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Beauveria bassiana is a common fungal pathogen of Protaetia brevitarsis larvae, and although it is less common than Metarhizium anisopliae, the pathogen still poses a great risk to humans and animals that consume infected insects, owing to B. bassiana`s production of toxins like beauvericin and mycotoxin. Interestingly, the beneficial microorganism Saccharomyces cerevisiae possesses antifungal properties. In the present study, we found that S. cerevisiae inhibited the growth of B. bassiana by 97% and that S. cerevisiae failed to harm P. brevitarsis when administered via intracoelomic injection (1 × 107 cfu/mL). In addition, we also found that S. cerevisiae consumption increased the survival time of percutaneously infected P. brevitarsis larvae by 5 d and reduced the mortality of infected larvae by 12%. Therefore, S. cerevisiae is expected to be useful in the prevention and control of B. bassiana in the production of P. brevitarsis larvae.

In a previous report, we identified several candidate antimicrobial peptides through de novo RNA sequencing of the centipede Scolopendra subspinipes mutilans . Here, we identify and characterize one of these peptides, Scolopendrasin I. We identified the centipede antimicrobial peptide Cecropin from the centipede transcriptome using an SVM algorithm, and subsequently analyzed the amino acid sequence for predicted secondary structure using a GOR algorithm. We identified an alpha helical region of Cecropin and named it Scolopendrasin I. We then assessed antimicrobial and hemolytic activity of Scolopendrasin I. Scolopendrasin I showed antimicrobial activity against various microbes, including antibiotic-resistant Gram-negative bacteria, in a radial diffusion assay. Scolopendrasin I had potent antibacterial activity against acne-associated microbes in a colony count assay and showed no hemolytic activity in a hemolysis assay. In addition, we confirmed that Scolopendrasin I bound to the surface of bacteria via a specific interaction with lipoteichoic acid and lipopolysaccharide, two components of bacterial cell membranes. In conclusion, the results presented here provide evidence that this is an efficient strategy for antimicrobial peptide candidate identification and that Scolopendrasin I has potential for successful antibiotic development.

CopA3 is a homodimeric α-helical peptide derived from coprisin which is a defensin-like antimicrobial peptide that was identified from the dung beetle, Copris tripartitus. CopA3 has been reported to have anticancer activity against leukemia cancer cells. In the present study, we investigated the anticancer activity of CopA3 in human gastric cancer cells. CopA3 reduced cell viability and it was cytotoxic to gastric cancer cells in the MTS and LDH release assay, respectively. CopA3 was shown to induce necrotic cell death of the gastric cancer cells by flow cytometric analysis and acridine orange/ethidium bromide staining. CopA3-induced cell death was mediated by specific interactions with phosphatidylserine, a membrane component of cancer cells. Taken together, these data indicated that CopA3 mainly caused necrosis of gastric cancer cells, probably through interactions with phosphatidylserine, which suggests the potential utility of CopA3 as a cancer therapeutic. [BMB Reports 2015; 48(6): 324-329]

In this study, the compositions of Ophiocordyceps longissima mycelia, synnemata, and fruiting bodies were first analyzed in order to clarify its chemical basis for development as a health food or medicine. We found that the contents of crude protein, polysaccharides, and macroelements were highest in mycelia, whereas effective components, including mannitol, ergosterol, adenosine, inosine, Zn, and Se, were lowest in mycelia. Polysaccharide, mannitol, and ergosterol levels in synnemata (2.33, 4.54, and 0.66 g/100 g, respectively) were similar to those in fruiting bodies, but was significantly different from those of mycelia (4.79, 1.77, and 0.43 g/100 g). Trehalose content in fruiting bodies (2.15 g/100 g) was >4 times higher than that in synnemata (0.5 g/100 g). Adenosine content in fruiting bodies (0.024 g/100 g) was 3-4 times higher than that of synnemata, whereas inosine, cytosine, guanosine, and uridine (0.093, 0.145, 0.053, and 0.073 g/100 g) were highest in synnemata. Cu and Se were lower in mycelia (3.30 × 10⁻⁴ and 3.1 × 10⁻⁵ g/100 g) than in synnemata (1.85 × 10⁻³ and 7.2 × 10⁻⁵ g/100 g) and fruiting bodies (1.67 × 10⁻³ and 4.3 × 10⁻⁵ g/100 g). As, Hg, Cd, and Pb in mycelia were under the limit of edible fungus health standard; Pb was not found, but Cd, Hg, and As were detected in fruiting bodies. These findings suggest that O. longissima could be utilized in different culture methods according to market demand and might be a possible health food or medicinal resource.

