신령버섯의 기형으로부터 배양불능세균의 PCR 검정

신평균 외 한국버섯학회 2010 한국버섯학회지 Vol.8 No.4

신령버섯은 항암작용과 더불어 면역강화작용 등 약용으로서 이용되고 있습니다. 최근에는 신령버섯 재배농가에서 새로운 병징 현상이 보고되어 내생세균에 대한 분자생물학적 방법으로 조사하였다. 병징을 나타내는 수집된 기형버섯을 eubacterial 16S rDNA 영역을 이용하여 PCR한 결과 기형버섯만 증폭되었다. 이 영역을 부분적으로 염기서열을 결정한 결과 CFB bacterium과 유사성이 가장 높았고, 이 염기서열을 이용하여 프라이머를 디자인한 후 nested PCR를 부위별로 확인한 결과 기형을 일으킨 갓의 주름살 부위에서 가장 강하게 증폭되었고 포자수확도 되지 않았으며 배양불능세균 group인 CFB bacterium임을 확인하였다. Agaricus blazei Murill is a important medicinal mushroom for a powerful immune system builder and tonic. Currently, it is known about a new disease phenomenon that appears to be occurring on a number of mushroom farms. We described a straightforward approach in which molecular methods was used to survey the presence of potentially endo- and epiphytic bacteria infected with the Agaricus blazei. The 16S rDNA was amplified with universal eubacterial primers directly from pure cultures of Agaricus blazei mycelium and fruit body. The 16S rDNA sequences were almost identical (96 to 97% similarity), and phylogenetic analysis placed them in a single unique rRNA branch belong to the uncultural bacterium phylogroup. PCR detection of uncultural bacterium in the malformed tissues of Agaricus blazei were carried out by using 16S rRNA sequenced specific probe. It was strongly amplified at the malformed pileus region of fruit body and also spore print was impossible.

신령버섯의 기형으로부터 배양불능세균의 PCR 검정

신평균,박윤정,유영복,공원식,장갑열,오세종,이금희 한국버섯학회 2010 한국버섯학회지 Vol.8 No.4

Agaricus blazei Murill is a important medicinal mushroom for a powerful immune system builder and tonic. Currently, it is known about a new disease phenomenon that appears to be occurring on a number of mushroom farms. We described a straightforward approach in which molecular methods was used to survey the presence of potentially endo- and epiphytic bacteria infected with the Agaricus blazei. The 16S rDNA was amplified with universal eubacterial primers directly from pure cultures of Agaricus blazei mycelium and fruit body. The 16S rDNA sequences were almost identical (96 to 97% similarity), and phylogenetic analysis placed them in a single unique rRNA branch belong to the uncultural bacterium phylogroup. PCR detection of uncultural bacterium in the malformed tissues of Agaricus blazei were carried out by using 16S rRNA sequenced specific probe. It was strongly amplified at the malformed pileus region of fruit body and also spore print was impossible.

Bcl-2 and Caspase-3 Are Major Regulators in <i>Agaricus blazei</i>-Induced Human Leukemic U937 Cell Apoptosis through Dephoshorylation of Akt

Jin, Cheng-Yun,Moon, Dong-Oh,Choi, Yung Hyun,Lee, Jae-Dong,Kim, Gi-Young Pharmaceutical Society of Japan 2007 Biological & pharmaceutical bulletin Vol.30 No.8

