Strain Improvement by Overexpression of the LaeA Gene in Monascus pilosus for the Production of Monascus-Fermented Rice

( Sang Sub Lee ),( Jin Hee Lee ),( In Hyung Lee ) 한국미생물 · 생명공학회 2013 Journal of microbiology and biotechnology Vol.23 No.7

Monascus species have been used to produce fermented rice called Monascus-fermented rice (MFR). To improve a Monascus strain via activation of secondary metabolite (SM) gene clusters for use in the production of MFR, we overexpressed an ortholog of the laeA gene, which encodes a global positive regulator of secondary metabolism under the control of the strong heterologous Aspergillus nidulans alcA promoter in Monascus pilosus. The OE::laeA transformant produced more SMs, including those not detected under uninduced conditions. MFR produced using the M. pilosus OE::laeA strain contained 4 times more monacolin K, a cholesterol-lowering agent, than MFR produced using the wild-type strain. In addition, pigment production was remarkably increased, and the antioxidant activity was increased as well. The results from this study suggest that Monascus species, which are important industrial fermentative fungi in Asia, can be improved for the production of functional foods by overexpressing the laeA gene.

Monascus pilosus로 발효시킨 후발효 녹차가 고지방 식이 마우스의 체중과 혈청 지방함량 및 간 조직 항산화계 효소활성에 미치는 영향

이상일 ( Sang Il Lee ),이예경 ( Ye Kyung Lee ),김순동 ( Soon Dong Kim ),양승환 ( Seung Hwan Yang ),서주원 ( Joo Won Suh ) 한국응용생명화학회 2012 Journal of Applied Biological Chemistry (J. Appl. Vol.55 No.2

The anti-obese, hypolipidemic and hepatoprotective effects of post-fermented green tea by Monascus pilosus was tested with mice fed with high-fat diet for 7 weeks. The body weight gain and feed efficiency ratio (FER) in normal control group (NC), CHA (2% non-fermented green tea powder supplemented high-fat diet group) and mCHA (2% green tea powder post-fermented by M. pilosus supplemented high fat diet group) groups were significantly lower than those of high fat diet control group (HC). Epididymal fat weight in mCHA and NC were significantly lower than HC. The hepatic lipid peroxide was dramatically higher in HC than that of NC and was significantly lower in CHA and mCHA. In addition, dehydrogenase type activity of xanthine oxidoreductase in HC was lower than that of NC, but significantly higher than CHA and mCHA. In histopathological findings, hepatic fat accumulation in HC was higher than that of NC, CHA and mCHA. Antiobese, hypolipidemic and antifatty liver effect of green tea powder postfermented by M. pilosus was slightly higher than that of nonfermented green tea. In conclusion, the constituents of green tea fermented by M. pilosus has been proven to not only inhibit obesity and hyperlipidemia but also decrease the hepatic fat accumulation in high fat diet-induced obese mice.

Monascus pilosus로 발효시킨 비지의 항비만 효과

이상일 ( Sang Il Lee ),이예경 ( Ye Kyung Lee ),김순동 ( Soon Dong Kim ),이인애 ( In Ae Lee ),최종근 ( Jongkeun Choi ),서주원 ( Joo Won Suh ) 한국응용생명화학회 2014 Journal of Applied Biological Chemistry (J. Appl. Vol.57 No.1

