소화성 질환의 중요 인자인 Helicobacter pylori를 저해하는 IgY, 목이버섯, 김치와 타락 유산균을 이용하여 생육 저해 상승 효과를 분석하고자 하였다. 동결 건조된 유산균 배양 상징액을 다양한 효소 처리 결과, 지질 분해 효소를 제외하고는 활성을 나타냈다. GC 분석을 통해 유산균 동결건조액의 지방산 조성은 undecanoic acid(C11:0), palmitic acid(C16:0), steraic acid(C18:0), oleic acid(C18:1)가 L. mensenteroides LAB KW5와 S. thermophilus LAB KW15에서 모두 확인되었으며, eicosadienoic acid(C20:2)는 LAB KW5에서만 나타났다. 또한 유산균 동결건조액은 다른 식중독균에서도 spot assay의 결과, 그람 음성균 중에서 특히 E. coli O157:H7, E. coli, C. sakazakii 등에서 생육 저해능이 확인되었다. 목이버섯 추출물은 열수 추출과 ethanol를 이용해 분리하였으며, HPLC를 이용하여 목이버섯 추출 다당체를 분석한 결과, glucuronoxylomannan 혹은 glucomannan이 β-glucan과 함께 존재하는 혼합물일 것으로 되었다. 또한, 면역란은 1차 접종 후 11일째부터 주마다 2회씩 회수하여 IgY를 분리, 정제하였다. 실험을 통해 동결 건조된 유산균 배양 상징액, IgY, 목이버섯 추출액을 혼합 배양하여 배양시간에 따른 생육 저해력의 변화를 확인한 결과, 유산균에 의해 H. pylori의 저해 효과가 있었으며, IgY와 목이버섯의 혼합 시 추가 억제 효과가 있는 것으로 나타났다. 그러므로 동결 건조된 유산균 배양 상징액, IgY, 목이버섯 추출액의 복합처리는 H. pylori를 제어하는데 효과적인 것으로 보인다.

본 연구는 안전하면서 효율적으로 비만을 개선시키는데 도움이 될 수 있는 식품소재로 목이버섯을 선정하여 복부비만이 있는 중년 여성의 골밀도 및 혈청지질농도에 미치는 영향에 대하여 알아보고자 하였다. 30～50대의 복부비만 여성 30명에게 4주 동안 대조군은 목이버섯을 섭취시키지 않았고, 실험군은 목이버섯을 식사와 함께 섭취하도록 하였다. 실험 전 후 골밀도(T-score, Z-score), 총 콜레스테롤, HDL-콜레스테롤, LDL-콜레스테롤, 중성지방을 측정하였다. 연구결과, 목이버섯 섭취군과 대조군 간에 T-score와 Z-score는 유의한 차이를 보이지 않았지만, 목이버섯 섭취군의 혈청 총 콜레스테롤, LDL-콜레스테롤 및 중성지방 수준이 대조군에 비해 유의하게 소되었다.

