[보고서]전분 활성 효소를 이용한 기능성 탄수화물 신소재 개발

윤미라

전분 활성 효소를 이용한 기능성 탄수화물 신소재 개발
Developing of functional carbohydrates using carbohydrate active enzymes 원문보기

보고서 정보
주관연구기관	국립식량과학원
연구책임자	윤미라
참여연구자	안승현 , 최임수 , 오세관 , 이정희 , 천아름 , 그외 다수
보고서유형	최종보고서
발행국가	대한민국
언어	한국어
발행년월	2012-01
과제시작연도	2011
주관부처	농촌진흥청
사업 관리 기관	농촌진흥청 Rural Development Administration
등록번호	TRKO201200009936
과제고유번호	1395022286
DB 구축일자	2013-04-18

초록 ▼

1. 제1세부과제 : 쌀 전분의 효소적 맞춤형 구조변형에 적합한 유전자원 탐색 및 품종 선발
가. 기능성 전분 소재 개발을 위한 쌀 전분 구조 특성 차이 분석
(1) 품종별 전분 아밀로오스 함량과 아밀로펙틴 측쇄 사슬 길이 분포 비교
(2) 쌀 배유 층위별 전분 특성 변이
(3) 주요 유전자원의 아밀로오스 함량에 따른 아밀로펙틴 중합도 비교
나. 기능성 탄수화물 소재 적합 품종 탐색과 쌀 전분 구조 특성과의 상관성
(1) 주요 품종별 환형아밀로오스 제조 및 수율 비교
(2) 환형아밀로즈 제조 적성과 쌀 전분 구조 특성과의 연관성 검토
2. 제2세부과제 : 전분활성효소를 이용한 고구마의 기능성 탄수화물 신소재 개발
가. 기능성 전분생산을 위한 고구마 품종 선발
(1) 고구마 품종별 고구마 전분 특성비교
(2) 고구마 품종별 원료 전분 조제
나. 맞춤형 고구마 전분원료 효율적 생산 방법 개발
(1) 고구마 전분의 환형아밀로오스 생산을 위한 적합품종 및 재배법 확립
(2) 기능성 탄수화물 생산용 고구마원료 대량 증식
다. 고구마 전분을 이용한 기능성 탄수화물 효율적 생산 방법 개발
(1) 기능성 탄수화물 효율적 생산방법 개발
(2) 기능성 탄수화물 특성 검정
3. 제1협동과제 : 전분활성효소를 이용한 기능성 탄수화물 신소재 제조 및 특성
가. 전분활성 효소를 이용한 환형아밀로스 제조 및 분리 최적화
(1) 전분활성 효소인 TAαGTase 분리 및 정제
(2) cycloamylose 제조 조건 최적화
(3) cycloamylose 이화학적 특성 규명
- HPAEC, MALDI-TOF MS, Iodine absorption spectra
나. 환형 아밀로스 혼합물의 중합도별 분리 및 복합체 형성능 규명
(1) 환형아밀로스 혼합물의 중합도별 분리 최적화 및 분리여부 확인
- HPA EC, MALDI-TOF MS
(2) 모델코어물질과 중합도별 분리한 환형아밀로스와의 복합체 형성 분리능 규명
- ITC, NMR
다. 선별된 기능성 코어물질과 환형아밀로스의 포접 복합능 규명
(1) 환형아밀로스와 lysolecithin (LL)과의 interaction : ITC, NMR
(2) 환형아밀로스와 vitamin K3 (VK3)와의 interaction : ITC, NMR