The purpose of this study was to determine the possibility of black soldier fly (Hermetia illucens) as a animal feedstuff.Insect resources have been widely recognized that it exhibits its own biological activity by whole body or its metabolicintermediates. To accomplish this, its general and biological activities were measured. The present study was conductedto compare antioxidant activity of black soldier fly larva, pupa and grasshopper by examining radical scavenging activ-ity using DPPH (2,2 diphenyl 1-picryl hydrazyl). DPPH radical scavenging activity was conducted on the EtOH,MeOH, H2O fractions of black soldier fly larvae. Results showed that black soldier fly pupa on the H2O fraction con-tained the highest DPPH radical scavenging activity among the samples. FRAP assay are more suitable methods to eval-uate antioxidant activity of black soldier fly extracts. Among the samples, the FRAP value showed higher antioxidantactivity in the extracts from black soldier fly pupa extract on H2O fraction. These results suggest that black soldier flylarva and pupa extract has antioxidant activity and its fractions can be utilized to develop functional feedstuff.Key words : Black soldier fly, Hermetia illucens, Antioxidant activity

This study was performed to determine the whitening effect of organic solvent extracts from the centipede, Scolopendrasubspinipes mutilans. We prepared different concentrations (50%, 70% and 100%) of ethanol, methanol, 100% ethylacetate and water extracts. We tested melanin inhibitory effect and tyrosinase activity using B16/F10 melanoma cell.As a result, treatment of organic solvent extracts is decreased the biosynthesis of melanin and tyrosinase activity to36 ~ 86%. Especially the 70% ethanol extracts was the most effective in B16/F10 melanoma cells. In the study on mel-anogenic protein expression, 70% ethanol extracts of Scolopendra subspinipes mutilans blocked glycosylation of tyro-sinase. Therefore this result suggests that 70% ethanol extracts could be developed as skin whitening agents.Key words : Scolopendra subspinipes mutilans, Glycosylation, Tyrosinase, Melanin, Organic solvent extracts 서론 피부가 자외선의 노출로 인해 자극을 받게 되면 멜라닌을 생성하여 자외선으로 야기되는 손상으로부터 피부를보호한다(Hwang et al. 2013). 하지만 오늘날 과도한 자외선의 노출로 인해 멜라닌(melanin)이 과다 생성되면서 기미, 주근깨 등의 피부 반점이 형성되고 더 나아가서는 피부 노화, 피부암 등을 야기시켜 피부 건강을 위협하고 있다(You et al. 2009). 따라서 최근 식물, 곰팡이 등 여러천연물 추출물들을 이용한 멜라닌 합성 저해를 위한 연구가 활발히 진행되고 있다(Oh et al. 2007).멜라닌은 적, 노랑색의 페오멜라닌(pheomelanin)과 흑,갈색의 유멜라닌(eumelanin)으로 구분되는데(Ko and Kim2010) 이는 티로시나아제(tyrosinase) 효소에 의해 조절되며(Jeon et al. 2009), 특히 유멜라닌의 경우 티로시나아제와 함께 TRP-1, TRP-2 효소에 의해 조절된다고 알려져있다(Kim et al. 2011). 멜라닌의 구체적인 합성경로는 멜라닌 형성세포내의 멜라노좀(melanosome)에 의해 만들어지며(Sim et al. 2008) 형성된 멜라닌은 피부에 축적되어색소 침착 현상이 일어나게 되는데 이는 L-티로신(tyrosine)을 기질로 티로시나아제에 의해 L-DOPA, DOPA quinine,DOPA chrome을 거쳐 최종 단계로 멜라닌 polymer를 형성한다(Kim et al. 2010). 따라서 멜라닌 합성 저해제의개발은 티로시나아제의 활성 억제가 중요환 부분을 차지하고 있으며 지금까지 알려진 티로시나아제 억제제로는 알부틴(albutin), kojic acid 등이 있다(Sim et al. 2008). 또한fatty acid, glucosamine 등은 티로시나아제의 glycosylation을 저해하여 멜라닌의 합성을 저해하는 저해제로 알려져있다(Jeon et al. 2009). 왕지네(Scolopendra subspinipes mutilans)는 왕지네목 왕지네과의 절지동물로 한국, 중국, 일본 등지에 많이 분포하며, 몸 빛깔은 흑록색이고 머리는 붉은색이며, 몸은 수

