It is crucial to develop a miniaturized cultivation method for large and rapid screening of high-yielding mutants of monacolin-K, a powerful anti-hypercholesterolemic secondary metabolite biosynthesized by the fungal cells of Monascus ruber. In order to investigate as many strains as possible in a short time, a miniaturized fermentation method especially suitable for the cultivation of the filamentous Monascus mutants was developed using $50m{\ell}$ culture-tube ($7m{\ell}$ of working volume) instead of the traditional $250m{\ell}$ flask ($50m{\ell}$ of working volume). Generally, in filamentous fungal cell fermentations, morphologies in growth and production cultures should be maintained as thick filamentous and compact-pelleted (usually less than 1 mm in diameter) forms, respectively, for enhanced production of secondary metabolites in final production cultures. In this study, we intended to induce the respective optimal morphologies in the miniaturized culture system for the purpose of rapid screening of overproducers. Miniaturized growth culture system was successfully developed due to the mass production of spores in the statistically optimized solid medium. When large amounts of spores were inoculated into the growth cultures, and brown rice flour (20 g/L) was also supplemented to the growth medium, dense filamentous morphologies were successfully induced in the growth cultures performed with the 50 ml culture tubes. It was implied that the amounts of spores inoculated into the growth tube-cultures and the growth medium components should be the key factors for the induction of the filamentous forms in the growth fermentations. Furthermore, in order to statistically optimize production medium, multiple experiments based on Plackett-Burman design and response surface method (RSM) were carried out, resulting in more than 2 fold enhanced production of monacolin-K in the final production cultures with the optimized production medium. Notably, under the production culture conditions with the statistically optimized medium, optimal pellet sizes below 1 mm in diameter were reproducibly induced, in contrast to the thick and viscous filamentous morphologies observed in the previous production cultures.
Lee, Ka Soon;Lee, Bo Hee;Seong, Bong Jae;Kim, Sun Ick;Han, Seung Ho;Kim, Gwan Hou;Park, Saet Byeol;Kim, Hyun Ho;Choi, Taek Yong
Journal of the Korean Society of Food Science and Nutrition
/
v.45
no.6
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pp.851-858
/
2016
The effective components of Schisandra chinensis are lignans (schizandrins and gomisins), which have various physiological functionalities such as anti-cancer, anti-inflammatory, and antioxidant activities. This study was carried out to determine the different parts of fruits in Schisandra chinensis to elevate their usefulness. Schisandra chinensis was separated into skin (epicarp), pulp (mesocarp), and seeds, and raw Omija and hot-dried Omija (HDO) were used as control. The most abundant component was nitrogen free extract (6.88~56.70%) followed by crude lipids (1.65~19.04%). The main mineral was K (383.10~2,024.10 mg/100 g), except in seeds where P was the main mineral. The main lignan in all parts of fruit was schizandrin, and the highest content of schizandrin was 9.46 mg/g in dried seeds. Total lignan content was 25.97 mg/g and 14.97 mg/g in dried seeds and HDO, respectively. A total of 17 components of fatty acids in seeds and HDO were detected, of which linoleic acid (72.66~73.78%), oleic acid (14.78~17.39%), palmitic acid (2.88~3.54%), and capric acid (1.70~4.93%) were determined as the major components. Main lignans and fatty acids of Schisandra chinensis fruit contain mainly seeds. Therefore, it is more efficient to use seeds than pulp and extract of fruit itself to use the components of Omija.