Abstract ▼

Sub-project 1 : Evaluation of rice varieties for modified starch through carbohydrate active enzymes
Starch is the major component of rice grains, which are widely recognized as an important source of this carbohydrate in the food industry. Of note in particular, the ratio of amylose and amylopectin has been revealed to be an important determinant of the cooking and eating quality of milled rice as well as the outcomes of processing. In our current study we focused on elucidating the structures and physicochemical characteristics of starch contents of rice cultivars with differing amylose levels with the aim of obtaining useful basic data that will assist in the selection of starch materials and thereby the selection of new functional rice varieties.
Significant differences in the amylose content and maximum absorption wavelength (λmax) were observed among the rice cultivars. The starches from these rice cultivars were thereby found to exhibit significantly different amyloectin chain length distributions. The proportion of short chains in the Goami cultivar, which has a high amylose content, was higher than in any of the other cultivars, whereas the proportion of long chains with a DP ≥ 37 was the lowest in this strain. These X-ray diffraction patterns were A-type in all of strains except Goami 2 and Goami 4, with the main diffraction peaks observed at 15, 17, 18 and 23 degrees 2θ. The Goami 2 variety displayed B-type pattern however, which is characterized by tube and root starches. B-type starches have higher proportions of longer amylopectin chains than their A-type counterparts. We could also found that the rice cultivars had different levels of the enzymatic hydrolysis rate and TAaGTase-modified starches. These findings suggest that the composition and chemical structure of the starch content is a major determinant of both the gelatinization and functional properties of rice.

목차 Contents

표지 ... 1
제출문 ... 2
요약문 ... 4
SUMMARY ... 17
제1장 서 론 ... 23
제1절 연구개발의 목적 ... 23
제2절 연구개발의 필요성 ... 24
제3절 연구개발의 범위 ... 26
제2장 국내외 기술개발 현황 ... 30
제1절 국내연구동향 ... 30
제2절 국외연구동향 ... 32
제3절 국내외 연구현황 비교 및 필요 연구 분야 ... 34
제3장 연구개발수행 내용 및 결과 ... 36
제1절 쌀 전분의 효소적 맞춤형 구조변형에 적합한 유전자원 탐색 및 품종 선발 ... 36
제2절 전분활성효소를 이용한 고구마의 기능성 탄수화물 신소재 개발 ... 49
제3절 전분활성효소를 이용한 기능성 탄수화물 신소재 제조 및 특성 ... 58
제4절 기능성 탄수화물 신소재 개발용 전분활성 효소의 생산 및 이용 ... 104
제5절 기능성 탄수화물 응용기술 개발 ... 115
제6절 연속적인 선택적 카르복실화 반응에 의한 수용성과 기능성이 향상된 변형 다당류의 생산 및 식이섬유 음료 신제품에의 적용 ... 131
제4장 연구개발목표 달성도 및 대외기여도 ... 180
제1절 목표대비 달성도 ... 180
제2절 정량적 성과 ... 184
제5장 연구개발결과의 활용계획 ... 187
제6장 중요 변동사항 ... 188
제7장 참고문헌 ... 189
전분활성효소를 이용한 기능성 탄수화물 소재 적합 쌀 전분 특성 ... 196
전분활성효소를 이용한 고구마 환형아밀로오스 제조 ... 197
전분활성효소를 이용한 기능성 탄수화물 신소재 개발 ... 198
전분활성효소를 이용한 기능성 탄수화물 신소재 개발 ... 200
기능성 탄수화물 신소재 응용기술 개발 ... 202
연속적인 선택적 카르복실화 반응에 의한 수용성과 기능성이 향상된 변형다당류의 생산 및 식이섬유 음료 신제품에의 적용 ... 203

표/그림 (170)