The centipede Scolopendra subpinipes mutilans is a medicinally important arthropod species. However, its transcriptome is not currently available and transcriptome analysis would be useful in providing insight into a molecular level approach. Hence, we performed de novo RNA sequencing of S. subpinipes mutilans using next-generation sequencing. We generated a novel peptide (scolopendrasin II) based on a SVM algorithm, and biochemically evaluated the in vitro antimicrobial activity of scolopendrasin II against various microbes. Scolopendrasin II showed antibacterial activities against gram-positive and -negative bacterial strains, including the yeast Candida albicans and antibiotic-resistant gram-negative bacteria, as determined by a radial diffusion assay and colony count assay without hemolytic activity. In addition, we confirmed that scolopendrasin II bound to the surface of bacteria through a specific interaction with lipoteichoic acid and a lipopolysaccharide, which was one of the bacterial cell-wall components. In conclusion, our results suggest that scolopendrasin II may be useful for developing peptide antibiotics.

This experiment was conducted to investigate the effect of dietary supplementation of the black soldier fly (bsf) larvaeon growth performance, carcass rate, and fatty acid profile of chicken meat in broiler chicks. Four hundred eighty 1-d-old broilers (Ross 308) were distributed into 3 treatments with 4 pens at 30 birds per replicate. Treatments were corn-soybean meal based broiler diet (control) and the basal diet replaced with bsf at the level of 3 and 6% respectively. Via-bility was not different significantly. Body weight gain was higher in control than other treatments. Feed intake was notstatistically different of treatments. Feed conversion ratio was lower in control than other treatments. Relative weightof breast meat was significantly increased in all treated groups than in control. But, another relative weights of chickenmeat cuts (thigh, wing, neck, back) were not significantly different among the groups. In conclusion, it could be sug-gested that dietary supplementation of black soldier flylarvae could beneficially improved the performanceon broiler.Key words : black soldier fly (bsf), Hermetia illucens, organic wastes

Many studies have explored suppression of aflatoxin produced by Aspergillus Genus. On the other hand, this study examined the inhibitory effect of the culture broth extract (CE) of A. tamarii obtained from dead silkworm on nitric oxide (NO) production and its antioxidative activity. The culture broth was extracted with EtOAc, dried, and then used in this experiment. As a result, CE did not show cytotoxicity on RAW 264.7 cells at any concentration. Moreover, CE suppressed lipopolysaccharide (LPS)-induced NO production of RAW 264.7 cells in a dose-dependent manner. The total phenol content according to the Folin-Dennis method, the antioxidative activity by DPPH, and the nitrate radical scavengingcapacity of CE were increased in a dose-dependent manner. Thus, many of the phenolic compounds were considered to represent the antioxidative activity.

In this study, silk gland powder of silkworm were investigated to see the possibility for baby powder cosmetics materials. To test possibility as a baby powder cosmetics, total content rate of amino acids, DPPH free radical scavenging assay, MTT assay, and clinical trial were done. According to the result of the analysis of the amino acids of silk gland powder, serin (26.77%) content was the highest and asparatic acid (15.47%), and glycine (9.62%) were followed. DPPH free radical scavenging activity of silk gland powder was lower than vitamin C by 82.3% and 97%, respectively, which is relatively good. Moisture effect were increased in silk gland powder compared to control cosmetics by 50%. Also, silk gland powder was classified as a practically non-irritating material based on the score 0.05 of primary irritation index. Thus, these results suggest that silk gland powder of silkworm may have beneficial properties as a material for baby powder cosmetics.

Our previous study demonstrated that CopA3, a disulfide dimer of the coprisin peptide analogue (LLCIALRKK), has antibacterial activity. In this study, we assessed whether CopA3 caused cellular toxicity in various mammalian cell lines. CopA3 selectively caused a marked decrease in cell viability in Jurkat T, U937, and AML-2 cells (human leukemia cells), but was not cytotoxic to Caki or Hela cells. Fragmentation of DNA, a marker of apoptosis, was also confirmed in the leukemia cell lines, but not in the other cells. CopA3-induced apoptosis in leukemia cells was mediated by apoptosis inducing factor (AIF), indicating induction of a caspase-independent signaling pathway. [BMB reports 2012; 45(2): 85-90]

