• Title/Summary/Keyword: protein extraction

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Mycelial Growth Using the Natural Product and Angiotensin Converting Enzyme Inhibition Activity of Pleurotus eryngii (천연물을 이용한 큰느타리 균사배양 및 Angiotensin Converting Enzyme 저해활성)

  • Kang, Tae-Su;Jeong, Heon-Sang;Lee, Myong-Yul;Park, Hee-Joeng;Jho, Taek-Sang;Ji, Seung-Taek;Shin, Myung-Keun
    • The Korean Journal of Mycology
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    • v.31 no.3
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    • pp.175-180
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    • 2003
  • To develop the health/functional food materials, we investigated the cultural condition of mycelial growth on the solid state fermentation using the brown rise, Acanthopanax sp. and Artemisia sp., and also evaluated inhibitory activity of angiotensin converting enzyme (ACE) of hot water extracts from cultured media of Pleurotus eryngii. As the amount of Acanthopanax nnd Artemisia In the cultural media increased, the mycelial growth rate decreased. Especially, addition of Aeantopanax showed marked effect than Artemisia. Moisture contents in three kinds of cultured media were in the range of $10.9{\sim}12.0%$. Crude protein fat and crude fiber content were the highest value in cultured brown rice medium, whereas the mineral contents (Ca, K and P) were higher in the Acanthopanax supplemented (5%) medium than the other media, The extraction yield of the Artemisia supplemented (5%) medium was the highest value of 4.80%, and the pH of hot water extract from cultured brown rice medium showed the lowest value of 6.1. Lightness (L) values in three kinds of extracts from cultured media were in the range of $85.8{\sim}87.1$. Redness (a) value was the highest In the brown rice and Acanthopanax supplemented media, however cultured Artemisia supplemented medium showed the highest value in yellowness (b). In comparison of sugar components analyzed by the thin layer chromatography with three kinds of samples, two spots were detected to be glucose and maltose, respectively. The ACE inhibitory activity of hot water extract from the cultured Acanthopanax supplemented medium showed the highest value at the concentration of $0.2{\sim}1.0\;mg/ml$. These results suggest that the Pleurotus eryngii grew in natural media using brown rice and Acanthopanax can be supplemented to the brown rice medium to enhance its ACE inhibitory activity as health/functional food materials.

Physicochemical properties and antioxidant activities of Streptococcus thermophilus KCCM 3782 strain fermented Cordyceps militaris grown on Tenebrio molitor (갈색거저리 동충하초의 Streptococcus thermophilus KCCM 3782 유산균 발효에 따른 물리화학적 특성과 산화방지 활성 비교)

  • Gim, Do-Woo;Park, Jun-Ki;Kim, Hyun-Jin;Cho, In-Kyung;Cho, Ja-Yong;Kim, Yong-Doo;Huh, Chang-Ki
    • Food Science and Preservation
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    • v.25 no.1
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    • pp.136-144
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    • 2018
  • This study aimed to provide basic evidence regarding the development of materials by analyzing the physicochemical properties and antioxidant activities of Streptococcus thermophilus KCCM 3782 strain-fermented Cordyceps militaris grown on Tenebrio molitor. Among the solvents tested, DPPH radical scavenging activity was the highest in the hot water-extracted sample after 30 min of extraction. Moisture content decreased, whereas crude protein and fat content increased, after lactic acid bacteria-mediated fermentation. Sodium, magnesium, calcium, and zinc contents increased, with potassium levels attaining the maximum value, whereas free amino acid content decreased after the fermentation. Among Hunter's color values, a value increased to $66.7{\rightarrow}149.92$ after fermentation, whereas the L and b values decreased to $15.79{\rightarrow}-15.75$ and $54.45{\rightarrow}0.01$, respectively. Cordycepin content assay increased from 7.02 mg% to 8.66 mg%. The DPPH free radical scavenging activity of the fermented product was 91.92 EDA%, which was higher than that of the extract (84.69 EDA%). The ABTS free radical scavenging and superoxide dismutase (SOD) activities were also higher in the fermented products.

Physicochemical properties of dried Saururus chinensis and the antioxidative activities of water and 70% ethanol extracts (덖음온도를 달리하여 전저리한 삼백초 건조물의 이화학적 특성 및 물과 70% 에탄올 추출물의 항산화효과)

  • Kang, Myung-Hwa;An, Su-Mi;Kim, Do-Hee
    • Journal of Nutrition and Health
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    • v.52 no.4
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    • pp.399-407
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    • 2019
  • Purpose: This study was conducted to evaluate the physicochemical properties of different batches of Saururus chinensis using different roasting temperature that were dried at different using different roasting temperatures and their were determined the antioxidative activities of water and 70% ethanol extracts. Methods: Extracts were examined for the total phenolic acid content, the and flavonoids contents and the antioxidant properties, including DPPH radical scavenging activity, ABTs scavenging activity and, the reducing power. Results: Moisture content was significantly higher in the LSC and the crude ash content was significantly higher in the HSC. The crude protein content was higher in the LSC (although not significantly), and the crude fat and carbohydrate contents were higher in the HSC (although not significantly). The total phenolic content was lower in the samples extracted with water, but there was no significant difference. However, the extracts extracted with 70% ethanol at a high drying temperature were significantly higher. The low temperature and high drying temperature batches of Saururus chinensis were significantly higher in the samples extracted with 70% ethanol than those extracted with 70% ethanol. The total phenolic acid content, the total flavonoid content and the electron donating ability were highest in the ethanol extract of Saururuschinensis treated at a high temperature. However, the ABTs radical activity was highest in the water extracted, high-temperature treated Saururuschinensis. The 70% ethanol extract of high temperature roasted Saururuschinensis had the highest antioxidative activities of all the Saururuschinensis batches. Conclusion: The total phenolic acid contents, total flavonoid contents, electron donating activity and reducing power activity were highest in all the 70% ethanol extraction batches of the high-temperature treated samples.