표 Amylose contents and λmax values of iodine absorption of the indicated rice starches
표 Amylopectin chain length distributions in different rice starches.
표 SEM micrographs of isolated starch granules of rice variety
표 Amylose contents and wavelength in λmax of iodine absorption of starches of each endosperm layer in rice grain
표 Amylopectin chain length distribution of each endosperm layer in rice grain
표 Distribution of amylose content of Korean germplasm rice
표 Variation of amylopectin chain length distribution of 103 Korean germplasm
표 Variation of amylopectin chain length of 103 Korean germplasm
표 The yield of cycloamylose of various rice starch were treated with 100U/g of TAα GTase for 6hrs.
표 MALDI-TOF chromatogram of molecular mass distribution of cycloamylose from various rice starch treated with TAαGTase (100U/g) for 6hrs.
표 Amylopectin chain length distributions in different rice starches
$X-ray diffraction patterns of starches form rice cultivars with diverse amylose contents.$ 표 X-ray diffraction patterns of starches form rice cultivars with diverse amylose contents.
표 Hydrolysis rate of different rice starches using glucoamylase (Left) and α-amylase (Right).
표 Gelatinization and endotherm characteristics of starches from rice cultivars
표 Pasting properties of flours and starches from rice cultivars
표 고구마 품종별 전분 호화특성
표 고구마 품종 및 재배기간에 따른 전분의 아밀로오스 함량
표 고구마 품종별 전분의 주사전자현미경(SEM) 사진
표 재배기간에 따른 고구마 품종별 X-ray 굴절률 변화
표 시차주사열량계를 이용한 재배기간에 따른 호화온도변이
표 고구마 품종별 재배기간에 따른 아밀로펙틴의 중합도 측정
표 고구마 품종별 아밀로펙틴 중합도(재배기간 120일)
표 TAαGTase 100U/g으로 6시간 동안 처리한 고구마 전분의 CA 수율
표 120일 재배 고구마 품종별 전분 특성
표 TAαGTase(100U/g)로 6시간 동안 처리한 고구마 전분으로부터 얻은 CA의 분자량 분포에 대한 MALDI-TOF chromatogram,
표 A schematic diagram of the procedure for producing cycloamylose using TAαGTase.
표 HPAEC analysis of cycloamyloses produced by TA αGTase
표 HPAEC analysis of cycloamyloses produced by TAαGTase.
표 HPAEC analysis for amylose and cycloamyloses;
표 MALDI-TOF MS analysis of cycloamylose.
표 Theoretical molecular masses of noncyclic and cyclic glucans, and molecular masses of glucans modified by TAαGTase determined by TOF-MS analysis
표 Phase-solubility diagrams of (a) quercetin/β-cyclodextrin complexes and (b) quercetin/cycloamylose complex in water.
표 Phase-solubility diagrams of (a) quercetin/β-cyclodextrin complexes and (b) quercetin/cycloamylose complex in water.
표 Phase-solubility diagram of (a) quercetin/β-cyclodextrin complexes and (b) quercetin/cycloamylose complex in water (n=3).
표 Aqueous solution of (a) quercetin/β-CD complex, (b) quercetin/CA complex, and (c) quercetin/12 mM CA complex.
표 Apparent stability constant (Kc) of quercetin/β-CD or CA inclusion complexes
$Powder X-ray Diffractogram of Quercetin (a); β-CD (b); Quercetin/β-CD sample mixture (c); Quercetin/β-CD physical mixture (d); Quercetin/β-CD freeze-dried product (e).$ 표 Powder X-ray Diffractogram of Quercetin (a); β-CD (b); Quercetin/β-CD sample mixture (c); Quercetin/β-CD physical mixture (d); Quercetin/β-CD freeze-dried product (e).
$Powder X-ray Diffractogram of Quercetin (a); CA (b); Quercetin/CA sample mixture (c); Quercetin/CA physical mixture (d); Quercetin/CA freeze-dried product (e).$ 표 Powder X-ray Diffractogram of Quercetin (a); CA (b); Quercetin/CA sample mixture (c); Quercetin/CA physical mixture (d); Quercetin/CA freeze-dried product (e).
표 Differential scanning calorimetry thermograms of Quercetin (a); β-CD (b); Quercetin/β-CD simple mixture (c); Quercetin/β-CD physical mixture (d); Quercetin/β-CD freeze-dried product (e).
