• 제목/요약/키워드: Submerged culture

검색결과 278건 처리시간 0.025초

Monascus sp. KS 2 로 부터 색소 생산 변이주의 분리와 배양조건의 최적화 (Isolation of Pigment-Producing Mutants from Monascus sp. KS2 and Optimization of Cultural Conditions)

  • 박형은;김천호;민경희
    • 한국균학회지
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    • 제19권2호
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    • pp.120-127
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    • 1991
  • 식품용 색소로 사용되고 있는 Monascus-pigment(홍국색소) 를 산업적으로 생산하기 위하여 공기, 토양등으로부터 균주를 순수분리하여 색소생산용 배지에서 $30^{\circ}C$로 5일 동안 액체배양한 결과 색소생산이 가장 높은 Monascus sp. KS 2 를 분리하였다. 이균주를 모균주로 하여 NTG로 돌연변이를 유발한 결과 6개의 돌연변이주를 얻었으며, 색소생산성을 비교한 결과 가장 강력한 색소생산성 돌연변이주인 Monascus sp. SUR 37 을 선별하였으며, 모균주에 비해 적색색소와 황색색소가 각각 2.4배, 1.6배 높았다. 색소생산의 최적배양 조건은 온도 $30^{\circ}C$, pH6.0, 탄소원으로는 rice powder 5%, 질소원으로는 monosodium glutamate 0.15%, 그리고 아미노산으로는 DL-threonine, 배양기간은 4일이 가장 효과적이었다.

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Hypoglycemic and Hypocholesterolemic Effects of Botryosphaeran from Botryosphaeria rhodina MAMB-05 in Diabetes-Induced and Hyperlipidemia Conditions in Rats

  • Miranda-Nantes, Carolina C.B.O.;Fonseca, Eveline A.I.;Zaia, Cassia T.B.V.;Dekker, Robert F.H.;Khaper, Neelam;Castro, Inar A.;Barbosa, Aneli M.
    • Mycobiology
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    • 제39권3호
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    • pp.187-193
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    • 2011
  • Botryosphaeran, a water-soluble exopolysaccharide of the ${\beta}-(1{\rightarrow}3;1{\rightarrow}6)$-D-glucan type that has been isolated from the culture medium of Botryosphaeria rhodina MAMB-05 grown in submerged fermentation using glucose as the sole carbon source, was previously demonstrated to be non-genotoxic in peripheral blood and bone marrow, and exhibited strong anticlastogenic activity. In the present study, the effects of botryosphaeran were investigated in streptozotocin-induced diabetic rats as well as in high-fat diet-fed hyperlipidemic Wistar rats. The plasma glucose level was reduced by 52% in the diabetic group of rats after administration of 12 mg botryosphaeran/kg body weight of the rats (b.w.)/day by gavage over 15 days. A reduction in the median ration intake was accompanied by an increase in the median body weight gain, as well as the efficiency of food conversion. These results demonstrate that botryosphaeran has protective effects by reducing the symptoms of cachexia in Diabetes mellitus. Botryosphaeran administered by gavage at a concentration of 12 mg botryosphaeran/kg b.w./day over 15 days also reduced the plasma levels of total cholesterol and low density lipoprotein-cholesterol by 18% and 27%, respectively, in hyperlipidemic rats. Based on these findings, we conclude that botryosphaeran possesses hypoglycemic and hypocholesterolemic properties in conditions of diabetes mellitus and hyperlipidemia, respectively, and may be used as an oral anti-diabetic agent.

Evaluation of Ten Wild Nigerian Mushrooms for Amylase and Cellulase Activities

  • Jonathan, Segun Gbolagade;Adeoyo, Olusegun Richard
    • Mycobiology
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    • 제39권2호
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    • pp.103-108
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    • 2011
  • Amylases and cellulases are important enzymes that can be utilized for various biological activities. Ten different wild Nigerian mushrooms (Agaricus blazei, Agaricus sp., Corilopsis occidentalis, Coriolus versicolor, Termitomyces clypeatus, Termitomyces globulus, Pleurotus tuber-regium, Podoscypha bolleana, Pogonomyces hydnoides, and Nothopanus hygrophanus) were assayed for production of these secondary metabolites. The results revealed that most of the tested wild fungi demonstrated very good amylase and cellulase activities. With the incorporation of carboxymethyl-cellulose (a carbon source) into the culture medium, Agaricus blazei had the highest amylolytic activity of 0.60 unit/mL (at $25^{\circ}C$, pH 6.8). This was followed in order by P. tuber-regium and Agaricus sp. with 0.42 and 0.39 unit/mL, respectively ($p {\leq} 0.05$). Maltose and sucrose supplementation into the submerged liquid medium made N. hygrophanus and P. hydnoides to exhibit very low amylase activities of 0.09 and 0.11 unit/mL, respectively. Introducing peptone (an organic nitrogen source) into the basal medium enhanced the ability of C. versicolor to produce a cellulase value of 0.74 unit/mL. Other organic nitrogen sources that supported good cellulase activities were yeast extract and urea. Sodium nitrate (inorganic nitrogen source) generally inhibited cellulase production in all mushrooms. The best carbon source was carboxymethyl-cellulose, which promoted very high cellulase activity of 0.67 unit/mL in C. versicolor, which was followed in order by P. tuber-regium, T. chypeatus, and C. occidentalis ($p {\leq} 0.05$). Sucrose was the poorest carbon compound, supporting the lowest values of 0.01, 0.01, and 0.14 unit/mL in P. hydnoides, A. blazei, and Agaricus sp., respectively.

