Paecilomyces tenuipes (P. tenuipes) is a fungus cultivated artificially by South Korean researchers, utilizing rice bran as its substrate. The increased demand for this fungus has not been met with successful cultivation methods for fruiting body production in natural environments. Therefore, we tested the effect on the growth of P. tenuipes using a Solid media based on pests. In this results, the Solid media based on M.alternatus was effective in increasing the growth of P. tenuipes and the content of cordycepin. Moreover, we confirmed the conditions for manufacturing a Solid media based on M.alternatus for P. tenuipes growth. We suggested that the growth-promoting compounds offers valuable insights for optimizing fungal cultivation conditions, thereby enhancing productivity and contributing to a broader understanding of fungal physiology in varying nutritional environments.

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.

In this study, the inhibitory effect of Paecilomyces tenuipes extract on PSA and angiogenesis-related factor expression levels were investigated in human prostate cancer cells, LNCaP. P. tenuipes extract significantly inhibited PSA expression in a dose-dependent manner. We also investigated the inhibitory effect of P. tenuipes extract on the expression of angiogenesis-related genes including VEGF, MMP-2, MMP-9, TIMP-1, and TIMP-2. P. tenuipes extract significantly down-regulated the expression of MMP-2 and MMP-9 in a dose-dependent manner. On the contrary, P. tenuipes increased the expression of TIMP-1 and TIMP-2. Our findings indicate that P. tenuipes exhibits an inhibitory effect on angiogenesis in human prostate cancer cells.

In order to develop fermented silkworm “Dongchunghacho” (Paecilomyces tenuipes) with improved absorption and increased effectiveness, we fermented Dongchunghacho using four kinds of microorganisms, viz., lactic acid bacteria, Bacillus subtilis, Natto bacillus, and yeast. A total of 15 samples were fermented using a combination of microbial inoculation culture and conditions to produce fermentation products. The contents of basic components such as sugar, reducing sugar, protein, total polyphenol, and total flavonoid were examined as well as the antioxidant, tyrosinase inhibitory, and thrombolytic activities of the fermented products were analyzed. We observed that reducing sugar and protein contents decreased in most of the fermented products, but the products fermented using yeast exhibited higher sugar content and, thus, higher sweetness. Total polyphenol content and antioxidant activity did not increase in fermented products compared to non-fermented Dongchunghachos, but total flavonoid content and tyrosinase inhibitory and thrombolytic activities increased by fermentation. In particular, total flavonoid content and tyrosinase inhibitory and thrombolytic activities primarily increased in the products fermented using yeast and lactic acid bacteria. However, it was not possible to confirm the increase in these activities in samples fermented by single fermentation using only yeast. Therefore, we propose that it will be possible to develop fermented food from silkworm Dongchunghacho (P. tenuipes) with excellent health benefits through additional study of multiple fermentation conditions using lactic acid bacteria and yeast.

To synnema formation and entomopathogenic of Paecilomyces tenuipes using some of media under the blue LED lamp. Lyophilized matured silkworm hemolymph and Bombyx mori powder were used as nutrient source in cultivation of an entomopathogenic fungus, Paecilomyces tenuipes. Matured silkworm (MS, aged seventh days of fifth instars) was in whole lyophilized without any treatment. Hemolymph (HL) powder was prepared by water-soluble extraction from matured silkworms which was silk gland removed and also lyophilized, too. Both of media showed better growth rates and synnemata formations than potato dextrose (PD) medium, and B. mori was controlled under the blue LED lamp conditions. To investigate their effect on mycelial growth of P. tenuipes, each of them were provided as nutrient sources by final concentrations of 2, 3 and 4%, respectively. Total 18 test cultures were cultivated for 14 days with no light. Three PD cultures were used as control group. Each of growth areas was measured with an image analyzer. The result showed that MS and HL medium promoted mycelial growth of P. tenuipes by 287 and 310%, respectively, in comparison with PD medium. The synnema formation and its shape of Paecilomyces tenuipes using B. mori under the blue LED lamp group are not so good than normal conditions. But, how to change of a compositions are not yet. It will be investigation in the future.