This study investigated anti-obesity and antioxidant effects of dietary non-fermented soybean crud residue (SCR) and fermented SCR by Monascus pilosus (FSCR) in high-fat inducedobese mice. SCR and FSCR were supplemented with high-fat diet at 2% (wt/wt) dose for 8 weeks. Both SCR and FSCR significantly lowered body weight, epididymal fat weight and weight gain rate compared to high-fat diet control (HC) group and FSCR group showed lowest weight gain rate. In addition, it was observed that serum and hepatic lipid profiles including triglyceride, total cholesterol, LDL-cholesterol and HDL-cholesterol were significantly improved by supplementing SCR or FSCR. Furthermore, SCR and FSCR administration showed increase of glutathione content and decrease of hepatic lipid peroxide content, serum aminotransferase activity, and hepatic xanthine oxidase activity. On the other hand, activities of reactive oxygen species scavenging enzyme such as superoxide dismutase, glutathione S-transferase and glutathione peroxidase in two test groups were higher than those of HC. Lastly, in comparison with SCR, FSCR was more effective in restoring obesity-related biomarkers to normal level in high-diet induced obese mice. In conclusion, the present study indicates that FSCR could have not only anti-obese effects such as inhibition of abdominal fat accumulation, but also protective effects of cardiovascular disease such as atherosclerosis by decreasing serum and hepatic lipid contents. Furthermore, these results suggest that experimental diets in this study could alleviate hepatic damage caused by overproduction of reactive oxygen spices (ROS) due to obesity via inhibition of ROS generating activities and induction of ROS scavenging activities.

Monascus Pilosus로 발효시킨 검정콩 첨가 식이가 고지방식이 마우스의 체중과 혈청 지방함량 및 간 조직 항산화계 효소활성에 미치는 영향

이상일(Lee, Sang-Il),김순동(Kim, Soon-Dong),이예경(Lee, Ye-Kyung),김미정(Kim, Mee-Jung),이인애(Lee, In-Ae),최종근(Choi, Jongkeun),서주원(Suh, Joo-Won) 한국영양학회 2013 Journal of Nutrition and Health Vol.46 No.1

The anti-obesity effects of fermented black bean were tested with mice fed a high fat diet for seven weeks. Body weight gain and feed efficiency ratio (FER) in the high fat diet control (HC) group were markedly higher, compared with those of the normal control (NC) group, but were significantly lower in the 2% black bean powder supplemented high fat diet (BB) group and 2% black bean powder fermented by M. pilosus supplemented high fat diet (BBM) group, compared with those of the HC group. Food intake in the HC and BB groups was significantly lower than that of the NC and BBM groups. Water intake in the HC group was significantly lower than that of the NC group, but was higher in the BB and BBM groups, compared with that of the HC group. On the other hand, relative liver and kidney weight in the HC group was lower than that of the NC group, but was higher in the BB and BBM groups, compared with that of the HC group. In addition, whereas epididymal fat weight in the HC group was markedly higher than that of the NC group, it was significantly lower in the BB and BBM groups, compared with that of the HC group. Meanwhile, hepatic GSH in the HC group was significantly lower than that of the NC group, but was slightly higher in the BB and BBM groups, compared with that of the HC group. Although hepatic LPO in the HC group was dramatically higher than that of the NC group, it was significantly lower in the BB and BBM groups, compared with that of the HC group. In addition, serum TG, total cholesterol, and LDL-cholesterol in the HC group was significantly higher than that of the NC group, but was significantly lower in the BB and BBM groups, compared with that of the HC group. On the contrary, HDL-cholesterol in the HC group was significantly lower than that of the NC group, but was higher in the BB and BBM groups, compared with that of the HC group. In addition, activity of XOR D type in the HC group was lower than that of the NC group, but was slightly higher in the BB and BBM groups, compared with that of the NC group. Activities of ROS scavenging enzymes, such as SOD, GPX, and GST in the HC group were significantly lower than those of the NC group, but were significantly higher in the BB and BBM groups, compared with those of the HC group. In addition, serum ALT activity in the HC and BB groups was higher than that of the NC group, but was significantly lower in the BB and BBM groups, compared with that of the HC group. In histopathological findings, hepatic fat accumulation in the HC group was higher than that of the NC group, but was lower in the BBM group, compared with that of the HC and BB groups. In particular, antiobese, hypolipidemic, and antifatty liver effect of black bean powder fermented by M. pilosus was specifically higher than that of non-fermented steamed black bean. In conclusion, the constituents of black bean fermented by Monascus pilosus have been proven to not only inhibit obesity and hyperlipidemia but also decrease hepatic fat accumulation in high fat diet-induced obese mice. (Korean J Nutr 2013; 46(1): 5 ~ 14)