Investigation of Filamentous Fungi Producing Safe, Functional Water-Soluble Pigments

  • Heo, Young Mok;Kim, Kyeongwon;Kwon, Sun Lul;Na, Joorim;Lee, Hanbyul;Jang, Seokyoon;Kim, Chul Hwan;Jung, Jinho;Kim, Jae-Jin
    • Mycobiology
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    • 제46권3호
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    • pp.269-277
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    • 2018
  • The production of water-soluble pigments by fungal strains indigenous to South Korea was investigated to find those that are highly productive in submerged culture. Among 113 candidates, 34 strains that colored the inoculated potato dextrose agar medium were selected. They were cultured in potato dextrose broth and extracted with ethanol. The productivity, functionality (radical-scavenging activities), and color information (CIELAB values) of the pigment extracts were measured. Five species produced intense yellowish pigments, and two produced intense reddish pigments that ranked the highest in terms of absorbance units produced per day. The pigment extracts of Penicillium miczynskii, Sanghuangporus baumii, Trichoderma sp. 1, and Trichoderma afroharzianum exhibited high radical-scavenging activity. However, the S. baumii extract showed moderate toxicity in the acute toxicity test, which limits the industrial application of this pigment. In conclusion, P. miczynskii KUC1721, Trichoderma sp. 1 KUC1716, and T. afroharzianum KUC21213 were the best fungal candidates to be industrial producers of safe, functional water-soluble pigments.

Acidophilic Tannase from Marine Aspergillus awamori BTMFW032

  • Beena, P.S.;Soorej, M.B.;Elyas, K.K.;Sarita, G. Bhat;Chandrasekaran, M.
    • Journal of Microbiology and Biotechnology
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    • 제20권10호
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    • pp.1403-1414
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    • 2010
  • Aspergillus awamori BTMFW032, isolated from sea water, produced tannase as an extracellular enzyme under submerged culture conditions. Enzymes with a specific activity of 2,761.89 IU/mg protein, a final yield of 0.51%, and a purification fold of 6.32 were obtained after purification through to homogeneity, by ultrafiltration and gel filtration. SDS-PAGE analyses, under nonreducing and reducing conditions, yielded a single band of 230 kDa and 37.8 kDa, respectively, indicating the presence of six identical monomers. A pI of 4.4 and a carbohydrate content of 8.02% were observed in the enzyme. The optimal temperature was found to be $30^{\circ}C$, although the enzyme was active in the range of $5-80^{\circ}C$. Two pH optima, pH 2 and pH 8, were recorded, although the enzyme was instable at a pH of 8, but stable at a pH of 2.0 for 24 h. Methylgallate recorded maximal affinity, and $K_m$ and $V_{max}$ were recorded at $1.9{\times}10^{-3}$M and 830 ${\mu}Mol$/min, respectively. The impacts of a number of metal salts, solvents, surfactants, and other typical enzyme inhibitors on tannase activity were determined in order to establish the novel characteristics of the enzyme. The gene encoding tannase, isolated from A. awamori, was found to be 1.232 kb, and nucleic acid sequence analysis revealed an open reading frame consisting of 1,122 bp (374 amino acids) of one stretch in the -1 strand. In silico analyses of gene sequences, and a comparison with reported sequences of other species of Aspergillus, indicate that the acidophilic tannase from marine A. awamori differs from that of other reported species.