표 Differential scanning calorimetry thermograms of Quercetin (a); CA (b); Quercetin/CA sample mixture (c); Quercetin/CA physical mixture (d); Quercetin/CA freeze-dried product (e).
표 Chromatograms of cycloamylose. Chromatographic conditions
표 Chromatograms of cycloamylose.
표 Chromatograms of cycloamylose.
표 Separation of CM on a Sunfire C18 column (10μm, 30×150mm).
표 The yield of each cyclic α-(1→4) glucan from cycloamylose
표 H-NMR chemical shifts (ppm) of cyclic glucans in D2O at 298.2K
표 HPAEC elution profile of each cyclic α-(1→4)-glucan isolated from prep-HPLC.
표 MALDI-TOF-MS analysis of each cyclic α-(1→4)-glucan isolated from prep-HPLC.
표 (a) Experimental data for titrations of 8㎕ aliquots of 1.88 mM SDS into a reaction cell containing 0.05 mM CMs, CA25, CA30, CA35 and CA40 at 25℃.
표 DP dependences of (a) stoichiometry (n), (b) binding constants (Ka)at25.0℃ for CA23-CA45.
표 DP dependences of ∆H (▲), -T∆S (■) and ∆G (◇) at 25.0℃ for CA23-CA45.
표 1H NMR spectra of (a) SDS, (b) cyclic glucans alone, and (c) cyclic glucans with SDS solution.
표 H NMR spectra of (a) SDS; shown Fig 16, (b) isolated CA molecules alone, and (c) CA23-CA45 and SDS mixture.
표 Proton of the 600 MHz ROESY spectrum of the CA26-SDS mixture in D2O at 298.2K.
표 The molecular structures of (a) SDS and (b) a part of CA26 molecules; and numbered of each atom. (c) ROESY cross-peak, expanded and amplified from Fig. 12, of CA26-SDS mixture.
표 Thermodynamic parameters for CA-guest complexes at T=298K
표 (a) Experimental data for titrations of 8㎕ aliquots of 1.88 mM LL and V3 into a reaction cell containing 0.05 mM CA at 25℃.
표 Changes in free energy, enthalpy, and entropy for inclusion complexation of CA with LL, VK3, and SDS.
표 The molecular structures of (a) LL and (b) VK3 molecules; and numbered of each atom.
표 H chemical shifts corresponding to cycloamylose in the presence and absence of LL and VK3 [△δ = δ(complex) - δ(free)]
표 A portion of ROESY cross-peak of CA-LL.
표 Phase solubility diagrams of VD3-CA or VD3-CD system in water at 40℃: (a) β-CD, (b) CA, (c) α-CD, (d) γ-CD.
표 FT-IR spectra of (a) VD3, (b) CA, (c) PM, (d) KM, (e) CPM
$Powder X-ray Diffractogram of (a) VD3, (b) CA, (c) PM, (d) KM, (e) CPM.$ 표 Powder X-ray Diffractogram of (a) VD3, (b) CA, (c) PM, (d) KM, (e) CPM.
표 Differential scanning calorimetry thermograms of (a) VD3, (b) CA, (c) PM, (d) KM, (e) CPM.
표 H NMR spectrum of VD3-CA complex in DMSO-d6 at 600 MHz at 297K.
표 The chemical shifts of the protons belonging to VD3, CA and to the inclusion complex
표 A portion of 600 MHz 2D ROESY symmetrized spectrum of VD3-CA complex prepared by CPM.
표 pR2CGTI-5의 구성 (a) 및 CGTase I-5 의 정제 (b)
표 p6XHTKTSαGT의 구성 및 TSαGTase 의 정제
표 HPAEC를 이용한 환형말토덱스트린 분석 (beta amylase 처리 후 결과).
표 MALDI-TOF MS를 이용한 환형말토덱스트린 구조 분석.
표 본 연구에서 사용된 환형아밀로오스의 제조방법.
표 그림 1.
표 그림 2.
표 그림 3.
표 그림 4.
표 그림 5.
표 그림 1.
표 그림 2.
표 그림 3.
표 그림 4.
표 Structure of citral.
표 Structure of β-carotene.
표 Structure of curcumin.
표 Structure of quercetin.
표 Structure of flurbiprofen.
표 Structure of valsartan.
표 Structure of silybin.
표 HPLC raw data.
표 Calibration curve for assay of flurbiprofen.
표 HPLC raw data.
표 Calibration curve for assay of varsartan.
표 HPLC raw data.
표 Calibration curve for assay of Silymarin.
표 Aqueous solubility of poorly water-soluble materials
표 Scanning electron micrographs: (a) flurbiprofen powder (1000x); (b) solid dispersion (800x).
표 Scanning electron micrographs (1000): (a) ibuprofen (b) cycloamylose (c) solid dispersion.
표 Differential scanning calorimetric thermograms:
$X-ray powder diffraction:$ 표 X-ray powder diffraction:
표 Dissolution profile of the flurbiprofen from the solid dispersion and commercial product.
표 Dissolution profile of the ibuprofen (A) and dissolution kinetics (B) of the drug from the solid dispersion and ibuprofen powder.
표 Plasma concentration–time profiles of the drug after oral administration of the solid dispersion and commercial product to rats.
표 Plasma concentration–time profiles of the drug after oral administration of the solid dispersion and ibuprofen powder to rats.
표 Pharmacokinetic parameters
표 Scheme of selective oxidation mediated by sodium bromide-free TEMPO/sodium hypochlorite system.
표 Schematic diagram of reactor system for the selective oxidation.
표 Photographic representation of rice (a cross section)
표 Nutrient composition of rice grain, rice bran, and sweet potato flour
표 Levels of TEMPO, sodium hypochlorite, pH of reactant, and reaction temperature for selective oxidation
표 Time course of selective oxidation mediated by TEMPO/NaOCl.
표 Effect of reaction conditions on consumption of 0.5 N NaOH during selective oxidation of rice grain.
표 Effect of reaction conditions on consumption of 0.5 N NaOH during selective oxidation of rice bran.
표 Effect of reaction conditions on consumption of 0.5 N NaOH during selective oxidation of sweet potato powder.
표 Effect of reaction conditions on consumption of 0.5 N NaOH during selective oxidation of sweet potato residue.
표 Relative DO (%) of rice grain, rice bran, SP powder, and SP residue according to reaction conditions
표 Effect of reaction conditions on initial velocity of selective oxidation of rice grain.
표 Effect of reaction conditions on initial velocity of selective oxidation of rice bran.
표 Effect of reaction conditions on initial velocity of selective oxidation of sweet potato powder.
표 Effect of reaction conditions on initial velocity of selective oxidation of sweet potato residue.
표 Initial velocity of selective oxidation of rice bran, SP powder, and SP residue according to reaction conditions
표 Independent variables and their levels for RSM
표 Central composite experimental design consisting of the 3-factors and 5-levels
표 Schematic procedure for the semi-continuous process of selective oxidation.
표 Response surface plots showing the effects of variables on the DO (%) of rice bran:
표 Response surface plots showing the effects of variables on the DO (%) of sweet potato residue:
표 Contour plots of the DO (%) of rice bran: (a) TEMPO and NaOCl; (b) NaOCl and pH; (c) TEMPO and pH.
표 Contour plots of the DO (%) of sweet potato residue: (a) TEMPO and NaOCl; (b) NaOCl and pH; (c) TEMPO and pH.
표 Regression coefficients of the predicted polynomial model (Rice bran)
표 Regression coefficients of the predicted polynomial model (SPR)
표 Analysis of variance (ANOVA) of independent variables for the second-order response surface model (Rice bran)
표 Analysis of variance (ANOVA) of independent variables for the second-order response surface model (sweet potato residue)
표 Optimum conditions for the selective oxidation of rice bran by response surface methodology.
표 Optimum conditions for the selective oxidation of sweet potato residue by response surface methodology.
표 Optimum conditions for the TEMPO-mediated selective oxidation by respond surface methodology (RSM)
표 Relative DO (%) of water-insoluble polysaccharides by TEMPO-mediated oxidation under the optimum conditions
표 13C-NMR spectra of native (a) and oxidized (b) rice bran.
표 13C-NMR spectra of native (a) and oxidized (b) sweet potato residue (SPR).
