학술논문
개암버섯 균사체의 영양성분 분석 및 생리활성 측정 / Evaluation of Nutritional Value and Biological Activities of Naematoloma sublateritium Mycelium
Document Type
Dissertation/ Thesis
Source
Subject
Language
English
Abstract
야생에서 채집한 개암버섯 (Naematoloma sublateritium) 균사체를 5.8S ribosomal DNA 염기서열분석을 통하여 동정하였다. 동결건조된 개암버섯 균사체의 일반성분, 수용성비타민, 유기산, 유리당, 유리아미노산 조성을 분석하였다. 일반성분은 탄수화물 57.97%, 조단백 29.35%, 회분 4.81%, 조지방 1.82%, 수분 6.04%로 분석되었다. 개암버섯 균사체의 수용성비타민은 비타민 B3 (15.79 mg/g), 비타민 C (7.35 mg/g), 비타민 B6 (1.89 mg/g), 비타민 B9 (59.05 μg/g), 비타민 B1 (27.16 μg/g), 비타민 B2 (18.52 μg/g) 6종이 분석되었다. Fumaric acid, malic acid, pyroglutamic acid, sebacic acid, succinic acid 등 5종 유기산이 검출되었고 galactose (10.99 mg/g), trehalose (6.30 mg/g) 및 sorbitol (11.47 mg/g)이 주요 유리당 및 당알콜인 것으로 확인되었다. 이외에도 L-serine, L-threonine, L-histidine 등이 개암버섯 균사체에 함유된 주요 유리아미노산인 것으로 나타났다.항산화활성을 측정하기 위하여 개암버섯 균사체 메탄올 추출물을 제조하여 이 추출물에서 n-hexane, chloroform, ethyl acetate, n-butanol, water 분획을 분리하였다. Chloroform 분획은 아질산염소거능을 제외한 대부분의 평가에서 가장 높은 항산화활성을 나타내었다. Ferric reducing antioxidant power는 chloroform 분획에서 299.24 μmol FeSO4•7H2O/g으로 나타났고, ABTS 라디칼 소거능은 649.40 μmol/g 이었다. 이 분획은 406.59 μg/mL의 농도에서 DPPH 라디칼을 50% 소거하였다. 유리 페놀산을 ESI LC-MS/MS 분석을 통해 동정한 결과, 4-hydroxybenzoic acid, α-resorcylic acid, 4-coumaric acid, gentisic acid, salicylic acid 등 5종이 검출되었다.Chloroform 분획이 LPS로 염증이 유도된 대식세포에 미치는 영향을 조사한 결과, iNOS, p-STAT1, p-NFκB, p-ERK, p-JNK의 발현을 현저하게 저해하였지만 COX-2와 p-p38 단백질의 발현은 저해하지 않았다. Chloroform 분획은 LPS로 염증이 유도된 대식세포에서 NFκB, STAT1, ERK, JNK 등의 신호전달을 하향 조절하여 iNOS의 발현을 저해하는 것으로 나타났다. 본 연구에서는 개암버섯 추출물에 대한 항산화활성 및 항염증활성을 조사함으로서 개암버섯의 산업응용 가능성을 제시하였다.
