Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Physicochemical Characteristics of Galactomannan by Fractionation to Evaluate Heterogeneity

불균일 성질을 평가하기 위한 분획화된 galactomannan의 이화학적 특성

  • Received : 2013.04.11
  • Accepted : 2013.05.09
  • Published : 2013.08.31
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Abstract

Heterogenous samples of locust bean gum (galactomannan) were prepared into homogeneous substances. Locust bean gum was fractioned using ammonium sulfate (14.11-23.08%, w/w). The intrinsic viscosity was obtained by extrapolating reduced viscosity versus concentration by using an Ubbelohde viscometer. The ranges of intrinsic viscosity for fractions that not included protein (F3-F6) and fractions that included protein (F1-F2) were 9.89-8.10 and 8.44-4.59, respectively. Values for Huggins' coefficient (k'), which depends on physical interactions, were 0.46-0.78. Increasing ammonium sulfate concentration was associated with a weak trend towards lower molecular weight and intrinsic viscosity by size-exclusion chromatography (SEC): $M_w$ ranged from 674 to 617 kg/mol and [${\eta}$] from 9.80 to 8.10 dL/g between F3 and F6. The evaluations of those fractions by using SEC and the Ubbelohde viscometer produced very similar values, as predicted. We verified the application of a gradient of ammonium sulfate to precipitate locust bean gum into fractions of different molecular size and show structural variations.

Mannose 기본구조에 galactose 치환체 비율이 4:1인 화학적 구조와 평균 분자량이 1,050 kg/mol인 LBG를 황산암모늄 침전법을 사용하여 분자량 크기별로 분획하여 분획물질들 간에 서로 다른 물리화학적 성질을 확인하였다. 황산암모늄을 맑은 용액에 천천히 첨가한 후 얻어진 침전물을 원심분리에 의해 모집, 탈이온수에 대한 투석 후 동결상태에서 건조시켜서 첫 번째 분획물을 얻었고 동일한 방법으로 각 단계별 분획물을 여섯 단계 까지 얻었다(F1-F6). 수득율은 F1 7.1% 부터 F6 6.35%까지 총 65%였다. 각 분획물들의 묽은 농도범위 0.05 g/dL 이하에서 Ubbelohde viscometer로 흐르는 시간을 측정하여 상대점성도, 비점성도, 유도점성도 및 본성점성도를 구하였으며 그 값은 F1:8.44, F2:4.59, F3:9.89, F4:8.80, F5:8.30, F6:8.10 dL/g이었다. F1과 F2 분획물이 갖는 본성 점성도 값은 기대치보다 작았고 그 원인은 단백질 성분을 포함하는 것으로 판단되어 함량을 측정한 결과 전체 단백질 함량 3.45%중에 F1, F2가 2.59%를 포함하고 있음을 확인하였다. 또한 고분자 물질의 용질-용매간의 상호작용과 응집상태에 의존하는 상태를 나타내는 k' (Hugg. Coeff.)값을 측정한 결과 값의 범위는 0.463-0.781이였고 점성도 값이 클수록 k'값이 커지는 경향을 나타내었다. 분획물들의 체류용량에 대한 RI detector 크로마토그램은 4개의 분획물들의 다당류는 6.0에서 9.0 mL에서 용리되었고 염 혹은 oligomeric sugar 성분은 10에서 11 mL에서 용리되었음을 나타내었다. 중량 평균 분자량, 수 평균 분자량, 회전 반경 및 본성 점성도 값들은 $c_p=0.5mg/mL$, dn/dc = 0.145의 RI 피크면적에 기초를 둔 OmniSEC 프로그램으로 계산되었다. 본성 점성도 값은 F3: 10.15, F4: 9.99, F5: 9.35, F6: 9.31이었고 $M_w$값 범위는 617-674 kg/mol, $M_n$값 범위는 324-423%, recovery 72.44-101.61%였다. Ubbelohde viscometer와 SEC로 측정한 각 분획물들의 본성 점성도 값을 비교한 결과, 측정에 사용한 기기 원리와 계산식의 영향으로 수치에는 미소한 차이가 있었으나 분획물 순서대로 본성점성도가 감소하는 경향은 동일하여 황산암모늄 침전법에 의한 LBG 분획화가 차별화 있게 잘 이루어졌음을 확인하였다. 화학물질에 대한 안정제 및 식품첨가제로 주로 사용되고 있는 LBG 복합물질을 단순물질로 분획화하여 균일한 성분의 물리화학적 성질을 연구한 것은 안정제 및 첨가제의 효과를 높이고 사용범위가 확대될 수 있을 것으로 기대된다.

Keywords

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