홍국균 발효가 발아현미의 Monacolin K 함량과 항산화 활성에 미치는 영향

이상훈(Sang Hoon Lee),장귀영(Gwi Yeong Jang),김민영(Min Young Kim),김신제(Shinje Kim),이연리(Yuon Ri Lee),이준수(Junsoo Lee),정헌상(Heon Sang Jeong) 한국식품영양과학회 2015 한국식품영양과학회지 Vol.44 No.8

발아 온도와 기간에 따른 발아현미 홍국배양물의 홍국색소, monacolin K, citrinin 및 페놀 화합물 생성량과 항산화 활성 변화를 조사하였다. 발아 기간이 증가함에 따라 명도는 증가하였으나 적색도와 황색도는 감소하였고, 적색(A500), 오렌지색(A470) 및 황색(A400) 색소 생성량도 동일한 경향이었다. 총 monacolin K는 현미(215.85 mg/kg)가 백미(40.41 mg/kg)보다 높았으며, 32°C에서 1일간 발아시킨 발아현미(1,263.04 mg/kg)에서 가장 높은 함량을 보였으나 발아 기간이 증가함에 따라 감소하였다. 곰팡이 독소인 citrinin은 모든 처리구에서 검출되지 않았다. 총 폴리페놀과 플라보노이드 함량은 발아 온도와 기간이 증가함에 따라 유의적으로 증가하여 37°C에서 4일간 발아시킨 발아현미에서 13.80 mg/g과 1.30 mg/g으로 가장 높았다. 전자공여능과 총 항산화력은 발아에 의해 증가되어 32°C에서 1일간 발아시킨 발아현미에서 각각 22.16 mg TE/g과 62.27 mg AAE/g으로 가장 높았으나 발아 기간이 증가함에 따라 감소하였다. 이상의 결과로부터 홍국색소와 monacolin K 생성량을 증가시키기 위한 기질로 발아현미가 유용하며, 배양에 사용된 Monascus pilosus 305-9는 citrinin을 생성하지 않고 monacolin K 생성량을 증가시켜 식품의 기능성 증진용 소재 및 균주로서의 활용이 기대된다. This study was performed to investigate the changes in monacolin K content and antioxidant activities of Monascus-fermented brown rice with different germination temperatures and periods. Brown rice was germinated at 32, 35 and 37°C for 1∼4 days, after white rice (WB), brown rice (BR), and germinated brown rice (GBR) were fermented with M. pilosus 305-9 at 30°C for 20 days. The redness, yellowness and Monascus pigments increased after germination. Total monacolin K content increased from 215.85 mg/kg of BR to 1,263.04 mg/kg of GBR (32°C/1 day), whereas monacolin K content decreased with increase in germination period. Citrinin was not detected in any of the samples. Total polyphenol (TPC) and flavonoid contents (TFC) increased with increase in germination temperature and period, whereas electron donating ability (EDA) and total antioxidant activities (TAA) decreased due to reduction of Monascus pigment content. The TPC and TFC showed the highest values (13.80 mg/g and 1.30 mg/g, respectively) in GBR (37°C/4 day), whereas EDA and TAA showed the highest values (22.16 mg Trolox equivalent/g and 62.27 mg ascorbic acid equivalent/g, respectively) in GBR (32°C/1 day). These results indicated that the optimal germination temperature and period for increasing monacolin K content and antioxidant activities was found to be at 32°C for 1 day. In addition, it was found that M. pilosus 305-9 was a useful strain for increasing monacolin K content without producing citrinin in functional foods and pharmaceutical industrial regions.

