• Korean Journal of Medicinal Crop Science
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• v.15 no.5
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• pp.346-351
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• 2007

An efficient somatic embryogenesis and plant regeneration protocol was developed for Schisandra chinensis Baill, using embryogenic cell suspensions and optimized media conditions. Friable embryogenic callus was induced from cotyledonary leaf and hypocotyl explants of 7 days old seedlings on MS agar medium supplemented with 1.0 to $4.0\;mg\;l^{-1}$ of 2,4-dichlorophenoxyacetic acid (2,4-D). Fast growing and well dispersed embryogenic cell suspensions were developed within two months when embryogenic calli were transferred to MS liquid medium containing $1.0\;mg\;l^{-1}\;2,4-D$. One third strength of MS medium was the best for both overall growth and development of somatic embryos in liquid culture. Over 3400 viable somatic embryos were produced from each 150 ml flask with an initial cell density of 30 mg in 30 ml medium. Germinated somatic embryos developed in liquid medium converted into plantlets after transferred to half-strength MS semi-solid medium. Approximately 90% of the converted plantlets were successfully transplanted to soil and grew into fertile plants.

• Korean Journal of Medicinal Crop Science
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• v.20 no.6
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• pp.479-486
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• 2012

The production of adventitious roots derived from root explant of Echinacea angustifolia and its secondary metabolite content were assessed in different types and levels of auxin. The induction of adventitious roots from root explant cultured in Murashige and Skoog solid medium supplemented with 1.0 mg/L indole -3-butyric acid (IBA) attained highest as 20.87 mg fresh weight and 3.07 mg dry weight per culture but root suspension culture at the same concentration of IBA enhanced biomass production as 3.07 g fresh weight and 0.38 g per culture after 4 weeks in culture. 3.0 mg/L ${\alpha}$-naphthalene acetic acid (NAA) treatment had similar effect on root biomass production as 3.07 g fresh weight and 0.38 g per culture with liquid suspension culture, whereas adventitious roots exposed to over 3.0-5.0 mg/L IBA or 5.0 mg/L NAA were less responsive by reducing the number of adventitious roots and/or changing root morphology such as short and thick. The content of secondary metabolites such as phenolic, flavonoids and total caffeic acid in adventitious roots cultured on MS medium supplemented with 1.0 mg/L IBA were attained highest as 27.20, 9.60. 10.67 mg/g dry weight, respectively. Overall, the best production of root biomass and secondary metabolites were given by 1.0 mg/L IBA.

• Proceedings of the Korean Institute of Resources Recycling Conference
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• 2001.05b
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• pp.121-125
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• 2001

For the probiotic feed production, aerobic liquid fermentation of pulverized food wastes was attempted with a yeast Kluyveromyces marxianus. After grinding finely, optimal fermentation conditions of the substrate was investigated by shaking culture. The most active growth of the yeast was shown at solid content of 10%. The proper addition of urea(0.5g/l), o-phosphate(0.4g/l), molasses(4g/l), and yeast extract (1g/1) increased cell growth rate and viable cell count. For optimizing, the nutrients were all added to substrate and fermentation was carried in 2 litre jar fermenter. For the stimulation of hydrolyzing enzyme excretion, mixed culture with Aspersillus oryzae was also conducted. In 12 hours of fermentation, viable cell count of the yeast Kluyveromyces marxianus amounted to the number of 1.4 $\times$10$^{10}$ /1 in the culture medium.

• Tuberculosis and Respiratory Diseases
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• v.53 no.6
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• pp.635-643
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• 2002

Background : The Aim of this study was to compare the recovery of mycobacteria from sputum samples of pulmonary tuberculosis patients using the MB/BacT rapid culture system(Organon Teknika, USA) with that obtained using Lowenstein-Jensen solid medium. Methods : The two culture systems were compared using sputum samples of 99 pulmonary tuberculosis patients. Culture media were incubated at $35-37^{\circ}C$ for six weeks in the MB/BacT system and for 12 weeks in Lowenstein-Jensen solid medium. Solid media were examined macroscopically once a week, and the MB/BacT system positive vials were unloaded from the machine as soon as possible after positive signal from the connected computer was detected Confirmation of growth for mycobacteria was done by Ziehl-Neelson stained smears. Isolates were identified to differentiate Mycobacterium tuberculosis from mycobacterium other than tuberculosis(MOTT) by phenotypic and molecular methods. Results : Of the sputum samples of the 99 patients, 58 samples were smear positive and 41 in negative smear. Mycobacteria were recovered from 67(67.7%) samples by using both culture systems. The yield with MB/BacT was higher than that with Lowenstein-Jensen [67(67.7%) vs. 52(52.5%), p<0.001]. Moreover, 15(15.2%) samples were positive only in the MB/BacT, whereas none of samples was positive only in Lowenstein-Jensen. In smear-positive and smear-negative samples, the recovery rate with MB/BacT was also higher than that with Lowenstein-Jensen [sputum-positive; 56/58(96.6%) vs. 46/58(79.3%), p=0.005, sputum-negative; 6/41(14.6%) vs. 5/41(12.2%), p<0.001]. The mean times to detection of Mycobacteria were 13.3 and 27.2 days with MB/BacT and Lowenstein-Jensen respectively(p<0.001). Conclusion : This results indicate that the the MB/BacT is more efficient and faster than Lowenstein-Jensen for the recovery of mycobacteria.