목이버섯(Auricularia)의 이름은 그리스어인 “Auricula"에서 왔으며 ”귀(ear)"라는 뜻이다. 따라서 보통 나무귀(tree ear), Jew's 귀 또는 귀버섯이라고 불린다. 즉 자실체의 형태가 귀 와 비슷하고, 촉감이 고무질과 젤라틴질에 의해 귀처럼 느껴지기 때문이다. Lowy에 의해 분류된 목이버섯 10여 종류 가운데 목이(A. auricula)와 털목이(A. polytricha)가 가장 인기있는 버섯으로 두종류 모두 사물기생균이다. 목이는 최초로 인공재배된 버섯으로 보고되었는데, A.D. 600년경부터 중국에서 재배되어왔으며, 최근 국내에서 수요가 증가하고있는 버섯이다. 시험에 사용한 배지재료로 사용한 참나무톱밥, 미강의 이화학적 특성을 조사한 결과, pH는 참나무톱밥 6.0, 미강 6.4 였으며, T-C는 참나무톱밥 46.6%, 미강 52.4%로 나타났고, T-N는 참나무톱밥이 0.28%, 미강 0.94%로 나타났다. CaO의 경우 참나무톱밥 0.92%으로 나타났으며, 미강 0.07%로 톱밥보다 미강이 낮았다. 목이버섯의 균사생장 최적온도를 구명하고자 15℃에서 35℃까지 5℃ 간격으로 처리하여 균사를 배양한 결과 Fig. 1에서와 같이 25～30℃가 배양적온으로 나타났다. 최적 pH는 전범위에서 비슷하게 생육하였으며, 최적배지는 MEA, 탄소원은 Mannose, 비타민은 Biotin, Pyridoxine, 최적 C/N비는 10:1~1:1로 나타났다. 자실체 발생시험에서 배양소요일수는 31일, 초발이소요일수 15일, 자실체 생육일수 18일로 나타났다. 수량의 경우 배지당 생체중이 295g, 건물중이 31g으로 나타났다. 목이버섯균의 cellulolytic activity 검정결과 congo red 염색에서 pH 7.0, 25℃에서 우수한 활성을 나타내었다. 목이버섯 자실체의 열수추출물에 대하여 SRB법과 MTT법으로 종양세포주(Sarcoma 180, P 388)를 이용하여 항암활성을 비교ㆍ평가하였다. 종양세포주는 7.5, 15, 30㎍/㎖ 그리고 Doxorubicin(DOX, 0.001-10mM)으로 처리하였다. 그 결과 DOX, 목이버섯추출물 종양세포 주에 대하여 농도 의존적으로 억제하는 결과를 보였다. 결론적으로, 이 연구에서 사용된 목이버섯추출물은 Sarcoma 180과 P 388에 항암활성을 보여주었다.

The aim of this study was to investigate the suppressive activity of 70% ethanol extract and its dichloromethane fraction from Auricularia auricula-judae against adipogenesis and lipogenesis in 3T3-L1 preadipocytes. The ethanol extract and its dichloromethane fraction suppressed the differentiation and decreased lipid droplets in vitro. The dose-dependent increasing concentration of glycerol and lower triglycerides accumulation were found significantly (P < 0.05) with the treatment of both fractions from 70% ethanolic Auricularia auricula-judae extract and the glycerol-3-3phosphate dehydrogenase (GPDH) activity was also inhibited by both extracts. Further, the expression of adipogenic mRNAs were investigated by RT-PCR amplification. The key transcriptional factors, PPARγ and C/EBPα were decreased significantly at dose-dependent manner by both extracts of Auricularia auricula-judae. The expressions of LPL and FAS were also decreased by presence of these extracts. The decreased expressions of C/EBPβ, C/EBPγ and ACC1 were observed only by ethanol extract at 300 μg/ml concentration, while the expression of SREBP-1c, GLUT4 and aP2 were not altered in the 3T3-L1 adipocytes. These changes were occurred without the cytotoxic effect of both extracts against 3T3-L1 adipocytes in vitro. A positive control, fenofibrate inhibited the differentiation, triglycerides accumulation through PPARγ signaling by 2/3 reduction of PPARγ expression. Thus, these findings suggest that both extracts of Auricularia auricula-judae might be used to inhibit the differentiation of 3T3-L1 preadipocytes and reduction of triglycerides accumulation.

The aim of this study was to investigate the antitumor activity of solvent fractions from Auricularia auricula-judae 70% ethanol extract and confirmed the active components of dichloromethane fraction showing a potent antitumor activity than other fractions in the broncheoalveolar and gastric cancer cells. The solvent fractions of Auricularia auricula-judae extract, inhibited the growth proliferation of tumor cells in dose-dependent manner. The principle components of dichloromethane fraction were 5,11,17,23-tetrakis (1,1-dimethyl)-28-methoxypentacyclo [19.3.1.1 (3,7).1 (… (65.85%) and diazane (6.17%). The antitumor active components, diazane and gibberellic acid (GA3) were identified in this fraction by GC-MS analysis and lower antitumor activities than dichloromethane fraction. The unknown components of dichloromethane fraction were responsible for its cytotoxic effects on tumor cells. Based on IC50 value, gibberellic acid was little cytotoxic itself. According to PCR amplification, the apoptosis of tumor cells were induced by the down-regulation of Bcl-2 and over-expression of P53 on the presence of solvent fractions, diazane and gibberellic acid. Thus, these findings suggest that the dichloromethane might be used as functional feed additive that suppress the tumor growth in the body than other solvent fractions of Auricularia auricula-judae extracts.