Antioxidant activity of Green Tea Fermented with Monascus pilosus

( Ye Kyung Lee ),( Sang Il Lee ),( Jeong Sook Kim ),( Seung Hwan Yang ),( In Ae Lee ),( Soon Dong Kim ),( Joo Won Suh ) 한국응용생명화학회 2012 Journal of Applied Biological Chemistry (J. Appl. Vol.55 No.1

Green tea leaves were fermented for 15 and 30 days with Monascus pilosus which is known to produce functional statins (TMs), and the content of various biochemical constituents such as total polyphenol (TP), total flavonoid (TF), theaflavin, and thearubigin were analyzed and compared with that of nonfermented green tea (GT) and Pu-erh Chinese post-fermented tea (PU). In addition to the electron donating ability (EDA), ferric iron reducing power (FIRP), xanthine oxidase (XO) inhibitory activity, superoxide dismutase (SOD)-like activity, iron chelating activity (ICA) and hydrogen peroxide contents were also measured and compared with that of GT and PU. Content of TP and TF in the water and ethanol extracts in TMs were lower than those in GT and PU. Theaflavin and thearubigin contents of water and ethanol extracts in TMs were higher than those of GT. And, these components were increased depending on the period of fermentation. While, EDA and FIRP of TMs were lower than those of GT, XO inhibitory activity of TMs was higher than nonfermented tea. While, ICA of TMs was slightly higher than GT and PU, the content of hydrogen peroxide in TMs was markedly lower than GT. This results suggested that the green tea fermented by M. pilosus was valuable for oxidative stress-induced diseases by decreasing hydrogen peroxide, and forming theaflavins and thearubigins with functionality of genus Monascus.

Monascus pilosus로 발효시킨 뽕잎차의 품질특성과 항산화능

이상일(Sang-Il Lee),이예경(Ye-Kyung Lee),최종근(Jongkeun Choi),양승환(Seung Hwan Yang),이인애(In-Ae Lee),서주원(Joo-Won Suh),김순동(Soon-Dong Kim) 한국식품영양과학회 2012 한국식품영양과학회지 Vol.41 No.5

Monascus pilosus에 의한 발효뽕잎차(FMM: fermented mulberry leaves tea)의 monacolin K 함량, citrinin의 유무, 관능적 품질, total polyphenol 및 total flavonoid의 함량, ROS 생성계 효소인 xanthine oxidase 저해활성 및 ROS 소거계 효소류의 활성을 건조뽕잎(UFM: unfermented mulberry leaves)과 비교하였다. FMM은 UFM에 비하여 색상기호도 및 종합적 기호도가 유의적으로 높았다. FMM에 함유된 total monacolin K 함량은 0.056%(dry basis)로 활성형이 83.93%를 차지하였으며 citrinin은 검출되지 않았다. Total polyphenol과 total flavonoid의 함량 모두 UFM에서 현저히 높았다. XO 저해활성과 SOD 유사활성은 농도에 비례하여 높았으며 UFM이 FMM에 비하여 높았다. 전자공여능과 ferric-reducing antioxidant power(FRAP)의 경우도 UFM과 FMM 모두에서 농도에 비례하여 증가하였으나, UFM에서 FMM에 비하여 현저히 증가하였다. Ferrous iron chelating 활성의 경우에도 추출물의 첨가 농도에 비례하여 증가하였고 그 증가의 정도는 UFM이 FMM에 비해 더욱 강하게 나타났으나, 4.0 ㎎/mL 이상의 농도에서는 감소하였다. 한편 polyphenol 또는 flavonoid 함량 당의 XO 저해활성과 SOD 유사활성 및 FRAP는 FMM이 UFM에 비하여 현저하게 높았다. 이상의 결과들을 종합해 볼 때, 뽕잎을 M.pilosus로 발효시킨 FMM은 홍국의 기능성을 가짐과 더불어 상당히 강한 항산화능과 기호성을 관찰할 수 있어 ROS로 야기될 수 있는 여러 질병의 예방과 치료에 도움을 줄 수 있는 기초자료가 될 것이라 사료된다. This study was carried out to investigate the contents of monacolin K and citrinin, along with the sensory quality and antioxidant activity of mulberry leaf tea fermented by Monascus pilsous (FMM). Total monacolin K content of FMM was 0.058%, but citrinin was not detected. Redness of brewed FMM was remarkably higher than that of unfermented mulberry leaf tea (UFM). In sensory evaluation of brewed FMM, while astringent taste and savory taste were lower, flavor, color, and overall acceptability were significantly higher than those of UFM. Total polyphenol contents of UFM and FMM were 83.1 and 23.61 ㎎/g (dry basis), total flavonoid contents of UFM and FMM were 17.96 and 3.99 ㎎/g (dry basis), respectively. Xanthine oxidase (XO) inhibitory activity and superoxide dismutase (SOD)-like activity of FMM were lower than those of UFM. Electron-donating ability and ferric-reducing antioxidant power of FMM were slightly lower than those of UFM. However, the antioxidant activities of FMM per polyphenol content were markedly higher than those of UFM. These results suggest that FMM may scavenge excessive reactive oxygen spices (ROS) via inhibition of XO and SOD-like activity. Furthermore, FMM demonstrated relatively higher acceptability and antioxidant ability along with functionality of Hongguk (red yeast rice), and therefore could be utilized to prevent various ROS-induced diseases.