This study was conducted to compare the regeneration of the seedlings under different vegetation types and to identify the presence of allelopathy in Abies koreana(Ak) natural forest in Banyabong Peak(elevation, 1715m), of Mt. Chiri. Twenty quadrats($10m{\times}10m$) were placed in May, 1996 to classify vegetation structure using TWINSPAN. Water-soluble extracts from leaves and soil humus of different vegetation types were collected to test their effects on both seed germination of Ak and mycelial growth of ectomycorrhizal fungi. Phenolic compounds from soil humus were quantified using HPLC. Among the four vegetation types, Sasa borealis(Sb) was found in both Ak-Quercus mongolica(Qm) and Ak-Rhododendron schlippenbachii(Rs) communities. Natural seeding of Ak was $230,000{\pm}90,000seeds/ha$ in 1995 and their germination rate was 25% in an ideal laboratory condition. Density of Ak seedlings less than 5cm in height was 52,000/ha in 1996, while that of seedlings taller than 5cm in height was only 4,000/ha. In the case of Ak-Qm community, density of Ak seedlings with Sb understory was only 7% of the density of seedlings with Rs understary, suggesting the inhibitory effect of Sb. The germination rate of Ak seeds was significantly reduced by leaf extracts of Sb, and Rhododendron mucronulatum var. ciliatum(Rm) and Ak. Soil humus extract of Ak-Qm-Sb subcommunity reduced germination of Ak seeds by 81% and also reduced by 19% the respiration of mycelia of ectomycorrhizal fumgus, Lactarius sp. Among the seven phenolic compounds identified from the soil humus, extract, Ak-Qm-Sb subcommunity contained significantly high content of p-hydroxybenzoic acid, vanillic acid, and syringic acid. Particularly, p-hydroxybenzoic acid was present at 4.2ppm in the Sb roots and at 16.5ppm in the Sb humus, suggesting that it could be the primary allelopathic compound in Abies koreana forests with Sasa borealis understory.
The effects of seeding and clipping termination dates, and seeding and fertilizer rates on forage and grain yields were evaluated in 1989 harvest year for dual production of forage and grain of naked and malting barleys in Cheju province. In the experiment I, 'Saessalbori' (naked barleY)1 and 'Doosan 22' (malting barley) were seeded on Sept. 21, Oct. 12 and Nov. 2, respectively. An unclipped(grain-only) treatment that was planted on Nov. 2 was included for check. In the experiment II, the two above cultivars were planted at seed rate of 14, 20 and 26 and kg/l0a. Fertilizer rates were 9-6-12, 17-13-8 and 22-16-11 (N-P$_2$O$\sub$5/-K$_2$O) kg/l0a. Seeding dates were Oct. 13 and Nov. 7 for clipped treatments and grain -only treatment(seeding rate: 14kg/l0a, N-P$_2$O$\sub$5/-K$_2$O=9-6-12 kg/l0a), respectively. All treatments in both experiments were harvested for grain yield. In Saessalbori, survival following forage removal was 100% regardless of seeding and clipping termination dates, and seeding and fertilizer rates. In Doosan 22, survival percenage decreased with increasing seeding and fertilizer rates when plants was clipped in January to Febuary. Forage yield of two cultivars increased with early planting and delays in clipping termination and tended to increase as seeding and fertilizer rates were increased. Crude ash and fat were not affected by seeding and fertilizer rates, and clipping date. Crude protein increased with an increase in fertilizer rate and decreased with delays in clipping date. N free extract tended to increase with delaying clipping date and decreased with increasing fertilizer rate. Earlier planting resulted in earlier heading and maturity. Clipping slightly delayed heading of Saessalbori and greatly delayed that of Doosan 22. Maturity of Doosan 22 was delayed 3 to 5 days by increased fertilization. Clipping shortened culm length more severely in Doosan 22 than in Saessalbori. The later the clipping termination, the shorter the culm length. The number of spikes per m$^2$ and the number of kernels per spike were not affected by clipping in Saessalbori while those of Doosan 22 were decreased with delays in clipping termination. Delaying forage harvest resulted In a reduction in grain yield. However, final clipping on Feb. 27 reduced grain yield of Saessalbori by only 6-11 % compared to the grain-only treatment. In Doosan 22, forage harvest after Dec. 28 resulted in 6 to 66% reduction of grain yield. The data indicate that grazing of naked and malting barleys until late Feb. and late Dec., respectively, might not reduce grain yields when planted on mid-Sept. to mid-Oct.
Nine plant foods (persimmon leaf, perilla seed, Chinese quince, ginger root, walnut, mugwort leaf, arrowroot, buckwheat and sorghum) rich in phenolic substances were examined for their effects on the digestive enzymes, food-poisoning bacteria and mutagenicity/antimutagenicity by Ames test. Among tested samples, Chinese quince significantly inhibited the $\alpha-amylase$ activity (97%), exhibiting an uncompetitive inhibition type. Protease activity was inhibited by Chinese quince (86%), persimmon leaf (51%) and mugwort leaf (20%), in which mugwort extract exhibited a noncompetitive type. Lipase was activated >50% by all samples. The inhibition of $\alpha-amylase$ was highly correlated with the content of condensed tannin (r=0.89) and the inhibition of protease, with total phenolic content (r=0.84). Total phenolies fraction of tested samples showed the growth inhibition toward E. coli. Streptococcus faecalis and Salmonella enteritidis, in which the effect of perilla, sorghum and arrowroot was the highest for E. coli. Standard phenolics and food samples did not show any mutagenicity toward Salmonella typhimurium TA98 and TA100. Tannic acid inhibited the mutation of the two strains by benzo[a]pyrene whereas total phenolics fractions of Chinese quince and walnut exhibited antimutagenicity to a lesser extent.
