We studied the effects of initial pH, different nitrogen sources, and cultivation methods (shake flask and static culture) on biomass production, exopolysaccharides (EPS), and adenosine by Paecilomyces tenuipes. Relatively low pH levels were optimal for mycelial growth and EPS production. Yeast extract was the most effective organic nitrogen source for EPS production, whereas soybean extract was the best for adenosine production. A high C/N ratio was beneficial for adenosine production; however, excessively high C/N ratios reduced adenosine production. Static fermentation significantly increased adenosine production. A Box-Behnken design was used to optimize adenosine production; the optimal conditions for adenosine production by P. tenuipes were pH 7.0, soybean concentration of 3%, and a static culture period of 20 days, with the maximum adenosine production of 141.10 mg/L (predicted value: 128.05 mg/L).
Cordycepin (3'-deoxyadenosine) is a nucleoside analog known for its diverse range of biological activities. This study investigated the effect of different types of sawdust on the production of the bioactive compound cordycepin. The results of the study showed that different types of wood sawdust affected the biosynthesis of cordycepin and a significant increase was observed when the conventional SDB medium was replaced with 1% NaOH treated pine sawdust. To optimize cordycepin production from Paecilomyces tenuipes in a medium containing 1% NaOH-pretreated pine sawdust, we employed Response Surface Methodology (RSM) in its Box-Behnken design (BBD) canonical form. The optimal conditions were determined as follows: a particle size of 109.5111-mesh (140 m) for 1% NaOH-pretreated pine sawdust, an input weight of 21.1679 g/L, and an incubation time of 73.8423 hours. According to our model, this combination is expected to yield a maximum cordycepin content of 896.1428 g/mL. Experimental validation of this prediction was performed using the suggested optimal conditions, resulting in an average cordycepin content of 922.6771 g/mL across three replicates, thus confirming the model's accuracy.
Roridin E, produced by Podostroma cornu-damae, is a mycotoxin with anticancer activity. To increase the content of roridin E, submerged culture conditions were optimized using response surface methodology. Three factors, namely, medium initial pH, incubation time and agitation speed were optimized using a Box–Behnken design. The optimum submerged culture conditions to increase the content of roridin E included a medium with an initial pH of 4.0, an incubation time of 12.90 days, and an agitation speed of 63.03 rpm. The roridin E content in the submerged culture, under the aforementioned conditions, was 40.26 mg/L. The findings of this study can help lower the current price of roridin E and promote its related research.
The choice of the culture medium is an important factor for the mass production of mycelia in submerged cultures. The influence of liquid medium on the mycelial dry weight of Paecilomyces tenuipes was investigated in this study. The regression equation is expressed as Y=-1292.94187+17.78612X1+18.92425X2+2.11464X3-0.019375X1X2-0.006276X1X3+0.008177X2X3- 0.070169X1 2-0.292175X2 2-0.008818X3 2, where Y represents the value of the mycelial dry weight (g/L), X1 is the particle size of wood sawdust in liquid medium (mesh), X2 is the concentration of the wood sawdust in liquid medium, and X3 is incubation time (h). The medium was optimized using a response surface methodology, and the optimal medium contained 30 g of wood sawdust (140 mesh), 20 g of glucose, and 10 g/L of peptone. Under these conditions, the mycelial dry weight reached 38.1 g/L (actual value). The culture medium containing wood sawdust is simple and easy to use, highly efficient, and eco-friendly, and its effectiveness in large preparations of P. tenuipes mycelia with low material costs has been demonstrated.