• Mycobiology
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• v.28 no.1
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• pp.33-38
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• 2000

Mycelium of Hericium erinaceum isolate KU-1 was cultured in liquid medium (HL medium) and solid medium (Ko medium) at pH 4.0 in $28^{\circ}C$. 1.0% glucose or fructose was the most favorable carbon source, and 0.2% amonium acetate or $NaNO_3$ was an exellent nitrogen source for mycelial growth as well as production of antimicrobial substances. The mixture of saw dust 70% with rice bran 30% (SR medium) was the substrate for formation of sporophores. The active substrates in extracts from mycelium, culture filtrate and fruiting body were separated by TLC. The solvent for TLC was EtOAc: Chloroform: MeOH (10 : 5 : 10). Phenol-like substances appeared at Rf $0.5{\sim}0.9$, and fatty acid-like substances appeared at Rf $0.1{\sim}0.2$. The purified materials from the extracts showed antimicrobial effects to Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Aspergillus niger, Candida albicans and Microsporum gypseum. The S. aureus was the most inhibited. Minimal inhibitory concentration (MIC) of purified white powder and the Hercenone derivatives against S. aureus were $5.65\;{\mu}g/ml$ and $1.85\;{\mu}g/ml$, respectively.

• Journal of Microbiology
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• v.44 no.3
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• pp.276-283
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• 2006

The thermophilic fungus Thermoascus aurantiacus 179-5 produced large quantities of a glucosidase which preferentially hydrolyzed maltose over starch. Enzyme production was high in submerged fermentation, with a maximal activity of 30 U/ml after 336 h of fermentation. In solid-state fermentation, the activity of the enzyme was 22 U/ml at 144 h in medium containing wheat bran and 5.8 U/ml at 48 h when cassava pulp was used as the culture medium. The enzyme was specific for maltose, very slowly hydrolyzed starch, dextrins (2-7G) and the synthetic substrate (${\alpha}$-PNPG), and did not hydrolyze sucrose. These properties suggest that the enzyme is a type II ${\alpha}$-glucosidase. The optimum temperature of the enzyme was $70^{\circ}C$. In addition, the enzyme was highly thermostable (100% stability for 10 h at $60^{\circ}C$ and a half-life of 15 min at $80^{\circ}C$), and stable within a wide pH range.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.1
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• pp.78-84
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• 2005

The purpose of this study was to optimize the solid state cultivation of Monascus ruber on sterile rice. A single-level-multiple-factor and a single-factor-multiple-level experimental design were employed to determine the optimal medium constituents and to optimize carbon and nitrogen source concentrations for lovastatin production. Simultaneous quantitative analyses of the ${\beta}$-hydroxyacid form and ${\beta}$-hydroxylactone for of lovastatin were performed by the high performance liquid chromatography (HPLC) method with a UV photodiode-array (PDA) detector. The total lovastatin yield ($4{\sim}6\;mg/g$, average of five repeats) was achieved by adding soybean powder, glycerol, sodium nitrate, and acetic acid at optimized levels after 14 days of fermentation. The maximal yield of lovastatin under the optimal composition of the medium increased by almost 2 times the yield observed prior to optimization. The experimental results also indicated that the ${\beta}$-hydroxylactone form of lovastatin (LFL) and the ${\beta}$-hydroxyacid form of lovastatin (AFL) simultaneously existed in solid state cultures of Monascus ruber. while the latter was the dominant form in the middle-late stage of continued fermentation. These results indicate that optimized culture conditions can be used for industrial production of lovastatin to obtain high yields.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.57 no.4
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• pp.317-323
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• 2012

For the establishment of an efficient embryogenesis from microspore culture in Brassica napus L. domestic cultivar 'Tammiyuchae', four different factors affecting microspore embryogenesis and plantlet regeneration were investigated. The highest embryogenesis rate was achieved when microspores at late uninucleate to early binucleate stage were isolated from flower buds with a length of 3.0~3.5 mm. On average, 388 embryos generated from 1 ml of microspores media. The highest number of embryos was obtained when microspores were subjected to $32.5^{\circ}C$ for 2 days. Embryogenesis of 'Tammiyuchae' was increased with increasing microspore culture density up to about $5{\times}10^4ea/mL$. Gradually higher culture density repressed embryogenesis of microspores. Regeneration rate of shoots from microspore-derived embryos was observed in MS solid medium supplemented with $0.5mg{\cdot}L^{-1}$ NAA and $1.0mg{\cdot}L^{-1}$ BA, and grew well in MS solid medium without plant growth regulators.

• The Korean Journal of Food And Nutrition
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• v.29 no.1
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• pp.1-11
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• 2016

To enhance the physiological activities of roasted coffee (RC), 30 kinds of green coffee beans (GCB) with different cultivating areas and varieties were fermented with Monascus ruber mycelium (MR) by solid-state culture. After the dried MR-fermented GCB was subjected medium roasting, each RC was extracted with hot-water. Among the hot-water extracts, the highest yield was the hot-water extract of RC from MR-fermented Indonesia Mandheling GCB (15.5%). However, the hot-water extract of RC from MR-fermented Ethiopia Sidamo GCB showed significantly higher polyphenolic contents (3.08 mg GAE/100 mg) and ABTS free radical scavenging activity (25.41 mg AEAC/100 mg). Meanwhile, the hot-water extract of RC from MR-fermented Vietnam Robusta GCB showed not only the effective inhibition of $TNF-{\alpha}$ level (73.7% inhibition of LPS-stimulated control) from LPS-stimulated RAW 264.7 cells but also significant inhibition of lipogenesis (63.5% inhibition of lipid differentiation control) in 3T3-L1 pre-adipose cells. In conclusion, these results suggest that roasted coffees from Ethiopia Sidamo and Vietnam Robusta green coffee beans fermented with Monascus ruber mycelium using solid-state culture could have industrial applications as functional coffee beverages.

• KSBB Journal
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• v.22 no.5
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• pp.305-312
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• 2007

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.