Journal of the Korean Society of Food Science and Nutrition
/
v.38
no.3
/
pp.384-390
/
2009
Extraction characteristics of citron (Citrus junos Sieb. ex Tanaka) seeds and functional properties of corresponding extract were monitored by response surface methodology (RSM). Maximum extraction yield of 20.23% was obtained at extraction temperature of $50.23^{\circ}C$, extraction time of 3.03 hr, and shaking velocity of 400.06 rpm. At extraction temperature, extraction time, and shaking velocity of $49.88^{\circ}C$, 2.72 hr, and 400.39 rpm, respectively, maximum polyphenol content was 4.37 mg/g. At extraction temperature, extraction time, and shaking velocity of $50.28^{\circ}C$, 3.42 hr, and 399.96 rpm, respectively, maximum electron donating ability (EDA) was 49.69%. Maximum nitrite scavenging activity (NSA) was 47.79% at extraction temperature, extraction time, and shaking velocity of $49.19^{\circ}C$, 0.68 hr, and 602.95 rpm, respectively. Based on superimposition of 3-dimensional RSM with respect to extraction yield, polyphenol, EDA, and NSA, optimum ranges of extraction conditions were extraction temperature of $50^{\circ}C$, extraction time of about 3 hr, and shaking velocity of 400 rpm.
Huang, Ying;So, Yun-Ji;Hwang, Jeong Rok;Yoo, Kyung-Mi;Lee, Ki-Won;Lee, Yong-Jik;Hwang, In Kyeong
Korean Journal of Food Science and Technology
/
v.46
no.4
/
pp.395-403
/
2014
This study aimed to investigate the activities of phytochemical and bioactive components from 24 new varieties of red pepper obtained by crossbreeding different Korean red pepper lines. The ASTA (American Spice Trade Association) color values ranged from 116.69 to 249.83 and the total carotenoid content ranged from 3.19 to 6.11 mg/g dried red pepper. The free sugar content ranged from 43.41 to 199.54 mg/g, and the fructose content was found to be higher than the glucose content. The ascorbic acid, total polyphenol, and flavonoid contents were found to be 8.08-11.53 mg/g, 7.93-14.26 mg gallic acid/g, and 4.24-10.07 mg quercetin/g dried pepper, respectively. Overall, the total polyphenol contents of the new varieties were found to be higher than the average content of Korean red peppers. The total capsaicinoid content in the new varieties varied widely from 0.42 to 237.87 mg/100 g dried pepper. The pepper-methanol extract antioxidant activities of 'variety 2, 4, 8, 9' were higher than others. A high correlation was observed between the antioxidant activities and the polyphenol contents (r=0.804, r=0.701).
Cornus officinalis Siebold & Zucc. is traditionally used as an edible and medicinal plant in many countries in East Asia. Previous studies have shown the pharmacological potential of extracts and components of C. officinalis, but comparative analysis of the composition of the leaf, stem, and fruit extracts has been insufficient to date. In the present study, the content of active antioxidant and anti-inflammatory ingredients was verified in different C. officinalis parts (under-ripe sansuyu, ripe sansuyu, seed, leaf, stem, and dried sansuyu). One active component, morroniside, was high in fruit (under-ripe and ripe sansuyu), while loganin was high in fruit (under-ripe sansuyu) and cornin was high in seeds. Total polyphenol contents were highest in fruit (ripe sansuyu) and flavonoids were highest in leaves. DPPH radical scavenging activity was highest in leaves, followed by seeds and then ripe sansuyu extract. The anti-inflammatory efficacy of leaf extracts of C. officinalis (LCO) was further investigated by measuring their effects on levels of nitric oxide (NO) and the pro-inflammatory cytokines interleukin (IL)-1β and IL-6 in RAW 264.7 macrophages. Non-cytotoxic concentrations of LCO effectively decreased the lipopolysaccharide (LPS)-induced expression of inducible NO synthase, resulting in decreased NO production. LCO also significantly suppressed LPS-induced production and expression of IL-1β and IL-6. Taken together, the present findings suggest that C. officinalis leaves have potential as natural materials for the development of antioxidant and anti-inflammatory agents.