표 IR spectra of native (a) and oxidized (b) rice bran
표 IR spectra of native (a) and oxidized (b) sweet potato residue (SPR).
표 Time course of selective oxidation mediated by TEMPO/sodium hypochlorite in semi-continuous system.
표 Changes in reaction time of the TEMPO-mediated selective oxidation according to recycling number in semi-continuous system.
표 Cookie, bread and scone, 486; normal products, 327; products containing oxidized rice bran
표 Soft drink and orange juice, 486; normal products, 327; products containing oxidized rice bran
표 Ingredients and chemical conditions used to produce soft drink containing oxidized rice bran
표 Ingredients and chemical conditions used to produce orange juice containing oxidized rice bran
표 The mixing process of soft drink
표 The manufacturing process of soft drink
표 The mixing process of orange juice
표 The manufacturing process of orange juice
표 Overall acceptance analysis of cookie containing oxidized rice bran and normal product
표 Acceptance, flabor, texture and aftertaste analysis of cookie containing oxidized rice bran and normal product
표 Acceptance, flabor, texture and aftertaste analysis of cookie containing oxidized rice bran and normal product
표 Properties of recognition strength between cookie containing oxidized rice bran and normal product.
표 Overall acceptance analysis of bread containing oxidized rice bran and normal product
표 Acceptance, flabor, texture and aftertaste analysis of bread containing oxidized rice bran and normal product
표 Acceptance, flabor, texture and aftertaste analysis of bread containing oxidized rice bran and normal product
표 Properties of recognition strength between bread containing oxidized rice bran and normal product.
표 Overall acceptance analysis of scone containing oxidized rice bran and normal product
표 Acceptance, flavor, texture and aftertaste analysis of scone containing oxidized rice bran and normal product
표 Acceptance, flavor, texture and aftertaste analysis of scone containing oxidized rice bran and normal product
표 Properties of recognition strength between scone containing oxidized rice bran and normal product.
표 Overall acceptance analysis of soft drink containing oxidized rice bran and normal product
표 Acceptance, flabor, texture and aftertaste analysis of soft drink containing oxidized rice bran and normal product
표 Acceptance, flabor, texture and aftertaste analysis of soft drink containing oxidized rice bran and normal product
표 Properties of recognition strength between soft drink containing oxidized rice bran and normal product.
표 Overall acceptance analysis of orange juice containing oxidized rice bran and normal product
표 Acceptance, flavor, texture and aftertaste analysis of orange juice containing oxidized rice bran and normal product
표 Acceptance, flavor, texture and aftertaste analysis of orange juice containing oxidized rice bran and normal product
표 Properties of recognition strength between orange juice containing oxidized rice bran and normal product.

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내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 제목(한글), 저자명(한글), 발행일자, 전자원문, 초록(한글), 초록(영문) 관리번호, 제목(한글), 제목(영문), 저자명(한글), 저자명(영문), 주관연구기관(한글), 주관연구기관(영문), 발행일자, 총페이지수, 주관부처명, 과제시작일, 보고서번호, 과제종료일, 주제분류, 키워드(한글), 전자원문, 키워드(영문), 입수제어번호, 초록(한글), 초록(영문), 목차
저장형식	Text(ASCII format) Excel format
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증

전분 활성 효소를 이용한 기능성 탄수화물 신소재 개발
Developing of functional carbohydrates using carbohydrate active enzymes 원문보기