Key Foods selection using data from the 7th Korea National Health and Nutrition Examination Survey (2016-2018) (제7기 국민건강영양조사 (2016-2018) 자료를 활용한 한국인의 주요 식품 (Key Foods) 선정에 관한 연구)

  • Lee, Jung-Sug;Shim, Jee-Seon;Kim, Ki Nam;Lee, Hyun Sook;Chang, Moon-Jeong;Kim, Hye-Young
    • Journal of Nutrition and Health
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    • v.54 no.1
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    • pp.10-22
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    • 2021
  • Purpose: Key Foods refers to foods that have a high contribution in the nutrient intake of individuals, and exert important effects on their health. This study was undertaken to identify Korean Key Foods, using data from the 7th Korea National Health and Nutrition Examination Survey (KNHNES). Methods: The data source for the extraction of Key Foods was the 24-hour dietary survey data obtained from the 7th KNHNES (2016-2018), and 21,271 subjects were evaluated. A total of 17 nutrients were selected as the key nutrients for identifying the Key Foods, including energy, carbohydrates, protein, lipid, dietary fiber, calcium, phosphorus, iron, sodium, potassium, vitamin A, thiamin, riboflavin, niacin, vitamin C, cholesterol, and sugars. The nutrient consumption approach was applied to generate a list of potential Key Foods. Foods included in 85% of the cumulative intake contribution from one or more key nutrients, were subsequently selected as Key Foods. Results: Of the 1,728 foods consumed by survey respondents, we extracted 728 Key Foods. These Key Foods explained 94% key nutrient intakes of the subjects. Based on the contribution rate to key nutrient intake, the top 10 Key Foods identified were multigrain rice (5.32%), plain white rice (4.23%), milk (3.3%), cabbage kimchi (2.82%), grilled pork belly (1.56%), apples (1.52%), fried eggs (1.49%), cereal (1.36%), instant coffee mix (1.21%), and sweet potatoes (1.12%). These 10 foods accounted for 23.93% total key nutrient intake of the survey respondents. Conclusion: Seven hundred and twenty-eight foods were extracted and identified as the 2020 Korean Key Foods. These Key Foods can be considered the priority foods to be analyzed for establishing a national nutrient database.

Conservation and Scientific Analysis of Human Bone Excavated in Sabi Period of Baekje from Eungpyeong-ri, Buyeo (부여 응평리 출토 백제 사비기 인골 보존처리 및 과학적 분석)

  • KIM, Mijeong;LEE, Yunseop;CHO, Eunmin;PARK, Sujin;MOON, Minseong
    • Korean Journal of Heritage: History & Science
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    • v.55 no.1
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    • pp.305-321
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    • 2022
  • The stone chamber tomb in Eungpyeong-ri, Buyeo, is a joint tomb that contains the bodies of two individuals. This paper investigates the relationship between the buried persons and the characteristics of the stone chamber tomb. Based on the geographical location, relics, and the excavated human bones, it was determined that the tomb was built during the Sabi Period of the Baekje Dynasty and that the buried individuals were most probably residents of high stature or government officials. To study the excavated bones, the remains were carefully collected and conservation was carried out. Before collecting samples from the human bones for the analytical research, the results of near-infrared analysis were used to collect the samples for the isotope analysis and DNA analysis. The most important issue when handling the excavation site was the reinforcing agent and the concentration of the agent used. In situations like this, Paraloid B-72 is the most suitable agent. When the shape of human bones was difficult to distinguish from the soil, conservation was performed using X-ray and CT imaging data. The same chemical used for the reinforcement of the site was used to complete a minimum level of conservation to the surface areas where the conservation treatment of removing foreign substances, the reinforcement areas, and bonded areas were carried out. The collagen yield from the sample obtained at selected position was 3.8% to 6.1%. The results of analyzing the stable isotopes of carbon and nitrogen found in the extracted collagen showed that the stable isotope ratios came out to δ13C -18.3‰±0.1‰, -19.0‰±0.1‰ for EBW and δ15N 10.7‰±0.5‰, 10.6‰±0.1‰ for EBE. It is believed the two individuals consumed small amounts of minor cereals, mainly from C3 plants, and protein was obtained from eating terrestrial animals. What's more, the deviations in data obtained from the two individuals were so small that it could be inferred that the individuals ate similar foods. Considering the preservation state of the sample, amplifying DNA for the DNA analysis would have been very difficult since the amount of surviving DNA was so deficient. For DNA analysis, it is anticipated that the results could be derived by applying improved extraction methods that will be developed in the future. In this research, any association between scientific analysis(DNA and stable isotope ratio) and near-infrared spectroscopy was difficult to establish. Further research is needed on the utilization of near-infrared analysis for gathering samples from human bones.

Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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