The genomic structure and phylogenetic relationships of HSP88 genes from P. tenuipes Jocheon-1, P. tenuipes, C. militaris and C. pruinosa are described. The HSP88 genomic DNA from P. tenuipes Jocheon-1, P. tenuipes and C. militaris all contain 5 introns and 6 exons with the length of 13, 62, 32, 1438, 306, 288 bp, encoding 713 amino acid residues. C. pruinosa HSP88 genomic DNA contains 4 introns and 5 exons encoding 713 amino acids. The length of each exon of C. pruinosa HSP88 is 13, 62, 32, 1744, 288 bp and the length of exon 4 is identical to the total length of exon 4 and exon 5 of HSP88 of P. tenuipes Jochoen-1, P. tenuipes, and C. militaris. The deduced amino acid sequence of P. tenuipes Jocheon-1 HSP88 showed 99% identity with the P. tenuipes, 97% identity with the Cordyceps militaris, and 98% identity with the C. pruinosa. Phylogenetic analysis confirmed that the P. tenuipes Jocheon-1, P. tenuipes, C. militaris and C. pruinosa HSP88 are placed together within the ascomycetes group of fungal clade.

In this study, a full-length heat shock protein88 complementary DNA (cDNA) of Paecilomyces tenuipes Jocheon-1 was obtained by screening of P. tenuipesJocheon-1 Uni-Zap cDNA library and 5' RACE polymerase chain reaction. The Paecilomyces tenuipes Jocheon-1 heat shock protein88 cDNA contains an open reading frame of 2,139 bp encoding 713 amino acid residues. The deduced amino acid sequence of the P. tenuipes Jocheon-1 HSP88 cDNA showed 77% identity to N. haematococca HSP88 and 45-76% identity to other fungi HSP88. Phylogenetic analysis and BLAST program analysis confirmed that the deduced amino acid sequences of the P. tenuipes Jocheon-1 HSP88 gene belonged to the ascomycetes group within the fungal clade and P. tenuipes Jocheon-1 HSP88 also contains the conserved ATPase domain at the N-terminal. The cDNA encoding P. tenuipes Jocheon-1 HSP88 was expressed as a 88 kDa polypeptide in baculovirus-infected insect Sf9 cells. Under different stress conditions, mRNA expression of P. tenuipes Jocheon-1 HSP88 were quantified by real-time PCR and the result showed that heat shock stress affected the mRNA expression levels of P. tenuipes Jocheon-1 HSP88.

Presently, We have constructed an olig-d(T) primed directional cDNA library from the silkworm Dongchunghacho, an entomopathogenic fungus, of which species is belonging to Paecilomyces tenuipes Jocheon-1. To isolate and screen genes in the fungus, 626 expressed sequence tags(ESTs) were generated by a partial sequencing from the cDNA library. Paecilomyces tenuipes Jocheon-1 cDNA encoding the glyceraldehyde-3-phosphate dehydrogenase(Pt-GAPDH) of Paecilomyces tenuipes Jocheon-1 was cloned from the above cDNA library. The complete cDNA sequence of Pt-GAPDHis comprised of 1,014bp encoding 338 amino acid residues. The deduced protein sequence of Pt-GAPDH showed higher homology with Beauberia bassiana-GAPDH(93% amino acid identity). Hydropathy analysis revealed that Pt-GAPDH protein is hydrophilic. The major three amino acids in its composition of amino acid residues were alanine(11.54%), valine(9.47%) and glycine(8.88%). The cDNA encoding Pt-GAPDH was expressed as a 37 kDa polypeptide in baculovirus-infected insect Sf9 cells. The Pt-GAPDH gene of Paecilomyces tenuipes entomopathogenic fungus consisted of three exons and two introns coding for 338 amino acid residues, and the genomic DNA length of the gene spans 1302bp. The accession number of the gene in GenBank are GU997099 for Pt-GAPDH cDNA and GU997102 for Pt-GAPDH genomic DNA.