Destruxins (Dtxs) are insecticidal cyclic hexadepsipeptides produced by the entomopathogenic fungus Metarhizium anisopliae. Media composition for dtxs production was optimized with industrial grade media. Glycerol and casein peptone were selected as a carbon source and a nitrogen source, respectively. Dtxs production varied with C/N ratios. High yields of dtxs were observed at C/N ratios ranging 0.3 to 1.5, with concentrations mostly higher than 800 mg/L. Low yields were caused by high C/N ratio ranging from 3.0 to 8.0, resulting in less than 500 mg/L. The highest yield of Dtxs was obtained with 2% glycerol and 3% casein peptone, showing 192.2 mg/L of dtx A, 911.1 mg/L of dtx B, and 113.3 mg/L of dtx E, respectively. These results indicated that dtxs production is highly influenced by C/N ratio, especially the content of nitrogen source.
The entomopathogenic fungus Metarhizium anisopliae B is a powerful biological control agent against Monochamusalternatus, a crucial mediator of the pinewood nematode Bursaphelenchus xylophilus. In this study, production of destruxins(dtxs), insecticidal cyclic hexadepsipeptides, was monitored in the submerged culture of M. anisopliae B. Three typesof dtxs, i.e., destruxin A, B, and E, were produced during the culture. Among the three dtxs, the production yield ofdestruxin A was best, followed by destruxin B and E. Destruxin A production was increased when pH was controlledat 6.0, whereas production of destruxin E was not affected by the pH control. The highest yield of dtxs A, B, and Ewere 16.4, 7.3, and 6.1 mg L-1, respectively. Considering that process for dtxs production has not been optimized, M.anisopliae B has more powerful implication as a biocontrol agent.
Oxalic acid has a nematicidal activity against the root-knot nematode Meloidogyne incognita. High producer of oxalic acid was isolated, and then named as Aspergillus niger F22. Oxalic acid production was investigated under various temperatures from 20 – 33oC and rotational speeds in 5 L jar fermenters. Yield of oxalic acid increased with decreasing temperature. The highest yield was obtained at 23oC, showing the yield of oxalic acid of 8.7 g/L, whereas oxalic acid production was least at 33oC. At 20oC, the yield was lower than that of 23oC. At a rotational speed of 300 rpm, serious oxygen depletion was present from 48 - 72 h, resulting in low productivity of 26.2 mg /L·h. When a rotational speed was set at 600 rpm, dissolved oxygen tension was over 40% and oxalic acid production increased up to approximately 55%. Viscosity during the culture differed with temperatures. Viscosity increased with the increment of temperatures. When A. niger F22 was cultured at 23oC, viscosity was 810 cP, which was favorable for oxalic acid production.
A nematode, which showed entomopathogenicity, was isolated from Exomala orientalis in Korea, and then identified as Rhabditis blumi. Pathogenicity of R. blumi was evaluated against major cruciferous insect pests, including Artogeia rapae, Mamestra brassicae, and Plutella xylostella. In Petri-dish tests, insect mortality was dose and time dependent, which increased with dose (0–80 dauer juveniles/larva) and time increments. In greenhouse tests, P. xylostella larvae were most susceptible to nematodes, with insect reduction rate of 88.0 %. The rate varied with vegetable species and persistence time of live nematodes on vegetable leaves after spraying. The aeration rate was an important parameter for cultivation of the nematode R. blumi. A sufficient DOT level in the culture using an air-lift bioreactor should be maintained. The nematode growth rate increased with an increasing DOT level and/or bacteria uptake rate. The maximum nematode yield of 1.75 x 105 per ml was obtained under an aeration rate of 6 vvm.