Optimization study of the hot water extraction for enhancing antioxidative activity from Auricularia auricula was performed by Taguchi approach using orthogonal matrix L9(34) method. The correlation between DPPH radical scavenging activity and the components of samples extracted from different extraction conditions were also analyzed. The correlation coefficient between DPPH radical scavenging activity and melanin content of A. auricula was 0.93, indicating 'good correlation'. The optimum extraction conditions were obtained at the extraction time of 1 hr. temperature of 85oC, solid: water ratio of 1: 40(w/v) and frequency of 2 times. Under these conditions, values of maximum DPPH free radical scavenging activity and melanin contents of A. auricula were 67.21±2.17 and 52.94±2.10 mg/g, respectively. Melanin content of 1.6 times and DPPH free radical scavenging effect of 130% were enhanced by optimization.

The polysaccharides from fruit body of Auricularia auricula and Tremella fuciformis were extracted using hot water, and partially purified through ethanol precipitation and dialysis. Free radical scavenging activities of the crude and purified polysaccharides were examined and compared each other. Free radical scavenging activities of the partially purified polysaccharides were higher than those of crude polysaccharides. DPPH free radical, ABTS radical and SOD-like activities of partially purified polysaccharide at 1 mg/mL of concentration from A. auricula were 61.7, 9.6 and 38.9%, respectively, while those of T. fuciformis were 9.6, 5.7 and 15.3%, respectively. Results of site and nonsite specific hydroxyl radical scavenging activities indicated that the partially purified polysaccharide fractions from A. auricula and T. fuciformis exhibited the hydroxyl radical scavenging effect by hydrogen donating ability and iron ion chelating ability. Also, reducing powers of A. auricula and T. fuciformis were 77.1 and 14.7% of BHT (0.1%) as standard, respectively. It was suggested that antioxidant activities of A. auricula were about 1.4~6.4 times higher than those of T. fuciformis due to different levels of polyphenol content.

Auricularia auricula-judae has long been used as food and traditional remedies in Asian countries such as Korea and China. In this study, we evaluated the in vitro anti-tumor activity of various fractions from the ethanol extracts of Auricularia auricula-judae using various tumor cell lines. To do this, the mesh of Auricularia auricula-judae was mixed with 70% ethanol and heated at 1000C for 6 hrs and ethanol extract (ETOH) was collected. Ethanol extract was fractionated with dichloromethane (DCM), ethyl acetate, n-butanol and a water extract at room temperature as well as concentrated in a vacuum concentrator at a controlled temperature(<500C). The P388D1 macrophage and Sarcoma 180, human NSCLC NCI H358 (bronchioalveolar) and SNU1 cells (Gastric carcinoma) were cultured in RPMI. As the results, the cytotoxicity of the fractional extracts decreased significantly (P<0.05) in a dose-dependent manner. Dichloromethane extract (1 mg/ml) was the highest (P<0.05) in all experimental cell lines. There was also a significantly different sensitivity (P<0.05) among the P388D1, Sarcoma 180, NCI H358 and SNU1 cells for the fractional extracts. According to IC50 values, the most potent cytotoxic activity of dichloromethane fraction was found in Sarcoma 180 and NCI H358 cell lines. Butanol fraction appeared more cytotoxic to SNU1 cell line and water fraction had the highest cytotoxicity in P388D1 cell line. We did not find any significant difference between MTT and SRB assays in their ability to estimate cytotoxicity in all cell lines. Our findings suggest a potent antitumor activity of various fractions from the ethanol extracts of Auricularia auricula-judae depending on the solvent fractions and tumor cell lines. Further in vitro and in vivo studies will provide more information on the active compounds responsible for these activities and their potential as an anti-cancer remedy.

