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).

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.

Beet armyworm, Spodoptera exigua, diamondback moth, Plutella xylostella and tobacco cutworm, Spodoptera litura are the three most serious pests of many economically important crops such as cruciferous crops, various vegetables and ornamental plants. Because these pests are known to be resistant to lots of chemical insecticides, integrated control using both or either entomophathogen and/or natural enemy is thought to be an attractive alternative for effective control. One of the obstacles using and expanding mycopesticide is narrow host ranges. At commercial farms, cultivating crops are seriously damaged by various Lepidopteran pests. Farmers want to use a microbial control agent which can control various host insects to reduce cost and labor. In previous study, we selected two entomopathogenic fungi, Metarhizium anisopliae and Paecilomyces fumosoroseus, which shown high virulence against beet armyworm. For wide use of the isolates in farm, we tested its host ranges, especially to diamondback moth and tobacco cutworm, which also are serious pest in Korea. The two isolates were shown a good control effect in leaf disc bioassy.

Microbe have been considered as potential control agents for pest, as alternative to chemical control methods. Among entomopathogens, fungi cause the mortality by penetrating the cuticle of pest and/or by metabolites such as toxin. Not only this direct control effect of fungi, but repellency of fungi also may be used to prevent the pest. Repellence effect of fungi is considered as inhibitory factor to control termite. A study was reported in Japan that termite was able to detect and remove the conidia of fungi on their surface. The termite can escape from fungal infection and protect their colony. There is few study that insect pest such as moth can detect and avoid the virulence fungi. Therefore, we has been conducting the detection and avoidance of beet armyworm to high pathogenic fungi, Paecilomyces fumosoroseus. Adult of the beet armyworm avoided oviposition at Chinese cabbage treated with P. fumosoroseus compare to control. This result may be used to prevent the infestation of moth in crop production.

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 two-spotted spider mite Tetranychus urticae is a worldwide crop pest with a high insecticide resistance and an extensive host range. The aim of the present study was to evaluate the effect of PaeciPora®, which was formulated from the aerial conidia of an entomopathogenic fungus Paecilomyces lilacinus strain HY-4, to control T. urticae in cucumber field. In the field study, conidia of P. lilacinus HY-4 and a chemical acaricide azocyclotin were investigated for their control of the adult females of T. urticae. The strain produced a mortality of 56.0% on day 3 and 63.6% on day 7 post-treatment respectively at 1×107 conidia/mL, and no evidence of a mortality benefit was seen in the control group. Additionally, in the pesticide injury test, no agrochemical damage was found in hot pepper, watermelon, Chinese cabbage, oriental melon or strawberry by spraying PaeciPora® on them. The results indicated the possibility of the use of P. lilacinus HY-4 as a microbiological control agent against T. urticae in the Integrated Pest Management program.

An entomopathogenic filamentous fungus, Paecilomyces lilacinus strain HY-4, has a great potential as a promising bio-pesticide due to its superior pathogenicity against Adoretus tenuimaculatus and Tetranychus urticae. When the fungal strain infects host cuticle, it secrets a combination of hydrolytic enzymes including chitinase to solubilize the cuticle. Thus, we investigated effects of different carbon and nitrogen sources on the production of a chitinase from P. lilacinus strain HY-4. The organism produced an extracellular chitinase at a relatively high level (45.4 mU/ml) when cultivated for 5 days on a medium supplemented with insect pupa (0.5%) and colloidal chitin (1%), which was prepared by treating chitin from crab shells (Sigma-Aldrich Co. Ltd.) with 12 N HCl solution. However, extracellular secretion of chitinase by strain HY-4 was found to be significantly repressed in the presence of glucose (1%).

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.

Paecilomyces lilacinus HY-4 is an entomopathogenic filamentous fungus that has exhibited insecticidal activity against Adoretus tenuimaculatus and Tetranychus urticae. Strain HY-4 has attracted a great deal of industrial concerns because this organism can be applicable as a potent bio-pesticide. In this study, we developed an optimal diphasic fermentation technique for HY-4 conidial production. The substrate prescription which was made up of hulls or brans was obtained by screening of agricultural products, and the conidial production could reach a minimum of 2.5×1010 conidia/g after solid fermentation for 12 days in a given condition. It was interesting to note that strain HY-4 was propitious to sporulate more efficiently and productively in liquid fermentation process in the presence of insect pupa. For the industrial application of the HY-4 spores, their pH and thermal stabilities were evaluated on SDA agar plate as well. Additionally, the germination rate of HY-4 spores was still above 90% even when they were stored for 5 months at ambient temperature.

This study was carried to investigate the antidiabetic and lipid metabolism of water extract paecilomyces japonica(PJ) in Streptozotocin (STZ) induced diabetic rats. Diabetes were induced by intravenous injection of STZ at a dose of 42mg/kg dissolved in citrate buffer. The water extract of paecilomyces japonica were orally administrated once a day for 7 days at a dose of 500mg/kg or 1,000mg/kg. The contents of serum glucose, triglyceride(TG), total cholesterol were significantly decreased in PJ treated group compared to the those of STZ-control group. The content of hepatic glycogen and activities of glucose-6-phosphate dehydrogenase(G-6-PDH), glucokinase(GK) were significantly increased, but activity of glucose-6-phoshatase(G-6-Pase) was significantly decreased in PJ treated group compared to the those of STZ-control group. These results indicated that water extract of paecilomyces japonica would have antidiabetic and lipid metabolism effect in STZ-induced diabetic rats.

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/㎖.

This study was carried out to investigate the growth inhibition of Propionibacterium acnes by mycelial culture broth of Paecilomyces japonica in the Mulberry leaf extract. The results are as following. The growth inhibition effect of P. japonica mycelial culture broth against P. acnes in various concentration of Mulberry leaf extract was the most effective in 3% Mulberry leaf extract. The inhibition effect of P. japonica mycelial culture broth against P. acnes in Mulberry leaf extract according to the culture time was the moust effective for 25 days cultivation. As the treating volume of the mycelial culture broth was increased, the growth inhibition effect against P. acnes was increased as well. The growth inhibition effect of mycelial culture broth against P. acnes according to the time of heat treatment was active by 45min at 100℃, while it was inactive at more than 60min. Taken together P. japonica mycelial culture in the Mulbarry leaf extract has a possibility to be an element of skincare cosmetics regulating the acnes. This means that the enzyme may be used for health-care such as thrombosis without any hamful behavour in the blood vessel.