of the FPH control. We concluded that MFPH, which had better antioxidant and anti-allergy activities than untreated FPH, can be used as an improved dietary source.

- Physiochemical and nutritional characteristics of glycated-fish protein hydrolysates (GFPH) were investigated. From four GFPHs with different molecular weight, the production yield and browning index were high in GFPH from under 5 kDa. Slightly higher palatability of sensory evaluation was noted in GFPHs from low molecular weight when compared with high molecular weight. A dynamic characteristics measured storage modulus (G') and loss modulus (G" ) increased as the frequency increase and G' presented higher value than G" . As concentration of GFPH decrease, values for the surface tensions were increased but density did not show consistency. The emulsifying capacity of GFPH improved from 2.41% of FPH to 28.79%, emulsifying stability and the solubility of GFPH were higher than those of FPH. Foaming stability was also increased by glycation of fish protein hydrolysates. Burnt smell and fishy smell were not significantly different among all samples. The palatability was correlated with fishy smell and meaty smell even though there were no significances between GFPH and FPH. GFPH has a potential application as an additives and functional materials.

- Reaction flavor based on Maillard reaction (MR) products with flatfish byproducts hydrolysates was developed and characterized its sensory properties and identified volatile compounds using GC/MS and e-nose analyzer. Glycated flatfish protein hydrolysates (GFPH) based flavors were prepared using MR with various concentrations of ingredients and optimized formula of production (GFPH: 0.5 g, DL—methionine: 0.025 g, lecithin: 0.0125 g, L-cysteine 0.05 g and thiamine: 0.01 g) was established based on evaluating sensory properties. GFPH based flavor was analyzed using GC/MS and total 64 volatile compounds were identified (13 acids, 12 alcohols, 10 aldehydes, 7 ketones: 3 pyrazines, 3 sulfur-containing compounds, 3 furans and 13 Miscellaneous). Among them, benzaldehyde and 3-methyl-butanal are considered as having positive role in flavor sensory properties. Furthermore, volatile flavors such as ethyl isobutyrate and phenylethanol
were identified using mass spectrometry based e-nose analysis. We It is concluded that GFPH based flavor has strong potential in the field of flavor application in food industry.

- GFPH based protein supplements were formulated with soy protein isolates, defatted whole milk powder, cocoa powder, chocolate chips, polydextrose, maltodextrin (non digestible), Vitamin mix, Vit. C. stevia (enzyme treated) and silicon oxide and flavored with dried apple powder. Products with optimized formula were stable during storage conditions.

- To confirmed the general toxicity of GFPH, we tested a single- and 14-day repeated dose oral toxicity study. In the oral toxicity study, the single oral dose of the GFPH at 1000 mg/kg did not produce mortality or abnormal lesions in the internal organ of rats. The results of a 14-day orally repeated dose showed that the GFPH gave no adverse effects at dosages of 1000 mg/kg in rats in the study.

Part II : Verification of efficacy on allergy reduction, antioxidative and anti-inflammation activity of glycated FPH

- The anti-allergy effect of Maillard reaction products (MRPs) with ribose was investigated. Fish protein hydrolysate (FPH) with ribose at a molar ratio of 1:1 was dissolved in distilled water (the final concentration of substrate was 28%), adjusted to pH 8.26, and incubated at 121℃ for 38 min. Maillard reaction products (MRPs) of fish protein hydrolysate (FPH) reduced nitric oxide synthesis compare with untreated FPH. And anti-allergy effect of MRPs, which measured by β-hexosaminidase and histaminerelease, was significantly increased compared with that of FPH control. However, thereis no significant difference in cholesterol reduced effect. Therefore, we concluded that MRPs of FPH (GFPH) had antioxidant and anti-allergy activities.