<P><I>Agaricus blazei</I> is a medicinal mushroom that possesses antimetastatic, antitumor, antimutagenic, and immunostimulating effects. However, the molecular mechanisms involved in <I>A. blazei</I>-mediated apoptosis remain unclear. In the present study, to elucidate the role of the Bcl-2 in <I>A. blazei</I>-mediated apoptosis, U937 cells were transfected with either empty vector (U937/vec) or vector containing cDNA encoding full-length Bcl-2 (U937/Bcl-2). As compared with U937/vec, U937/Bcl-2 cells exhibited a 4-fold greater expression of Bcl-2. Treatment of U937/vec with 1.0—4.0 mg/ml of <I>A. blazei</I> extract (ABE) for 24 h resulted in a significant induction of morphologic features indicative of apoptosis. In contrast, U937/Bcl-2 exposed to the same ABE treatment only exhibited a slight induction of apoptotic features. ABE-induced apoptosis was accompanied by downregulation of antiapoptotic proteins such as X-linked inhibitor of apoptosis protein (XIAP), inhibitor of apoptosis protein (cIAP)-2 and Bcl-2, activation of caspase-3, and cleavage of poly(ADP-ribose)polymerase (PARP). Ectopic expression of Bcl-2 was associated with significantly induced expression of antiapoptotic proteins, such as cIAP-2 and Bcl-2, but not XIAP. Ectopic expression of Bcl-2 also reduced caspase-3 activation and PARP cleavage in ABE treated U937 cells. Furthermore, treatment with the caspase-3 inhibitor z-DEVD-fmk was sufficient to restore cell viability following ABE treatment. This increase in viability was ascribed to downregulation of caspase-3 and blockage of PARP and PLC-γ cleavage. ABE also triggered the downregulation of Akt, and combined treatment with LY294002 (an inhibitor of Akt) significantly decreased cell viability. The results indicated that major regulators of ABE-induced apoptosis in human leukemic U937 cells are Bcl-2 and caspase-3, which are associated with dephosphorylation of the Akt signal pathway.</P>

On-Line Estimation of Cell Growth from Agitation Speed in DO-Stat Culture of a Filamentous Microorganism, Agaricus blazei

Na, Jeong-Geol,Kim, Hyun-Han,Chang, Yong-Keun The Korean Society for Biotechnology and Bioengine 2005 Biotechnology and Bioprocess Engineering Vol.10 No.6

A simple, but effective on-line method for estimating the mycelial cell mass concentration from agitation speed data, a most readily-available process variable, has been developed for DO-stat cultures of Agaricus blazei. The dynamic change of dissolved oxygen concentration (DOC) in the initial transient period and the change in yield were considered in the development of the estimation algorithm or estimator. Parameters in the estimation algorithm were calculated from the agitation speed data at 20% of DOC. The proposed estimator could accurately predict the cell mass concentration regardless of DOC levels in the tested range of $10{\sim}40%$, showing a good extrapolation capability.

Methanol Extract of Agaricus blazei Murill Reduces Hepatic Damage Induced by $CCl_4$ and High Fat and Improves Liver Lipid Profile in Rats

Jung, Myung-Eun,Kwon, Hyuck-Se,Shin, Se-Gye,Jin, Yong-Xie,Han, Eun-Kyung,Ham, Seung-Shi,Kang, Il-Jun,Chung, Cha-Kwon The Korean Nutrition Society 2006 Nutritional Sciences Vol.9 No.4

The effect of methanol extract from Agaricus blazei Murill on the hepatotoxicity was investigated $CCl_4$ is one of the oldest and most widely used toxins for the induction of hepatic damages and fibrosis in experimental animals. In this study, male Sprague-Dawley(SD) rats were randomly divided into 6 groups of the control(C), $CCl_4(T),\;CCl_4$ and high fat group(TL) with matching sub-groups of Agaricus blazei Murill extract-fed groups of CA, TA and TLA. Methanol extracts of Agaricus blazei Murill were fed 50 mg/kg B.W daily via drinking water. A 1.2 mL of $CCl_4/kg$ body weight was administered by oral intubation twice a week for total of six times. The levels of total-cholesterol, TG, LDL and LDL-phospholipids were elevated by $CCl_4$ treatment as compared to the control(C). However, Agaricus blazei Murill methanol extract feeding in the group of TA and TLA significantly(p<0.05) decreased TG by 53.1 % and 17.9% compared to the internal control of T and TL, respectively. Triglyceride of TL was increased by 3.33 times(p<0.05) compared to the control(C) with $CCl_4$ and high fat administration from 3.78 mg/g to 12.60 mg/g liver. The extract(CA) also reduced kidney weight compared to the control(C). With the administration of high fat and $CCl_4$(TLA), the extract reduced the organ weight of both liver and kidney and further, significantly reduced TG, total cholesterol and GTP activity. Hepatoprotective effects of Agaricus blazei Murill on GOT, GPT, AP and LDH activities were enhanced by the extract feeding. Electronmicrograph showed that $CCl_4$ deteriorated the structure of cytoplasmic matrix with its uneven distribution. However, the extract reconstituted the damaged cytoplasm and stimulated mitochondriogenesis. The above results suggest that Agaricus blazei Murill may have a possible protective effect against chemically induced liver damage and further help to reduce the symptoms of fatty liver.