Glycoproteins isolated from fruit bodies and mycelial cultures of mushrooms exhibit anti-carcinogenic actions in human cancer cells and animal tumor cells by induction of apoptosis. Here, we report that isoflavone-conjugated glycoproteins (designate Gluvone), exhibit strong anti-carcinogenic effects on human breast cancer MCF-7 cells by induction of apoptosis. Gluvone with 9.4 kDa of molecular weight was isolated from submerged-liquid culture of Agaricus blazei mycelia (ABM) in soy flake-containing liquid medium. MCF-7 cells were incubated with various amounts of Gluvone (0~250 μM) for a period of 6 days. Gluvone exhibited anti-proliferative actions in a dose-dependent manner and 62% growth inhibition at 200 μM for 4 days relative to control. Hoechst 33258 staining analysis revealed that Gluvone induced formation of apoptotic bodies. Gluvone was associated with down-regulation of anti-apoptotic Bcl-2 protein expression as well as up-regulation of pro-apoptotic Bax protein expression. Gluvone treatment induced proteolytic activation of caspase-9 and caspase-3 through cytochrome c release from mitochondria to cytosol as well as concomitant degradation of poly (ADP-ribose) polymerase (PARP). In addition, Gluvone induced activation of caspase-8. Taken all together, these results indicate that the anti-proliferative effect of Gluvone is associated with induction of apoptotic cell death through the mitochondrial dysfunction pathway mediated by enhancement of Bax protein expression and suppression of Bcl-2 protein expression.
본 연구는 mushroom complete medium(MCM) 액체배지에 수삼 추출물(GE, 65°Bx)을 첨가하여 면역활성이 증진된 노루궁뎅이버섯(Hericium erinaceum) 균사체를 배양하고, 균사체로부터 활성다당성분을 분획하고자 하였다. MCM에 대하여 GE를 5, 10과 15%(v/v) 첨가한 액체배지에서 균사체를 배양하고, 각각의 조다당획분(HE-GE-5-CP, HE-GE-10-CP와 HE-GE-15-CP)으로 분획하여 면역활성을 측정한 결과, HE-GE-10-CP는 HE-GE-5-CP와 HE-GE-15-CP보다 높은 활성을 나타내었으며, GE를 첨가하지 않은 MCM에서 배양된 균사체 조다당획분(HE-CP)보다 유의적으로 증진된 면역활성을 나타내었다. 또한, HE-GE-10-CP의 DEAE-Sepharose CL-6B 분획물 중 가장 높은 활성을 나타낸 HE-GE-10-CP-II획분은 대조군인 HE-CP의 어떠한 획분보다도 유의적으로 높은 면역활성과 암 전이 억제활성을 나타내었다. 한편, 활성획분인 HE-GE-10-CP-II는 arabinose, rhamnose, galactose, glucose와 uronic acid(molar ratio; 0.34:0.26:0.99:1.00:0.39)로 구성되어 있으나, 대조군인 HE-CP의 동일용매 용출획분으로서 HE-GE-10-CP-II보다는 활성이 낮은 HE-CP-II는 fucose, mannose, galactose와 glucose(molar ratio; 0.32:0.55:1.00:0.96)를 함유하여 다른 구성당 분포를 나타내었다. 따라서 노루궁뎅이버섯 균사체 액체배양에서 수삼 추출물 첨가는 균사체의 구성당 변화를 통한 면역활성 증진에 관여하는 것으로 사료되어 기능성 소재 개발에 유용할 것으로 사료된다.
In this study, we evaluated the antidiabetic effect of a submerged culture of Ceriporia lacerata mycelium (CL01) on hematological indices, as well as protein and mRNA expression of the insulin-signaling pathway, in db/db mice. After CL01 was administrated for 4 weeks, blood glucose levels decreased consistently, and plasma insulin and c-peptide levels each decreased by roughly 55.8%, 40% of those in the negative control (p<0.05). With regard to HOMA-IR, an insulin resistance index, insulin resistance of the CL01-fed group improved over that of the negative control group by about 62% (p<0.05). In addition, we demonstrated that the protein expression levels of pIR, pAkt, pAMPK, and GLUT4 and the mRNA expression levels of Akt2, IRS1, and GLUT4 in the muscle cells of db/db mice increased in the CL01-fed group compared to the corresponding levels in the control group. These results demonstrate that CL01 affects glucose metabolism, upregulates protein and gene expression in the insulin-signaling pathway, and decreases blood glucose levels effectively by improving insulin sensitivity. More than 90% of those who suffer from type 2 diabetes are more likely to suffer from hyperinsulinemia, hypertension, obesity, and other comorbidities because of insulin resistance. Therefore, it is possible that CL01 intake could be used as a fundamental treatment for type 2 diabetes by lowering insulin resistance, and these results may prove be useful as basic evidence for further research into the mechanisms of a cure for type 2 diabetes.