-The anti-inflammatory effects of glycosylated fish byproduct protein hydrolysate (GFPH) and their anti-inflammatory mechanisms were elucidated in lipopolysaccharide (LPS) - stimulated RAW264.7 mouse macrophage. GFPH suppressed LPS-induced production of nitric oxide (NO) and prostaglandin E₂ (PGE₂) and expression of inducible nitricoxide synthase(iNOS) and cyclooxygenase-2 (COX-2) with dose dependent manner. Enzyme-linked immunosorbentassay ELISA) kit clearly demonstrated that GFPH significantly reduced the productions of pro- inflammatory cytokines such as, interleukin(IL)-6, interleukin (IL)-Iβ and tumor necrosis factor (TNF)-α, and monocyte chemoattractant protein (MCP)-1. Moreover, GFPH reduced nuclear factor kB (NF-kB) and mitogen-activated protein kinases (MAPKs) activation.

- Molecular mechanisms of hepatic protection effect of GFPH in vitro model were further investigated. GFPH protected liver HepG2 cells against t-BHP-induced oxidative stress with enhanced glutathione (GSH) synthesis in vitro. Furthermore, we showed that GFPH induced upregulation of phase II enzyme expression, such as that of HO-1 and γ-GCL, via nuclear translocation of Nrf2 and phosphorylation of ERK. Taken together, these results indicate the potential of GFPH for use as a functional food ingredient with improved rheological and antioxidative properties.

- In order to determine whether t-BHP-induced oxidative stress can be prevented by GFPH, we performed an in vivo study. t-BHP treated groups showed significant reductions of serum AST and ALT. And we measured various enzyme activity related with antioxidant effect such as GSH level, CAT and SOD activities in liver tissue. t-BHP treated group also significantly decreased the CAT and SOD activities compared with control group. Although FPH and GFPH treated group fully recovered CAT activity to the control, GFPH remarkably enhanced SOD enzyme activities than FPH. Finally, we described that GFPH had more hepatic protection activity rather than FPH, while both FPH and GFPH significantly increased antioxidant relevant enzyme activities even when exposed to t-BHP-induced oxidative damage in liver.

(출처 : SUMMARY 12p)

목차 Contents

• 표지 ... 1
• 제 출 문 ... 2
• 보고서 요약서 ... 3
• 요약문 ... 5
• SUMMARY ... 11
• CONTENTS ... 17
• 목차 ... 18
• 제 1 장 연구개발과제의 개요 ... 19
• 제 1 절 연구개발의 필요성 ... 19
• 제 2 절 연구개발의 중요성 ... 23
• 제 3 절 연구개발범위 ... 24
• 제 2 장 국내외 기술개발 현황 ... 26
• 제 1 절 관련기술 국외 기술 동향 ... 26
• 제 2 절 관련기술 국내 기술 동향 ... 29
• 제 3 절 관련기술 특허 동향 ... 31
• 제 3 장 연구개발 수행 내용 및 결과 ... 35
• 제 1 절 연구 재료 및 방법 ... 35
• 1. 고압처리 및 마이얄 반응을 이용한 알러지 저감화 어류단백 가수분해물 생산기술 개발과 이를 이용한 기능성 식품소재 개발 ... 35
• 2. 당화 FPH의 알러지 저감효능, 항산화 효능, 항염증 효능 및 심혈관 보호 효과 검증 기술 개발 ... 64
• 제 2 절 연구결과 ... 84
• 1. 고압처리 및 마이얄 반응을 이용한 알러지 저감화 어류단백 가수분해물 생산기술 개발과 이를 이용한 기능성 식품소재 개발 ... 84
• 2. 당화 FPH의 알러지 저감효능, 항산화 효능, 항염증 효능 및 심혈관 보호 효과 검증 기술 개발 ... 190
• 제 4 장 목표달성도 및 관련 분야에의 기여도 ... 249
• 제 1 절 연구개발 목표 달성도 ... 249
• 제 2 절 관련분야에의 기여도 ... 250
• 제 5 장 연구개발 성과 및 성과활용 계획 ... 252
• 제 1 절 연구개발 성과 ... 252
• 제 2 절 성과 활용 계획 ... 254
• 제 6 장 참고문헌 ... 256
• 끝페이지 ... 265