A wild mushroom strain was isolated from rotten wood and identified as Naematoloma sublateritium species by 5.8S ribosomal DNA sequence analysis. The freeze-dried mycelium of N. sublateritium was mainly composed with carbohydrate (57.98%) followed by crude protein (29.35%), ash (4.81%), crude fat (1.82%) and moisture (6.04%). Thiamin (27.16 μg/g), riboflavin (18.52 μg/g), niacin (15.79 mg/g), pyridoxine (1.89 mg/g), folic acid (59.05 μg/g), and vitamin C (7.35 mg/g) were identified as major vitamins and fumaric acid, malic acid, pyroglutamic acid, sebacic acid, and succinic acid were quantified in organic acid analysis. Galactose (10.99 mg/g), trehalose (6.30 mg/g) and sorbitol (11.47 mg/g) were found to be major sugars and sugar alcohol. L-serine, L-threonine, and L-histidine were main free amino acids among sixteen analyzed free amino acids. Various subfractions were obtained from methanol extract of N. sublateritium mycelium by systemic extraction with n-hexane, chloroform, ethyl acetate, n-butanol and water. The chloroform subfraction exhibited the strongest antioxidant activity except nitrite scavenging activity. The trolox equivalent antioxidant capacity (TEAC) of chloroform subfraction was 649.40 μmol/g of sample. The same fraction also scavenged DPPH radical by 50% at 406.59 μg/mL. The highest ferric reducing antioxidant power was found in the same subfraction (299.24 μmol FeSO4•7H2O /g), and chloroform and ethyl acetate fractions exhibited the highest contents of total phenolic compounds, with ferulic acid equivalents of 31.66 and 48.57 mg/g, respectively. The level of flavonoid was at maximum in chloroform subfraction (18.66 mg/g) and the concentration of phenolic compounds may be correlated with the antioxidant activities of the subfractions. 4-Hydroxybenzoic acid was detected as the highest level of phenolic acid in a free phenolic form. Five phenolic acids such as 4-hydroxybenzoic acid, α-resorcylic acid, 4-coumaric acid, gentisic acid, and salicylic acid were found as the major free phenolics by using LC-MS/MS. The chloroform subfraction decreased the nitric oxide production, and it inhibited the expression of iNOS, p-STAT1, p-NFκB, p-ERK, and p-JNK in LPS-induced RAW 264.7 mouse macrophages. The inhibition effect could not be observed on COX-2 and p-p38. The chloroform extract suppressed LPS-induced inflammatory signaling through regulating the NFκB, STAT1, ERK and JNK, which leads to inhibition of iNOS expression. N. sublateritium extract seems to prevent the oxidative and inflammatory signaling in vitro and it increase the possibility of industrial application.
A wild mushroom strain was isolated from rotten wood and identified as Naematoloma sublateritium species by 5.8S ribosomal DNA sequence analysis. The freeze-dried mycelium of N. sublateritium was mainly composed with carbohydrate (57.98%) followed by crude protein (29.35%), ash (4.81%), crude fat (1.82%) and moisture (6.04%). Thiamin (27.16 μg/g), riboflavin (18.52 μg/g), niacin (15.79 mg/g), pyridoxine (1.89 mg/g), folic acid (59.05 μg/g), and vitamin C (7.35 mg/g) were identified as major vitamins and fumaric acid, malic acid, pyroglutamic acid, sebacic acid, and succinic acid were quantified in organic acid analysis. Galactose (10.99 mg/g), trehalose (6.30 mg/g) and sorbitol (11.47 mg/g) were found to be major sugars and sugar alcohol. L-serine, L-threonine, and L-histidine were main free amino acids among sixteen analyzed free amino acids. Various subfractions were obtained from methanol extract of N. sublateritium mycelium by systemic extraction with n-hexane, chloroform, ethyl acetate, n-butanol and water. The chloroform subfraction exhibited the strongest antioxidant activity except nitrite scavenging activity. The trolox equivalent antioxidant capacity (TEAC) of chloroform subfraction was 649.40 μmol/g of sample. The same fraction also scavenged DPPH radical by 50% at 406.59 μg/mL. The highest ferric reducing antioxidant power was found in the same subfraction (299.24 μmol FeSO4•7H2O /g), and chloroform and ethyl acetate fractions exhibited the highest contents of total phenolic compounds, with ferulic acid equivalents of 31.66 and 48.57 mg/g, respectively. The level of flavonoid was at maximum in chloroform subfraction (18.66 mg/g) and the concentration of phenolic compounds may be correlated with the antioxidant activities of the subfractions. 4-Hydroxybenzoic acid was detected as the highest level of phenolic acid in a free phenolic form. Five phenolic acids such as 4-hydroxybenzoic acid, α-resorcylic acid, 4-coumaric acid, gentisic acid, and salicylic acid were found as the major free phenolics by using LC-MS/MS. The chloroform subfraction decreased the nitric oxide production, and it inhibited the expression of iNOS, p-STAT1, p-NFκB, p-ERK, and p-JNK in LPS-induced RAW 264.7 mouse macrophages. The inhibition effect could not be observed on COX-2 and p-p38. The chloroform extract suppressed LPS-induced inflammatory signaling through regulating the NFκB, STAT1, ERK and JNK, which leads to inhibition of iNOS expression. N. sublateritium extract seems to prevent the oxidative and inflammatory signaling in vitro and it increase the possibility of industrial application.