Methanol Extract of Agaricus blazei Murill Reduces Hepatic Damage Induced by CCl₄ and High Fat and Improves Liver Lipid Profile in Rats

Myung-Eun Jung,Hyuck-Se Kwon,Se-Gye Shin,Yong-Xie Jin,Eun-Kyung Han,Seung-Shi Ham,Il-Jun Kang,Cha-Kwon Chung 한국영양학회 2006 Nutritional Sciences Vol.9 No.4

??The effect of methanol extract from Agaricus blazei Murill on the hepatotoxicity was investigated CCl₄is one of the oldest and most widely used toxins for the induction of hepatic damages and fibrosis in experimental animals. In this study, male Sprague-Dawley(SD) rats were randomly divided into 6 groups of the control(C), CCl₄(T), CCl₄and high fat group(TL) with matching sub-groups of Agaricus blazei Murill extract-fed groups of CA, TA and TLA. Methanol extracts of Agaricus blazei Murill were fed 50 ㎎/㎏ B.W daily via drinking water. A 1.2 ㎖ of CCl₄/㎏ body weight was administered by oral intubation twice a week for total of six times. The levels of total-cholesterol, TG, LDL and LDL-phospholipids were elevated by CCl₄treatment as compared to the control(C). However, Agaricus blazei Murill methanol extract feeding in the group of TA and TLA significantly(p<0.05) decreased TG by 53.1% and 17.9% compared to the internal control of T and TL, respectively. Triglyceride of TL was increased by 3.33 times(p<0.05) compared to the control(C) with CCl₄and high fat administration from 3.78 ㎎/g to 12.60㎎/g liver. The extract(CA) also reduced kidney weight compared to the control(C). With the administration of high fat and CCl₄(TLA), the extract reduced the organ weight of both liver and kidney and further, significantly reduced TG, total cholesterol and GTP activity. Hepatoprotective effects of Agaricus blazei Murill on GOT, GPT, AP and LDH activities were enhanced by the extract feeding. Electronmicrograph showed that CCl₄deteriorated the structure of cytoplasmic matrix with its uneven distribution. However, the extract reconstituted the damaged cytoplasm and stimulated mitochondriogenesis. The above results suggest that Agaricus blazei Murill may have a possible protective effect against chemically induced liver damage and further help to reduce the symptoms of fatty liver.

Effects of Liquid Culture of Agaricus blazei Murill on Protein, Mineral Levels and Enzyme Activities in Rats

Koh, Jin-Bog,Kim, Jai-Young THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 2004 Journal of biomedical laboratory sciences Vol.10 No.2

This study was performed to investigate the effects of liquid culture of Agaricus blazei Murill on the weight gains, food efficiency ratios, serum protein and mineral levels, and serum enzyme activities in growing male rats. Sprague-Dawley rats (7 weeks old) were given four different types of diets for a succeeding period of 6 weeks, respectively: a normal diet group, a control diet group (normal diet + 15% lard + 0.5% cholesterol), a 30% or 40% A. blazei diet groups (control diet + 30% or 40% A. blazei in water) according to the levels of A. blazei supplementation. The body weight gains, food efficiency ratios, and the liver of the rats fed control diet, 30% or 40% A. blazei diets were significantly increased compared to rats fed the normal diet, but those of rats fed the 30% and 40% A. blazei diets were similar to those of rats fed the control diet. The concentrations of total protein, albumin, glucose, and hematocrit value in serum of rats fed the control diet, 30% or 40% A. blazei diets were similar to those of rats fed the normal diet. The concentrations of urea and creatinine in serum of rats fed the 30% or 40% A. blazei diets were similar to those of rats fed the control diet, but the urea of rats fed the 30% and 40% A. blazei diets were significantly decreased compared to rats fed the normal diet. The concentrations of Ca, P, Fe and Mg, and Ca^(++), K^(+), Na^(+) and Cl^(-) of rats fed the control diet, 30% or 40% A. blazei diets were similar to those of rats fed the normal diet. There were no differences in the activities of aspartate aminotransferase, alanine aminotransferase, γ-glutamyltranspeptidase and alkaline phosphatase in the serum among the experimental groups. In conclusion, the rats fed the A. blazei maintained normal protein and mineral levels, and enzyme activities of serum. But the A. blazei feeding could not decrease the body and liver weights in the rats fed high cholesterol diets.