Phenotypic traits (physiological characteristics and somatic incompatibility) and genotypic traits (target region amplification polymorphism TRAP) were used to survey the diversity in Chinese Auricularia auricula systematically, which consisted of 32 main cultivated strains. The 27 important and stable physiological indexes were evaluated; somatic incompatibility test (SIT) reaction was described from three aspects: type, pigment, and intensity while intensity clustering alone revealed the phenotypic diversity among the 32 strains;16 pairs of TRAP primer combinations produced 535 unambiguous and reproducible DNA fragments, of these 524 (97.9%) were polymorphic. The phylogenetic trees were constructed by Unweighted Pair-group Method with Arithmetic Averages (UPGMA), which distributed the test strains into four to six major groups respectively. The principal coordinate analysis (PCO) of the three methods (physiological characteristics, SIT intensity and TRAP) exhibited highly similar clustering patterns, revealing that all the test strains can be divided into three groups considered significantly correlated with geographical regions. Results revealed the occurrence of relatively low diversity of A. auricula in the study. The cultivated strains in the same region are more similar physiologically and some strains are suspected to be synonymous. This study suggests that the value for estimating diversity are represented by TRAP＞SIT reaction intensity＞physiological characteristics in A. auricula, and PCO analysis can provided more effective and visible information than the UPGMA clustering dendrogram. Our finding will facilitate future germplasm resources management and breeding program of A. auricula in China.

A dark brownish pigment from fruit body of Auricularia auricula was isolated and characterized in this report. The pigment was obtained with a yield of 0.61%(w/w) by alkaline extraction and subsequent purification steps. It showed the positive FeCl3 test which was the indication of phenolic compounds. A synthetic melanin showed a similar spectrometric characteristics to the pigment extract regarding a characteristic UV absorption between 200-250 nm and infrared absorptions profiles in the finger print region including absorption peaks at 1701 and 1624 cm-1. Its element analysis indicated that its atomic copmposition is close to that of DOPA melanin (eumelanin). With the result of its antioxidant activity in the TNBT (5-thio-2-nitrobenzoic acid) assay, we concluded that the dark brownish pigment from A. auricula is a melanin-like compound having a powerful antioxidative activity.

흑목이 버섯의 자실체로부터 열수추출에 의해 얻은 조다당 분획물은 흑갈색 색소를 함유하였으므로 활성탄에 의한 탈색공정이 검토된 바 있다. 하지만 활성탄에 흡착된 색소 성분의 회수 및 활성탄의 재사용이 필요하였으므로 용매추출법을 이용한 용출 실험을 실시하였다. 증류수, 중성용매(3종) 및 알칼리 용매(4종) 등 8종 용매에 의한 1단 침출 실험 결과, 색소성분의 용해성은 1 M KOH 용액이 가장 우수하였다. 이러한 최적 용매인 1M KOH 용액을 이용하여 용액의 부피 및 시간에 따른 용출효과를 조사한 결과, 용액의 부피는 활성탄 1 g당 45 mL에서 최적의 용출량을 보였으며, 10분 이내에 빠르게 용출평형에 도달하였고, 용출공정의 용출속도는 2차 속도식으로 나타낼 수 있었다. 또, 25oC에서 1 M KOH 용액에 의한 다단침출의 실험 결과, 7단계 침출에서 최종 용출효율은 88.9%이었다. 그러나 온도가 증가할수록 용출효율이 급격히 높아져 95oC에서는 단일용출만으로 82.6%의 용출효율을 보였다. 한편, 용출 후의 재사용된 활성탄의 흡착효율(75.3%)은 신선한 활성탄(78.5%)과 거의 비슷하였고, 다당 정제율도 거의 비슷하여(1.21-1.25배) 재생 가능함을 보였다. 따라서, 흑목이 버섯의 흑갈색 성분의 탈색 후 흡착된 활성탄으로부터 용매추출을 이용한 효율적인 색소성분의 회수 및 활성탄 재생 공정의 가능성이 확인되었다.