Original Article: Food Science/Microbiology : Effect of Fermentation on the Antioxidant Activity of Rice Bran by Monascus pilosus KCCM60084

( Jin Hua Cheng ),( Bong Keun Choi ),( Seung Hwan Yang ),( Joo Won Suh ) 한국응용생명화학회 2016 Journal of Applied Biological Chemistry (J. Appl. Vol.59 No.1

In this study, we optimized fermentation conditions for the solid state fermentation of rice bran with Monascus pilosus KCCM60084, and the antioxidant activities were investigated. Optimal fermentation conditions were determined by the production of Monacolin K, a functional secondary metabolites with cholesterol lowering activity. The highest Monacolin K production were 2.88 mg/g observed on day 10 with 45% moisture content in the substrate when in℃ulated with 5% in℃ulum (w/w). Reducing power, iron chelating activity and ABTS+ radical scavenging activity were significantly enhanced after fermentation by 60, 80, and 38% respectively. Furthermore, the content of total flavonoid were found to be increased by 4.58 fold. Based on these results, Monascus-fermented rice bran showed strong possibility to be used as a natural antioxidant agent due to its enhanced antioxidant activity.

유산균 및 홍국균 발효 꽃송이버섯 추출물과 잔사의 식이섬유와 베타 글루칸의 함량

임창완(Chang Wan Lim),강경규(Kyoung Kyu Kang),유영복(Young-Bok Yoo),김병희(Byung Hee Kim),배송환(Song-Hwan Bae) 한국식품영양과학회 2012 한국식품영양과학회지 Vol.41 No.12