Fungi belonging to the Paecilomyces spp. have recently been used as food and herbal medicines in Korea and are greatly popular as commercially available powdered supplement or dried fruiting body. Despite this acceptance and its use, little is known of the genes related to its reactive agents. Presently, We have constructed an olig-d(T) primed directional cDNA library from the silkworm Dongchunghacho, an entomopathogenic fungus, of which species is belonging to Paecilomyces spp. based on the previous identification of ITS1 and ITS2 at the molecular level and collected from Jocheon Miryang, Korea. To isolate and screen genes in the fungus, 626 expressed sequence tags(ESTs) were generated by a partial sequencing from the cDNA library. cDNA encoding the glyceraldehyde-3-phosphate dehydrogenase(Pt-GAPDH) of Paecilomyces tenuipes- Jocheon was cloned from the above cDNA library. The complete cDNA sequence of Pt-GAPDH is comprised of 1,014bp encoding 338 amino acid residues. The deduced protein sequence of Pt-GAPDH showed higher homology with Beauberia bassiana-GAPDH(93% amino acid identity). Hydropathy analysis revealed that Pt-GAPDH protein is hydrophilic. The major three amino acids in its composition of amino acid residues were alanine(11.54%), valine(9.47%) and glycine(8.88%). The Pt-GAPDH gene of Paecilomyces tenuipes entomopathogenic fungus consisted of three exons and two introns coding for 338 amino acid residues, and the genomic DNA length of the gene spans 1302bp. The accession number of the gene in GenBank are GU997099 for Pt-GAPDH cDNA and GU997102 for Pt-GAPDH genomic DNA. More investigation works including gene expression, immunological analysis etc. will be carried continuously without hesitation after this presentation.

The cultural characteristics and antibacterial activities of Cordyceps militaris and Paecilomyces tenuipes were compared. The mycelial growth was the highest on MCM (Mushroom Complete Medium) for C. militaris and on YMA (Yeast Malt Agar) for P. tenuipes. But the mycelial density on MMM (Mushroom Minimal Medium) was lower than other on media. The optimum mycelial growth was observed at 25℃. C. militaris was low mycelial growth when it was transferred over 5 times generation. The carbon source for the optimum mycelial growth was fructose of monosaccharide, maltose of disaccharide and dextrin of polysaccharide. The calcium nitrate of organonitrogen was found the best mycelial growth on C. militaris, while the sodium nitrate observed to be well for mycelial growth on P. tenuipes. The ammonium tartrate was observed to be the best among the inorganonitrogen used for mycelial growth. Antibacterial activities were found out just C. militaris against Bacillus cereus of Gram (+).

The growth characteristics of Paecilomyces tenuipes according to the passage in the two kind of liquid media were investigated by comparing the mycelium and conidium formation degrees. The potato dextrose broth medium and the silkworm larvae medium containing the silkworm powder were used as the liquid media, and the potato dextrose agar medium and the brown rice medium containing the powder of silkworm pupa were used as the solid media. The conidium formation degree in liquid media differed by the passages but that in solid media was not. This suggested that the passage in liquid media did not affect significantly the conidium formation in solid media. When the brown rice media were inoculated with the concentration of 1×10¹⁰ conidia/㎖, 1×10⁸ conidia/㎖ and 1×10⁶ conidia/㎖, respectively, the conidium formation degrees were similar. This indicated that the optimal inoculation concentration of conidium to the brown rice media is 1×10⁶ conidia/㎖.