Cordyceps (vegetable wasp and plant worm), an entomopathogenic fungi, has been used as a herbal medicine in Asian countries since ancient times. Cordyceps nutans is common but there is little research on this species. This study investigated the optimal culture conditions of C. nutans and the inhibitory effect on nitric oxide (NO) production in RAW 264.7 cell treated culture broth. The optimal conditions for the mycelial growth were and pH 7.0-8.0. Mycelial growth was highest on mushroom complete medium (MCM), V8 juice agar (V8A), and yeast malt dextrose (YMD) medium. Mycelial growth on mushroom minimal medium (MMM) did not occur, so nutrient source was essential. Dextrose and sucrose as carbon sources, and ammonium citrate as a nitrogen source were satisfactory for mycelial growth. Cytotoxicity of C. nutans culture broth was not found in RAW 264.7 cells. C. nutans culture broth suppressed NO production of lipopolysaccharide (LPS)-stimulated RAW 264.7 cell in a dose-dependent manner. Thus, our results provided the optimal conditions for cultivation of C. nutans and showed that C. nutans may have excellent physiological activities.

Silk fibroin, a natural protein produced by silkworm, is a good biomaterial which has biodegradability and biocompatibility. To ascertain the effects of silk fibroin on cell growth, silk fibroin films were prepared using silk fibroin aqueous solutions of various concentrations. We investigated the attachment, proliferation, morphology of the cells and the expression levels of genes related to cell attachment and growth on the silk fibroin films. When the cells were cultured on the 0.1 and 1% silk fibroin film, the cell adhesion ability was very excellent. Particularly, overall cell growth on the 1% silk fibroin film was definitely superior to the others. Also, expression levels of genes related cell growth were increased on the 0.1 and 1% silk fibroin film. These results suggest silk as a material for medical applications.

Ovalicin, a secondary metabolite produced by an entomopathogenic fungus Metarhizium anisopliae var. anisopliae, is currently used as an efficient biological control agent against various agricultural insect pests, but not so many biological activities have been investigated. To assess whether ovalicin has potential in ameliorating atopic dermatitis-related symptoms in mice, we first sensitized skin in the dorsal neck of Balb/c mice using compound 48/80, and scrutinized whether the compound affected the atopic dermatitis-related symptoms. The results revealed that ovalicin significantly reduces scratching behavior in a concentration-dependent fashion. Moreover, the treatment inhibits the levels of the degranulation of mast cells by 65%, and levels of histamine release by 51% at a concentration of 10 µg/ml. Together, the present data strongly suggest that ovalicin elicits potential anti-atopic activities in mice.

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 . 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 caterpillar-shaped Chinese medicinal mushroom (DongChongXiaCao) looks like a worm in the winter and like a grass in the summer. The fruiting body has been regarded as popular folk or effective medicines used to treat human diseases such as asthma, bronchial and lung inflammation, and kidney disease. The fruiting bodies of Paecilomyces tenuipes that formed on the living silkworm (Bombyx mori) host were used in this examination. This study was carried out to investigate the proximate composition, soluble sugar, amino acid and fatty acid profiles, and contents of the bioactive ingredient including adenosine and D-mannitol in the fruiting-bodies. The moisture content was 57.56%. Soluble sugars found were glycerol, glucose, mannitol and sucrose, and the contents exceeded 24 mgg(-1)dry weight. Total free amino acid content was 17.09 mg g(-1)dry weight. Arginine, glycine, proline and tyrosine were main amino acids. The content of oleic acid in fatty acids was high. Adenosine was more abundant in fruiting bodies than corpus.