발효소시지 제조에 적합한 스타터 선발 (Screening of Lactic Acid Bacteria as a Starter Culture in Fermented Sausage)

  • 유선아;서승호;박성은;손홍석
    • 한국식품영양과학회지
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    • 제43권8호
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    • pp.1289-1295
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    • 2014
  • 본 연구에서는 전통발효식품 13종과 천연물 4종에서 27균주를 분리한 후 생육속도가 빠르고 pH 저하 능력이 우수한 3종의 균주를 1차 선발하였으며, 분자생물학적 방법을 이용하여 동정한 결과 Staphylococcus warneri, Sta. epidermidis, Lactobacillus plantarum과 99% 상동성을 보였다. 선발된 3균주의 발효소시지 스타터로서 이용가능성을 알아보기 위해 발효소시지의 환경과 유사한 model-system에서 배양하며 pH 저하 능력, 총산 생성 능력, 생육 능력, 아질산염 소거 능력을 발효소시지의 스타터로 많이 사용되고 있는 5균주 및 상업용 2균주와 비교하였다. Modelsystem에서 pH 저하능과 총산 생성능, 생육능은 관련성이 있었으며, 상업용 균주보다 분리한 Sta. epidermidis DO 10-1, Lac. plantarum MLK 14-2가 우수한 결과를 나타내었다. 아질산염 소거능의 경우에도 분리한 3균주가 상업용 균주보다 상대적으로 빠른 속도를 보였다. 분리한 3균주는 발효소지지 스타터로서의 발효 소거 능력은 우수할 것으로 보이지만 Staphylococcus는 잠재적인 위험성이 제기되는 균주이므로 Lac. plantarum이 발효소지지 제조에 가장 적합할 것으로 사료된다.

Antifibrotic Effect of Extracellular Biopolymer from Submerged Mycelial Cultures of Cordyceps militaris on Liver Fibrosis Induced by Bile Duct Ligation and Scission in Rats

  • Nan, Ji-Xing;Park, Eun-Jeon;Yang, Byung-Keun;Song, Chi-Hyun;Ko, Geonil;Sohn, Dong-Hwan
    • Archives of Pharmacal Research
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    • 제24권4호
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    • pp.327-332
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    • 2001
  • The antifibrotic effects of hot water extract (WEC), intracellular biopolymer (IPC) and extracellular biopolymers (EPC) from mycelial liquid culture of Cordyceps militaris on liver fibrosis were studied. Liver fibrosis was induced by a bile duct ligation and scission (BDL/S) operation, duration of 4 weeks in rats. In BDL/S rats, the levels of aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total bilirubin in serum and hydroxyproline content in liver were dramatically increased. The WEC or IPC treatment (30mg/kg/day for 4 weeks, p.o.) in BDL/S rats reduced the serum AST, ALT and ALP levels significantly (p<0.01). The EPC treatment (30 mg/kg /day for 4 weeks, p.o.) reduced the serum ALT, AST and ALP levels significantly (p<0.01). Malondialdehyde contents in liver treated with WEC, IPC or EPC were significantly reduced (p <0.05). But Liver hydroxyproline content was decreased only in EPC treated BDL/S rats to 55% that of BDL/S control rats (p < 0.01). The morphological characteristics and expression of alpha smooth muscle like actin in fibrotic liver, which appeared in BDL/S control group were improved in EPC treated fibrotic liver. These results indicate that IPC (30 mg/kg /day for 4 weeks, p.o.) has an antifibrotic effect on fibrotic rats induced by BDL/S.

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Antibacterial effect of urushiol on E. faecalis as a root canal irrigant

  • Kim, Sang-Wan;Shin, Dong-Hoon
    • Restorative Dentistry and Endodontics
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    • 제42권1호
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    • pp.54-59
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    • 2017
  • Objectives: The purpose of this study was to compare the antibacterial activity of urushiol against Enterococcus faecalis (E. faecalis) to that of NaOCl. Materials and Methods: The canals of thirty two single rooted human teeth were instrumented with Ni-Ti files (ProTaper Next X1, X2, X3, Dentsply). A pure culture of E. faecalis ATCC 19433 was prepared in sterile brain heart infusion (BHI) broth. The teeth were submerged in the suspension of E. faecalis and were incubated at $37^{\circ}C$ for 7 days to allow biofilm formation. The teeth were randomly divided into three experimental groups according to the irrigant used, and a negative control group where no irrigant was used (n = 8). Group 1 used physiologic normal saline, group 2 used 6% NaOCl, and group 3 used 10 wt% urushiol solution. After canal irrigation, each sample was collected by the sequential placement of 2 sterile paper points (ProTaper NEXT paper points, size X3, Dentsply). Ten-fold serial dilutions on each vials, and 100 µL were cultured on a BHI agar plate for 8 hours, and colony forming unit (CFU) analysis was done. The data were statistically analyzed using Kruskal-Wallis and Mann-whitney U tests. Results: Saline group exhibited no difference in the CFU counts with control group, while NaOCl and urushiol groups showed significantly less CFU counts than saline and control groups (p < 0.05). Conclusions: The result of this study suggests 10% urushiol and 6% NaOCl solution had powerful antibacterial activity against E. faecalis when they were used as root canal irrigants.