Effects of Liquid Culture of Agaricus blazei Murill on Protein, Mineral Levels and Enzyme Activities in Rats

Jin-Bog Koh,Jai-Young Kim 대한의생명과학회 2004 Biomedical Science Letters Vol.10 No.2

This study was performed to investigate the effects of liquid culture of Agaricus blazei Murill on the weight gains, food efficiency ratios, serum protein and mineral levels, and serum enzyme activities in growing male rats. Sprague-Dawley rats (7 weeks old) were given four different types of diets for a succeeding period of 6 weeks, respectively: a normal diet group, a control diet group (normal diet + 15% lard + 0.5% cholesterol), a 30% or 40% A. blazei diet groups (control diet + 30% or 40% A. blazei in water) according to the levels of A. blazei supplementation. The body weight gains, food efficiency ratios, and the liver of the rats fed control diet, 30% or 40% A. blazei diets were significantly increased compared to rats fed the normal diet, but those of rats fed the 30% and 40% A. blazei diets were similar to those of rats fed the control diet. The concentrations of total protein, albumin, glucose, and hematocrit value in serum of rats fed the control diet, 30% or 40% A. blazei diets were similar to those of rats fed the normal diet. The concentrations of urea and creatinine in serum of rats fed the 30% or 40% A. blazei diets were similar to those of rats fed the control diet. but the urea of rats fed the 30% and 40% A. blazei diets were significantly decreased compared to rats fed the normal diet. The concentrations of Ca, P, Fe and Mg, and Ca??, K?, Na? and Cl? of rats fed the control diet, 30% or 40% A. blazei diets were similar to those of rats fed the normal diet. There were no differences in the activities of aspartate aminotransferase, alanine aminotransferase, γ-glutamyltranspeptidase and alkaline phosphatase in the serum among the experimental groups. In conclusion, the rats fed the A. blazei maintained normal protein and mineral levels, and enzyme activities of serum. But the A. blazei feeding could not decrease the body and liver weights in the rats fed high cholesterol diets.