세리포리아 락세라타 균사체 배양물(CL01)의 혈당 강하 효과를 확인하기 위해 in-vitro 및 in-vivo 시험을 수행하였 다. CL01이 INS-1 세포에서 덱사메타손에 의한 세포 사멸 방지 효과를 나타냈으며 3T3-L1 세포에서는 당수송체인 GLUT4의 발현을 증가시켰다. 제 2형 당뇨 마우스를 4 그룹 (normal control (G1), negative control (G2), positive control (G3), CL01 250 mg/kg (G4))으로 나누어 6주간 매일 CL01 을 투여한 후 혈중 지표를 확인한 결과, CL01 섭취군은 체중, 사료 및 물 섭취량에서 음성대조군 대비 유의적인 차이를 보이지 않았다. 투여 5주 후 CL01 섭취군의 혈당이 음성대조군 대비 유의적인 감소를 보였으며 6주 경과 시 혈중 인슐린은 36% 증가하였고 혈중 C-peptide 농도는 18% 감소하였다. 경구 당 부하 시험 결과 CL01 섭취군의 혈당이 음성대조군 대비 15% 감소하였다. 이상의 결과를 통해 CL01이 베타 세포를 증식시켜 인슐린 분비를 촉진하고 혈 당을 낮추는 효과가 있음을 알 수 있다.
Sprassis crispa was cultivated using soybean curd whey, and its antimicrobial activities were examined against those of eight microorganisms that were foodborne pathogens or food-poisoning bacteria. The culture broth of soybean curd whey was superior in mycelium content (17.76 g/L) to that of the defined culture broth, and the β-glucan content was about 10.64 percent (w/w). The antimicrobial activities of the culture broth were confirmed against those of B. cereus, St. aureus, L. monocytogenes and S. typhimurium using the paper disk method. The antimicrobial activity was also maintained after the heat treatment and alcalase treatment. The filtrate with less than 3 kDa M.W. also showed the antimicrobial activity against four strains: B. cereus, St. aureus, L. monocytogenes and S. typhimurium. The minimum inhibitory concentration (MIC) was about 1.26 mg/mL in the B. cereus and 12.6 mg/mL in the St. aureus and L. monocytogenes. The S. typhimurium showed a MIC of 62.8 mg/mL. Thus, the culture of Sparassis crispa using soybean curd whey provides a thermally stable antimicrobial agent that can be used as a natural preservative in the biofood industry.
귤껍질배지에서의 소나무잔나비버섯(Fomitopsis pinicola) 균사체 배양에 따른 다당류의 함량과 세포의 형태학적 변화를 관찰하였다. 균사체의 수율이 가장 높은 배양 12일째의 생균사체 수율과 균사체의 알코올불용성물질의 함량은 YM 배지에서는 각각 11.29%(w/v, wet basis) 및 3.17%(w/w, wet basis)이었으나 CP배지에서는 각각 40.21%(w/v) 및 6.94%(w/w)였다. 그러나 산가용성 다당류는 YM배지
소나무 잔나비버섯 균사체 배양액(CM)을 이용한 빵 제조를 위하여 첨가량()에 따른 반죽의 발효 및 빵의 품질특성에 미치는 영향을 조사하였다. CM의 첨가량과 반죽의 품질지표(pH, 산도, 반죽부피, 되기, 강도 및 점도) 상호간의 상관분석을 행한 결과 CM은 반죽의 pH를 높임으로서 반죽의 되기 및 강도를 높이는 반면 점도를 감소시켰다. 첨가에 따라 굽기 손실률은 첨가에 따라 감소하였으나, loaf volume index는 첨가량에 따른 대조구