The effects of medium components and environmental factors on the production of mycelial biomass and polysaccharide–peptide complexes (exobiopolymers) by Cordyceps sphecocephala J-201 were investigated in submerged cultures. The optimal temperature and initial pH for the production of both mycelial biomass and exobiopolymers in flask cultures were found to be 25 °C and pH 4–5, respectively. The optimal combination of the media constituents was as follows (g l−1): sucrose 40, yeast extract 6, polypepton 2, KH2PO4 0.46, K2HPO4 1, and MgSO4·7H2O 0.5. The results of bioreactor culture revealed that the maximum concentration of mycelial biomass (28.2 g l−1) was obtained at an agitation speed of 300 rpm and at an aeration rate of 2 vvm, whereas maximum exobiopolymer production (2.5 g l−1) was achieved at a milder agitation speed (150 rpm). There was a significant variance in mycelial morphology between different aeration conditions. Looser mycelial pellets were developed, and their size and hairiness increased as the aeration rate increased from 0.5 to 2.0 vvm, resulting in enhanced exobiopolymer production. The apparent viscosities of fermentation broth increased rapidly towards the end of fermentations at the conditions of high aeration rate and agitation speed, which were mainly due to high amount of mycelial biomass rather than exobiopolymers at the later stages of fermentation. The three different exobiopolymers (FR-I, -II, and -III) were fractionated by a gel filtration chromatography on Sepharose CL-6B. The carbohydrate and protein contents in each fraction were significantly different and the molecular weights of FR-I, FR-II, and FR-III were determined to be 1831, 27, and 2.2 kDa, respectively. The compositional analysis revealed that the three fractions of crude exobiopolymers consisted of acidic and nonpolar amino acids, such as aspartic acid, glutamic acid, glycine, and valine in protein moiety, and of mainly mannose and galactose in sugar moiety.

The CopA3 dimer peptide is a coprisin analog that has an anticancer effect against human cancer cells in vitro. In this study, we investigated the anticancer activity of the enantiomeric CopA3 dimer peptide in human gastric cancer cell lines as well as in an in vivo tumor xenograft model. Enantiomeric CopA3 reduced gastric cancer cell viability and exhibited cytotoxicity against cancer cells. Enantiomeric CopA3-induced cell death was mediated by specific interactions with phosphatidylserine and phosphatidylcholine, membrane components that are enriched in cancer cells, in a calcein leakage assay. Moreover, acridine orange/ethidium bromide staining, flow cytometric analysis, and Western blot analysis showed that enantiomeric CopA3 induced apoptotic and necrotic gastric cancer cell death. The antitumor effect was also observed in a mouse tumor xenograft model in which intratumoral inoculation of the peptide resulted in a significant decrease in the SNU-668 gastric cancer tumor volume. In addition, periodic acid-Schiff and hematoxylin staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay revealed apoptotic and necrotic cell death in tumor masses treated with greater than 150 μg CopA3. Collectively, these results indicate that the enantiomeric CopA3 dimer peptide induces apoptosis and necrosis of gastric cancer cells in vitro and in vivo, indicating that the peptide is a potential candidate for the treatment of gastric cancer, which is a common cause of cancer and cancer deaths worldwide.

Previously, we performed de novo RNA sequencing of Scolopendra subspinipes mutilans using high-throughput sequencing technology and identified several antimicrobial peptide candidates. Among them, a cationic antimicrobial peptide, scolopendrasin VII, was selected based on its physicochemical properties, such as length, charge, and isoelectric point. Here, we assessed the anticancer activities of scolopendrasin VII against U937 and Jurkat leukemia cell lines. The results showed that scolopendrasin VII decreased the viability of the leukemia cells in MTS assays. Furthermore, flow cytometric analysis and acridine orange/ethidium bromide staining revealed that scolopendrasin VII induced necrosis in the leukemia cells. Scolopendrasin VII-induced necrosis was mediated by specific interaction with phosphatidylserine, which is enriched in the membrane of cancer cells. Taken together, these data indicated that scolopendrasin VII induced necrotic cell death in leukemia cells, probably through interaction with phosphatidylserine. The results provide a useful anticancer peptide candidate and an efficient strategy for new anticancer peptide development.

We compared the mRNA expression profile of the Harmonia axyridis larvae that were either untreated or treated with LPS. The extracted mRNAs were subjected to ACP RTPCR analysis using a combination of arbitrary primers and oligo (dT) primer. Among the 47 DEGs differentially expressed, we identified a cDNA showing homology with defensin-like antibacterial peptide. The cDNA showed a putative 32-residue signal sequence and a 50-residue mature peptide named harmoniasin. We also investigated the antibacterial activity of the harmoniasin analog, which exhibited potent antibacterial activities against Gramnegative and -positive bacteria strains and it also evidenced no hemolytic activity.