신령버섯 균사체 액체배양액이 고콜레스테롤혈증 흰쥐의 지질대사에 미치는 영향

고진복,김재영 한국생명과학회 2004 생명과학회지 Vol.14 No.4

신령버섯 균사체 액체배양액이 고콜레스테롤 식이를 섭취한 흰쥐의 지질대사에 미치는 영향을 조사하고자, 생후 7주령의 숫쥐에 표준식이를 급여한 정상군, 표준식이에 15% 돈지와 0.5% 콜레스테롤을 첨가한 식이를 급여한 대조군, 대조식이에 신령버섯 균사체 액체배양액을 음료수에 30% 및 40%로 혼합 급여한 군(30% 및 40% 신령버섯군) 등 4군으로 나누어 6주간 사육한 결과는 다음과 같다. 실험동물의 체중 증가량, 식이섭취량 및 식이효율은 대조군과 30%와 40% 신령버섯군이 유사하였다. 간, 신장 및 부고환지방의 무게는 대조군과 신령버섯군들이 비슷하였다 간의 콜레스테롤 농도, 간과 혈청의 중성지질 농도는 대조군과 신령버섯군들이 비슷한 수준으로 감소효과가 나타나지 않았다. 혈청의 총 콜레스테롤과 LDL-콜레스테롤 농도 및 동맥경화지수는 대조군에 비해 30% 및 40% 신령버섯군이 유의하게 감소되었다. 총콜레스테롤에 대한 HDL-콜레스테롤의 비율은 대조군에 비해 신령버섯군들이 유의하게 증가되었다. HDL-콜레스테롤 농도는 대조군과 신령 버섯군들이 비슷한 수준으로 신령 버섯섭취에 따른 영향은 나타나지 않았다. 분으로 배설된 지방은 대조군에 비해 40% 신령버섯군이 유의하게 증가되었다. 이상의 결과로 보아 고콜레스테롤 식이를 급여한 흰쥐에 신령버섯 균사체 액체배양액을 30%나 40% 급여시 혈청의 총 콜레스테롤, LDL-콜레스테롤 농도 및 동맥경화지수를 낮추고, 총 콜레스테롤에 대한 HDL-콜레스테롤의 비율을 증가시키는 것으로 나타났다. The effects of liquid culture of Agaricus blazei Murill on the weight gains, serum and hepatic lipid concentrations were studied in male rats. Sprague-Dawley rats (7 weeks old) were given four different types of diets for 6 weeks, respectively: a normal diet group, a high fat control diet group (normal diet+15% lard +0.5% cholesterol), a 30% or 40% A. blazei diet groups (high fat control diet+30% or 40% A. blazei in water) according to the levels of A. blazei supplementation. The body weight gains, food intake, food efficiency ratios, and the liver, kidney, and epididymal fat pad weights of the rats fed 30% or 40% A. blazei diets were similar to those of the rats fed high fat control diet. The concentrations of serum total cholesterol, LDL-cholesterol and atherogenic index in rats fed the 30% or 40% A. blazei diets were significantly decreased compared with those of rats fed the high fat control diet. The HDL-cholesterol/total-cholesterol ratios of the rats fed 30% or 40% A. blazei diet were significantly increased compared with those of rats fed the high fat control diet. The fecal excretion of total lipid . in the rats fed 40% A. blazei diet was significantly increased compared with those of rats fed the high fat control diet. The concentrations of serum total lipid in the rats fed 40% A. blazei diet was significantly lower than that in the rats fed high fat control diet. But the concentrations of serum HDL-cholesterol, hepatic total cholesterol and triglyceride of rats fed the 30 or 40% A. blazei diets were similar to those of rats fed the high fat control diet. These results showed that the 30 or 40% A. blazei diets feeding decreased the total cholesterol, LDL-cholesterol and atherogenic index, and increased the HDL-cholesterol/total-cholesterol ratio in serum of rats.

Effects of Liquid Culture of Agaricus blazei Murill on Protein, Mineral Levels and Enzyme Activities in Rats

Koh, Jin-Bog,Kim, Jai-Young 대한의생명과학회 2004 Biomedical Science Letters Vol.10 No.2

This study was performed to investigate the effects of liquid culture of Agaricus blazei Murill on the weight gains, food efficiency ratios, serum protein and mineral levels, and serum enzyme activities in growing male rats. Sprague-Dawley rats (7 weeks old) were given four different types of diets for a succeeding period of 6 weeks, respectively: a normal diet group, a control diet group (normal diet + 15% lard + 0.5% cholesterol), a 30% or 40% A. blazei diet groups (control diet + 30% or 40% A. blazei in water) according to the levels of A. blazei supplementation. The body weight gains, food efficiency ratios, and the liver of the rats fed control diet, 30% or 40% A. blazei diets were significantly increased compared to rats fed the normal diet, but those of rats fed the 30% and 40% A. blazei diets were similar to those of rats fed the control diet. The concentrations of total protein, albumin, glucose, and hematocrit value in serum of rats fed the control diet, 30% or 40% A. blazei diets were similar to those of rats fed the normal diet. The concentrations of urea and creatinine in serum of rats fed the 30% or 40% A. blazei diets were similar to those of rats fed the control diet. but the urea of rats fed the 30% and 40% A. blazei diets were significantly decreased compared to rats fed the normal diet. The concentrations of Ca, P, Fe and Mg, and $Ca^{++}$, $K^+$, $Na^+$ and $Cl^-$ of rats fed the control diet, 30% or 40% A. blazei diets were similar to those of rats fed the normal diet. There were no differences in the activities of aspartate aminotransferase, alanine aminotransferase, $\gamma$-glutamyltranspeptidase and alkaline phosphatase in the serum among the experimental groups. In conclusion, the rats fed the A. blazei maintained normal protein and mineral levels, and enzyme activities of serum. But the A. blazei feeding could not decrease the body and liver weights in the rats fed high cholesterol diets.