초자화동결된 생쥐 Preantral Follicle의 체외성장과 배란 (In vitro Follicular Growth and Ovulation of Mouse Preantral Follicles Cryopreserved by Vitrification)

  • 박지권;백원영
    • Clinical and Experimental Reproductive Medicine
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    • 제32권2호
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    • pp.91-99
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    • 2005
  • Objective: To define an appropriate vitrification condition of preantral follicle that yields high survival and to evaluate growth and ovulation rate of mouse follicles during in vitro culture after vitrification. Methods: Preantral follicles were isolated mechanically from mouse ovaries that were surgically recovered from mice aged 14 days. Retrieved preantral follicles were placed in EG (Ethylene Glycol) for 2, 5, 10 minutes and transferred to EFS-40 (40% EG, 18% Ficoll-70, 0.5 M sucrose) for 0.5, 1, 2 minutes. And then, preantral follicles were placed onto an EM grid and submerged immediately in liquid nitrogen. Thawing was carried out at room temperature. After defining the most appropriate vitrification condition that yields high survival, in vitro growth and ovulation rate of follicles were evaluated. Results: Appropriate vitrification condition that yield high survival rate ($83.2{\pm}2.1%$) of preantral follicle was EG for 5 minutes and EFS-40 for 0.5 minutes. In vitro survival rate of the vitrified preantral follicles were $85.5{\pm}0.5%$, $67.9{\pm}0.8%$ and $40.2{\pm}0.5%$ on day 2, 6 and 10. And in vitro growth of the vitrified preantral follicles were $107.1{\pm}16.1{\mu}m$, $117.1{\pm}18.4{\mu}m$, $178.4{\pm}45.6{\mu}m$ and $325.4{\pm}54.4{\mu}m$ on day 0, 2, 6 and 10. Although in vitro survival rate and growth of vitrified preantral follicles were lower than that of non-vitrified preantral follicles, the patterns of survival and growth were similar in vitrified and non-vitrified preantral follicles. The ovulation rate of antral follicles that was grown from vitrified preantral follicles was $32.6{\pm}1.2%$. Conclusion: Vitrified preantral follicles could be grown to antral sizes, and mature oocytes that can be used for IVF-ET programs were produced successfully. These data suggest that cryopreservation of preantral follicle by vitrification can be used for the preservation of the fertility.

db/db 마우스에서 Ceriporia lacerata 균사체 배양액이 GLUT4 발현에 미치는 영향 (Effect of submerged culture of Ceriporia lacerata mycelium on GLUT4 protein in db/db mouse)

  • 신은지;김지은;김지혜;박용만;윤성균;장병철;이삼빈;김병천
    • 한국식품저장유통학회지
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    • 제22권6호
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    • pp.893-900
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    • 2015
  • 최근 경제가 급속히 성장하면서 세계적으로 당뇨병이 심각한 대사성 질환으로 대두되고 있으며 이에 따라 혈당조절에만 초점을 맞춘 경구용 당뇨병 치료제가 아니라 당뇨병을 근본적으로 치료할 수 있는 혈당 강하 소재를 개발하기 위한 연구가 늘어나고 있다. 본 연구에서는 선행 연구에서 항당뇨 효능이 확인된Ceriporia lacerata 균사체 배양물건조물(CL01)을 제 2형 당뇨병 동물 모델인db/db 마우스에 경구투여하여 CL01이 혈액학적 지표 및 인슐린 신호 전달기전 상의 유관 단백질과 mRNA 발현에 미치는 영향을 평가하였다. CL01을 4주간 투여했을 때 혈당이 지속적으로 감소하였고, 혈청 인슐린, c-peptide 농도도 각각 음성대조군의 55.8%, 40% 수준으로 감소하였다(p<0.05). 인슐린 저항성을 반영하는 지표인 HOMA-IR 수치를 계산한 결과, CL01 투여군에서 인슐린 저항성이 음성대조군 대비 약 62% 개선되었다(p<0.05). 또한 인슐린 신호 전달에 관여하는 pIR, pAkt, pAMPK, GLUT4 단백질 및 Akt2, IRS1, GLUT4 mRNA 발현이 증가하였다. 이들 결과를 통해 CL01이 당 대사 및 인슐린 신호 전달에 관여하는 유관 단백질 및 유전자 발현에 영향을 미치며 이에 따라 인슐린 저항성을 개선하여 혈당을 효과적으로 감소시키는 것을 알 수 있다. 제 2형 당뇨병 환자의 90% 이상이 인슐린 저항성으로 인한 고인슐린혈증, 당내성 장애, 고혈압, 비만 등의 대사 이상이 생길 확률이 증가하기 때문에 CL01의 섭취를 통해 GLUT4 발현을 증가시켜 인슐린 저항성을 낮추는 것이 제 2형 당뇨병의 근본적인 치료에 이용될 수 있을 것으로 사료되며 이 연구가 당뇨병 치료의 추가적인 기전 연구를 위한 기초 증거로 활용될 수 있을 것이다.