흑목이 버섯의 자실체로부터 열수추출에 의해 흑갈색 색소를 함유한 조다당(crude polysaccharide)을 분획하였고, 이 조다당 분획의 흑갈색 색소 성분을 탈색 제거하기 위해 활성탄을 이용한 흡착공정을 검토하였다. 흡착 평형은 10분 이내의 빠른 시간에 도달하였으며, 최적의 흡착 조건은 pH 7.0 및 온도 40oC에서 조다당 용액 1 mL당 활성탄 16.7 mg을 첨가하였을 때 얻어졌다. 이 때의 최대 흡착효율은 약 80%이었고, 색소 성분의 제거로 다당은 정제되어 1.25배의 정제율을 나타내었다. 한편, 활성탄에 의한 색소의 등온흡착은 Langmuir 등온식에 따랐으며, 흡착속도는 2nd order model 식으로 나타낼 수 있었다. 또, 흡착은 확산을 통해 조절되는 물리적 흡착공정인 것으로 나타났다. 흑목이 버섯 조다당으로부터의 활성탄에 의한 색소흡착 공정은 그동안 보고된 바 없으며, 경제적이고 간편하여 향후 흑목이 버섯 조다당의 색소 제거와 정제 및 이에 의한 다당 정제공정으로서의 가능성이 있는 것으로 판단하였다.

본 연구에서는 젖산발효를 통해서 얻어진 목이버섯과 녹각 추출액 혼합물의 항산화 및 생리활성평가를 수행하였다. 녹각 추출액의 GABA생성 최적조건에서 probiotics 및 기능성 강화를 위해 목이버섯을 2.5% 첨가하여 30℃에서 7일간 젖산발효를 하였다. 발효 7일 pH 5.06, 산도 0.77%로 나타났으며 1.3×108 CFU/mL로 높은 균수를 유지하였고 GABA를 1.4% 생성하는 것으로 나타났다. 목이버섯을 첨가했을 때 젖산균 발효물의 물성이 개선되며 단기간에 고농도의 GABA를 생성하는 것으로 나타났다. 녹각 추출액의 젖산발효물의 세포 생존율을 실험한 목이버섯 2.5% 조건에서 발효 전 6 mg/mL에서 독성이 나타났으나 발효 후 독성이 완화되는 것으로 나타났다. 발효 후 6 mg/mL 농도에서는 5.58 μM로 발효 후 NO 생성이 감소되는 것으로 나타났다. 결론적으로 녹각 추출액과 목이버섯 혼합물의 젖산균을 이용한 정치배양을 통해 단기간에 고농도의 GABA 생산이 가능하였으며, 젖산 발효물은 세포독성 완화 효과 및 NO 생성을 저해하는 것으로 나타났다. 따라서 발효물은 GABA, probiotic, 식이섬유 등을 함유하여 기능성 식품소재로 이용이 가능할 것으로 기대된다.

In order to produce the high quality of dried-ear mushroom, various drying methods such as hot-air drying at 40~80˚C, freeze drying and drying in vinyl house were carried out. Drying hours of hot-air drying, freeze drying and drying in vinyl house were 12.5~21.5, 36.0 and 72.0 hrs, respectively. Vitamin D2 content of sample was the highest as 6.77μg/g DW in drying in vinyl house and then followed by freeze drying as 5.90μg/g DW and hot-air drying as 1.89~5.01μg/g DW. After dry, external appearance and color of mushrooms applied hot-air drying and drying in vinyl house were better than freeze-dried one. After rehydration, water uptake of sample in drying in vinyl house and hot-air drying at 50~60˚C were 17.8 and 19.3~21.0 times, respectively. The methods of drying in vinyl house and hot-air drying at 50~60˚C also led to high hardness, good shape and resilience. As the results of production of dried-ear mushroom with high quality, we suggest that the best method for drying is the drying in vinyl house due to not only high vitamin D2 content, good external appearance and color after drying but also high hardness and good shape after rehydration.