Soybean Oil 및 Flaxseed Oil 첨가 배양시 탄수화물 첨가수준에 의한 반추미생물의 Bio-hydrogenation과 CLA 생성에 미치는 효과
Effect of the Level of Carbohydrates on Bio-hydrogenation and CLA Production by Rumen Bacteria When Incubated with Soybean Oil or Flaxseed Oil In vitro
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본 실험은 대두유 또는 아마유를 in vitro 방법으로 배양 할 때, 탄수화물원의 첨가수준이 반추위 박테리아에 의한 bio-hydrogenation과 CLA 생성에 미치는 효과를 조사하였다. 4수준(0%, 0.3%, 0.6% 그리고 0.9%, w/v)의 혼합된 탄수화물원(glucose, cellobiose, soluble starch, 1:1:1, w/w/w)과 두 종류의 oil을 cellulose powder에 흡착시킨 형태로 각각 60mg씩 인공타액(120ml)과 반추위액(30ml)이 혼합된 배양액(150ml)에 넣은 다음 39℃에서 12시간동안 혐기적으로 배양하였다. 배양액의 pH와 암모니아 농도는 두 종류 oil을 첨가한 배양액 모두에서 탄수화물원의 첨가 수준이 높을수록 pH와 암모니아 농도가 낮았다(P<0.05). 탄수화물원의 첨가 수준이 증가할수록 total VFA 생성량 역시 증가되었으나(P<0.01) 첨가한 oil 간의 차이는 없었다. 배양시간이 경과됨에 따라 탄수화물원의 첨가수준이 높을수록 propionate의 조성비율이 증가된 반면(P<0.001) acetate와 butyrate의 조성비율은 감소되었다. 배양 후 3시간이 경과하였을 때 배양액 내 oleic acid의 조성비율은 대두유에 비하여 아마유를 첨가한 배양액에서 낮았으나(P<0.001) linoleic acid의 비율은 높았다(P<0.001). 이와는 달리 탄수화물원의 수준이 증가될수록 stearic acid(P<0.05), CLA(P<0.01) 및 cis-9, trans-11 CLA(P<0.001)의 조성비율은 감소되었으나, linoleic acid의 조성 비율은 증가되었다(P<0.05). Linolenic acid의 조성비율에 있어서는 첨가된 oil의 종류와 첨가된 탄수화물원의 수준간의 상호작용이 있는 것으로 나타났는데(P<0.001), 12시간의 배양종료 후 대두유 첨가구에 비해 아마유 첨가구에서 stearic acid(P<0.01), oleic acid(P<0.001), 그리고 trans-11C18:1(P<0.01)의 조성비율이 감소된 반면, linoleic acid(P<0.001)와 linolenic acid(P<0.01)의 조성비율은 증가되었다. 탄수화물원의 첨가수준이 증가될수록 stearic acid와 총 CLA의 조성비는 감소되었으나(P<0.01), trans-11-C18:1(P<0.05)와 linoleic acid(P<0.01)의 조성비율은 증가되었다. 배양 12시간 후 배양액 내의 oleic acid (P<0.05), linoleic acid(P<0.05) 및 linolenic acid(P<0.01)의 조성비율에 있어서는 첨가한 oil의 종류와 첨가한 탄수화물원의 수준간의 상호작용이 있었는데, 탄수화물원의 첨가수준이 감소됨에 따라 cis-9, trans-11 CLA와 trans-10, cis-12 CLA의 조성비 역시 감소되는 경향이었으나 첨가한 oil의 종류에 대한 영향은 거의 받지 않았다. 본 실험의 결과를 종합해 볼 때, 탄수화물의 첨가수준과 oil의 첨가는 반추미생물의 bio-hydrogenation 작용 및 CLA 생성에 영향할 수 있는 것으로 여겨진다.
Abstract
An in vitro study was conducted to examine the effect of addition level of carbohydrates on fermentation characteristics, and bio-hydrogenation of unsaturated fatty acids by mixed rumen bacteria when incubated with soybean oil or flaxseed oil. Four levels(0%, 0.3%, 0.6% and 0.9%, w/v) of the mixed carbohydrates(glucose, cellobiose, soluble starch, 1:1:1, in weight basis) and oil sources(soybean or flaxseed oil, 60mg in 150ml culture solution) were added to the mixed solution of strained rumen fluid with artificial saliva(1:4, v/v), and incubated anaerobically for 12 hours at 39℃. pH and ammonia-N concentration were lower by increasing the substrate levels at all incubation periods(P<0.05~P<0.001). The propionate proportion increased(P<0.001), but acetic acid and butyric acid decreased(P<0.001) with the substrate level at 6 and 12 h incubations. Oil sources did not influence the proportions of individual VFA. At the end of incubation, the proportions of C18:0(P<0.01), C18:1(P<0.001) and trans-11C-18:1(P<0.01) were reduced but those of C18:2(P<0.001) and C18:3(P<0.01) were enhanced by the addition of flaxseed oil compared to addition of soybean oil. The proportions of C18:0 and total CLA were reduced(P<0.01) but those of trans-11-C18: (P<0.05) and C18:2(P<0.01) were increased with the substrate level when incubated with soybean oil or flaxseed oil. There were interactions(P<0.05) in the proportions of C18:1, C18:2 and C18:3(P<0.01) between oil source and substrate level. The proportions of cis-9, trans-11-CLA and trans-10, cis-12-CLA tended to reduce with substrate level, although there was no significant difference between treatments.
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