Lee, Jong-Du;Hyun, Ho Bong;Hyeon, Hyejin;Jang, Eunbi;Ko, Min-Hee;Yoon, Weon-Jong;Ham, Young Min;Jung, Yong-Hwan;Choi, Hwon;O, Eu Gene;Oh, Daeju
Korean Journal of Plant Resources
/
v.35
no.4
/
pp.435-444
/
2022
Hibiscus hamabo Sieb. et Zucc. (yellow hibiscus) is a deciduous semi-shrub plant and mainly growing in Jeju Island. This is known the unique wild hibiscus genus and classified as an 2nd grade of endangered plant for Korean Red List. In previous studies, properties of germination, ecological, genetical and salt resistance have been reported. In this study, we investigated mass-proliferated adventitious root using bioreactor, antioxidant and whitening effects to conduct functional ingredients. Yellow hibiscus were collected from Gujwa, Jeju by prior permission and they were introduced by explant type and various medium composition after surface sterilization. As a result, seed response rates were evaluated at range of 51.17~51.83%, in terms of comprehensive efficiency of shoot and root formation. In the case of adventitious root propagation condition was confirmed in half strength Murashige and Skoog medium salts, 30 mg/L sucrose, and 2 mg/L indole-3-butyric acid for 8 weeks in 5,000 mL bioreactor. We also compared between relationship with biomass and secondary metabolites accumulation by total phenolics content, the flavonoid content, DPPH free radical scavenging activity and melanin content. The results indicated that adventitious root mass proliferation, antioxidant and whitening effect could develop value of the high-quality cosmeceutical ingredient and further metabolite studies.
1. Isolation and identification of amylase-producing bacteria. The powerful strain A-12 and S-8 were respectively isolated from air and soil after screening a large number of amylase-producing bacteria. Their bacterial characteristics have been investigated and it has been found that all characteristics of strain A-12 and S-8 are similar to Bac. subtilis of Bergey's manual except for the acid formation from a few carbohydrates and the citrate utilization, i.e., the strain A-12 shows negative in the citrate utilization, and the acid formation from arabinose and xylose, S-8 shows negative in the acid formation from xylose. 2. Amylase production by Liquid cultures with solid materials. Several conditions for amylase production by strain A-12 in stationary cultures have been studied. The results obtained are as follows. (1) The optimum conditions are:temperature $35^{\circ}C$, initial pH 6.5 to 7.0 and incubation time 3 to 4 days. (2) The amylase production is not affected by the preservation period of the stock cultures. (3) Among the various solid material, the defatted soy bean is found to be the best for t1e amylase production. However, the alkali treatment of the defatted soy bean gives no effect contrary to the cage of defatted rape seed. The addition of soluble starch to the alkali extract of defatted soy bean shows the increased amylase production. (4) Up to 1% addition of ethanol to carbon dificient media gives the improved amylase production, whereas the above effect is not found in the case of carbon rich media. (5) The amylase production can be increased 2.5 times when 10% of defatted soy bean is admixed to cheaply available wheat bran. (6) The excellent effect is found for amylase production when 20% of wheat bran is admixed to defatted dry milk which is a poor medium. The activity is found to be $D^{40^{\circ}}_{30'}$ 7,000(L.S.V. 1,800) in 10% medium. (7) No significant effect is observed due to the addition of various inorganic salts. 3. Amylase production by solid cultures. Several conditions for amylase production by strain A-12 in wheat bran cultures have been studied and the results obtained are as follows. (1) The optimum conditions: are temperature $33^{\circ}C$, incubation lime 2 days, water content added 150 to 175% and the thickness of the medium 1.5cm, The activity is found to be $D^{40^{\circ}}_{30'}$ 36,000(L.S.V. 15,000) (2) No significant effect is found in the case of the additions of various organic and inorganic substances.
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