경제적 가치가 매우 높은 눈꽃동충하초를 누에 품종별로 생리활성에 관한 연구를 수행한 결과를 요약하면 다음과 같다. 1. 눈꽃동충하초의 유리당 함량은 백옥잠과 하초잠에서는 Glucose함량이 가장 높았으며, 칠보잠과 양원잠에서는 Mannitol 함량이 많았다. 2. 눈꽃동충하초에 함유된 아미노산의 총 평균 함량은 10.3 μmole/g이며, 백옥잠이 10.7 μmole/g로 가장 높았으며, 하초잠과 칠보잠이 10.5 μmole/g, 양원잠이 0.96μmole/g로 가장 낮은 함량을 나타내었다. 16종의 구성아미노산 분포비율은 Arginine이 12.2%로 가장 높았으며, 다음으로 Glycine(10.5%) ＞ Proline(9.6)＞Tyrosine(8.9%) 순이었으며, Methionine이 2.1%로 가장 낮게 분포하였다. 3. 눈꽃동충하초의 유리 지방산 조성은 누에 품종에 관계없이 Oleic acid 함량이 가장 높았으며, 다음이 Linolenic acid, Palmitic acid, Linoleic acid 순이었다.

눈꽃동충하초 분생자경속을 형성하는데 액체배양을 이용한 접종원의 배지는 PDB가 가장 우수하였으며, 분생자경속 형성에 유리한 배양온도는 20℃로 나타났다. 조명은 500lx.에서 가장 양호한 성장을 하였으며 광조사 시간은 18hr이 가장 유리한 분생자경형성 조건을 나타내었다. 알맞은 배지 조건은 번데기와 현미를 50 : 50 의 비율에 증류수를 현미에 1 : 1비율로 첨가한 혼합배지에서 높게 나타났다. 번데기배지는 원형상태와 분쇄상태 중에서는 분쇄된 상태에서 번데기배지의 공극률이 높아 균사의 침입이 용이하여 분생자경속 회수율과 균사의 배지내에 형성이 원형상태의 번데기보다 더욱 높게 나타났으며, 균사의 생장도 역시 분쇄한 번데기에서 더욱 빠르고 고르게 나타났다. 종균 접종량은 15㎖의 종균을 접종하면 분생자경속이 잘 형성되었다. 눈꽃동충하초 재배에 가장 안정하게 재배하려면 분쇄한 번데기에 현미와 혼합한 배지에 종균 15 ㎖를 접종하여 24 에서 발이를 시킨 후 20℃의 배양실에서 재배하는 것이 가장 좋은 것으로 나타났다.

본시험은 한국에서 채집되어 강원대학교 동충하초은행(EFCC)에서 보관 중인 눈꽃동충하초의 분포와 배양적 특성을 조사하였다. 눈꽃동충하초는 한국의 산이나 계곡에서 어렵지 않게 채집할 수 있으며 골고루 분포한다. 채집시기는 7∼9월로 나타났으며 9월말에 가장 많이 채집할 수 있으며 채집된 눈꽃동충하초의 종은 눈꽃동충하초(P. tenuipes)가 가장 높게 채집되었다. 눈꽃동충하초의 균사배양 최적배지는 PDA배지이며 SDAY와 HMA배지에서도 생장은 가능하며 균사생장에 가장 적합한 온도는 25℃이며 pH는 6.0∼7.0으로 나타났다. 적합한 영양원으로는 탄소원은 dextrose이었으며, 질소원은 tryptone가 가장 우수하였고, 무기염으로는 KH2PO4에서 가장 고른 성장을 보였다. C/N에서는 20 : 1의 비율이 가장 적합한 것으로 조사되었다.

본 연구에서는 야생에서 분리한 눈꽃동충하초균의 배양적 특성을 알아보기 위해 배양특성 시험을 수행하였다. , 습도 60%에서 눈꽃동충하초균을 고체배양한 결과 초기 균사의 직경 12 mm에서 4일차에 균사체 직경이 36 mm 이었으며, 배양 5일차부터 급격한 균사신장이 일어나다가 8일차 이후부터는 생육속도가 서서히 감소되는 경향을 보였다. PDB 액체배지에서 종균배양의 상태와 생육도를 검토한 결과, PMV가 배양일차에 따라 1일차에 1%, 3일차에