During search for novel bioactive materials from natural resources with the potential as health food and alternative medicine, the culture broth of Cordyceps longissima (CL) J106, J144 and C. scarabaeicola (CS) J94, J123 were prepared, and their effect on cytotoxicity and nitric oxide (NO) production in RAW 264.7 cells were investigated. Whereas the culture broth of CL J144 and CS J123 had cytotoxicity in RAW 264.7 cells, that of CL J106 and CS J94 did not. The culture broth of CL J106 and CS J94 suppressed NO production in RAW 264.7 cells activated with lipopolysaccharide (LPS) at a dose-dependent manner. These results suggest that culture broth, a by-product of Cordyceps, may have active compounds with anti-inflammatory effect. In addition, it appears that their biological activity is dependent on the strains in spite of the same species.

Beauveria bassiana is a fungi that is well-known for demonstrating a resistance to environmental change. To confirm whether S-(-)-10,11-dihydroxyfarnesic acid methyl ester (DHFAME) produced by Beauveria bassiana KACC46831 causes phototoxicity when used for cosmetic purposes due to its anti-tyrosinase activity, we conducted in vitro and in vivo phototoxicity tests. There were no significant changes or damage observed in the compound-treated group with regards to skin phototoxicity, while 8-methoxypsoralen, which served as a positive control, induced toxic effects. The in vitro 3T3 neutral red uptake assay, an alternative assessment, was used for further confirmation of the phototoxicity. The results showed that DHFAME did not exhibit phototoxicity at the designated concentrations, with or without UV irradiation in the 3T3 cells. These results indicated that the methyl ester produced by Beauveria bassiana KACC46831 does not induce phototoxicity in the skin. Therefore, the results of the present study indicate that DHFAME shows potential for use as a cosmetic ingredient that does not cause skin phototoxicity.

Harmoniasin is a defensin-like antimicrobial peptide identified from the ladybug Harmonia axyridis. Among the synthetic homodimer peptide analogues derived from harmoniasin, HaA4 has been found to have antibacterial activity without hemolytic activity. In this study, we investigated whether HaA4 has anticancer activity against human leukemia cell lines such as U937 and Jurkat cells. HaA4 manifested cytotoxicity and decreased the cell viability of U937 and Jurkat cells in MTS assay and LDH release assay. We found that HaA4 induced apoptotic and necrotic cell death of the leukemia cells using flow cytometric analysis, acridine orange/ethidium bromide staining and nucleosomal fragmentation of genomic DNA. Activation of caspase-7 and -9 and fragmentation of poly (ADP-ribose) polymerase was detected in the HaA4-treated leukemia cells, suggesting induction of a caspase-dependent apoptosis pathway by HaA4. Caspase-dependent apoptosis was further confirmed by reversal of the HaA4-induced viability reduction by treatment of Z-VAD-FMK, a pan-caspase inhibitor. In conclusion, HaA4 caused necrosis and caspase-dependent apoptosis in both U937 and Jurkat leukemia cells, which suggests potential utility of HaA4 as a cancer therapeutic agent.

Paecilomyces tenuipes reportedly have anticancer and immune activities, along with various other medicinal uses. Cultured products with P. tenuipes are certified for use in food in South Korea, and processed goods containing this fungus have been developed in many countries, particularly South Korea, Japan, and China. Research on mass production technology-procured raw materials for the manufacture of P. tenuipes is very important; however, cultures of the fungus have been unstable. This study identified stable cultivation conditions, focusing on growth inhibition and revitalization. Moisture regulation and preservation of pupae inoculated with P. tenuipes were used to control growth inhibition and revitalization. When inoculated silkworm pupae were dehydrated to 4% moisture and preserved freeze-dried or at -70 degrees C, -20 degrees C, or 4 degrees C, the mycelia in their bodies were able to survive for 14 d. Inoculated silkworm pupae were rehydrated for 3 h and the mycelia within their bodies were recovered at 94.3-96.3%. Silkworm pupae at 4% moisture were able to survive for 135 d at temperatures < 4 degrees C and for 1 y after freeze-drying. Optimal conditions for synnemata induction were 25 degrees C and 100-300 1x.

We report the isolation and characterization of a novel knottin-type antimicrobial peptide from the yellow-spotted long-horned beetle Psacothea hilaris. A cDNA encoding a 56-mer knottin-type propeptide was identified and its predicted molecular mass and pI were 5.92 kDa and 8.28, respectively. A 34-mer mature peptide was also selected and named herein as psacotheasin. The antimicrobial activity of chemically synthesized psacotheasin against human bacterial pathogens was subsequently investigated. The results showed that psacotheasin exerted potent activities against both Gram-positive and Gram-negative bacterial strains. The present study suggests that psacotheasin can be applied to develop novel therapeutic agents.