본 연구에서는 식이섬유와 β-glucan이 풍부한 꽃송이버섯을 발효한 후 추출하여 이들 성분의 함량을 보다 증가시킨 추출물과 잔사를 제조하고자 하였다. 이를 위해 열풍 건조한 꽃송이버섯 자실체의 분말을 유산균(L. brevis)과 홍국균(M. pilosus)으로 각각 발효한 후 열수와 수용성 에탄올(50, 70, 90%, v/v)로 추출하여 추출물과 잔사를 제조하고 이들의 수용성, 불용성 및 총 식이섬유와 β-glucan의 함량을 버섯원물의 추출물과 잔사의 각 성분의 함량과 비교하였다. 홍국균 발효 버섯의 총 식이섬유 함량은 74.4 g/100 g으로 버섯원물(64.4 g/100 g)과 유산균 발효 버섯(66.1 g/100 g)의 총 식이섬유 함량에 비해서 유의적으로(p<0.05) 높았다. 버섯원물, 유산균 발효 버섯, 홍국균 발효 버섯의 β-glucan 함량(21.9∼24.4 g/100 g) 간에는 유의적인 차이가 없었다. 추출물의 경우, 홍국균 발효 버섯의 열수 추출물에서 총 식이섬유(21.6 g/100 g)와 수용성 식이섬유 함량(19.3 g/100 g)이 가장 높았으며 버섯 원물의 열수 추출물의 총 식이섬유(16.4 g/100 g)와 수용성 식이섬유 함량(14.6 g/100 g)보다도 유의적으로(p<0.05) 높았다. 홍국균 발효 버섯을 열수 추출하고 남은 잔사의 총 식이섬유 함량은 90.5 g/100 g이었고 이들의 대부분이 불용성 식이섬유로 구성되어 있었으며, β-glucan 함량은 버섯 원물이나 홍국균 발효 버섯보다 높은 31.0 g/100 g이었다. 따라서 본 연구에서 제조한 홍국균 발효 꽃송이버섯의 열수 추출물과 잔사는 각각 액상과 분말 형태의 건강기능식품 및 가공식품 소재로 개발될 수 있을 것으로 기대된다. Sparassis (S.) crispa is an edible mushroom abundant in dietary fiber and β-glucan. The aim of this study was to prepare extracts and residues of the fruit bodies of S. crispa fermented with Lactobacillus (L.) brevis and Monascus (M.) pilosus and to measure the remaining dietary fiber and β-glucan. Dried powder of S. crispa containing 64.4 g/100 g total dietary fiber (2.6 g/100 g soluble and 61.8 g/100 g insoluble dietary fibers) and 24.0 g/100 g β-glucan was used as the starting material for the extraction. Raw and fermented S. crispa were extracted with hot water and three kinds of aqueous ethanol (50, 70, and 90%, v/v), respectively. A hot water extract from S. crispa fermented with M. pilosus had greater soluble dietary fiber content (19.3 g/100 g) than that from raw S. crispa with 14.6 g/100 g soluble dietary fiber or that from L. brevis-fermented S. crispa with 8.2 g/100 g soluble dietary fiber. The yield of the extract was 16.6% of intial weight of dried S. crispa. After hot water extraction of S. crispa fermented with M. pilosus, residues containing 90.5 g/100 g total dietary fiber (1.3 g/100 g soluble and 89.2 g/100 g insoluble dietary fibers) were obtained, and the yield was 69.6% of intial weight of dried S. crispa. The residue (31.0 g/100 g) contained more β-glucan than raw S. crispa or M. pilosus- fermented S. crispa (24.4 g/100 g). The resulting hot water extract and residue from S. crispa fermented with M. pilosus would be suitable for use in preparing liquid and powdered health functional foods, respectively.

Ameliorative Effects of Monascus pilosus-Fermented Black Soybean (Glycine max L. Merrill) on High-Fat Diet-Induced Obesity

Oh, Hong-Geun,Kang, Young-Rye,Lee, Hak-Yong,Kim, Jung-Hoon,Shin, Eun-Hye,Lee, Bong-Gun,Park, Sang-Hoon,Moon, Dae-In,Kim, Ok-Jin,Lee, In-Ae,Choi, Jongkeun,Lee, Ji-Ean,Park, Kwang-Hyun,Suh, Joo-Won 한국식품영양과학회 2014 Journal of medicinal food Vol.17 No.9

The purpose of this study was to examine the antiobesity effects of Monascus pilosus-fermented black soybean (F-BS) in C57BL/6 mice with high-fat diet (HFD)-induced obesity. F-BS (oral, 0.5 and 1.0 g/kg per body weight, twice per day) ameliorated obesity by reducing body and liver weight increases, and regulating blood glucose and cholesterol levels in C57BL/6 mice fed a control or HFD with oral administration of F-BS for 12 weeks. F-BS suppressed the growth of epididymal, retroperitoneal, and perirenal fat pads by preventing increases in the adipocyte size. Moreover, the levels of blood glucose, total cholesterol, and leptin were significantly lowered by F-BS administration in a dose-dependent manner. These results indicated that F-BS is a beneficial food supplement for preventing obesity, controlling blood glucose, and lowering cholesterol. Future research strategies should address the mechanisms that selectively regulate obesity, including hyperglycemia and hypercholesterolemia.