• Food Science and Preservation
• /
• v.12 no.4
• /
• pp.379-386
• /
• 2005

Free amino acids and polysaccharide content of submerged mycelial culture of Fomitopsis pinicola using $2\%$ citrus peel water extracts(CP), $2\%$ green tea water extracts(GT) and yeast-malt broth(YM) were investigated. Cultivations were conducted at $30^{\circ}C$ and 150 rpm for 10 days. Yields of the mycelium as fresh weight basis in GT-, CP- and YM-broth were $42.3\%,\;34.2\%\;and\;9.89\%$, and their turbidity(OD at 660 nm) of the broth without mycelium were 0.14, 0.16 and 0.22, respectively. Total free amino acid content in the mycelium were 928.19 $mg\%$ in YM, 1060.53 $mg\%$ in CP, 764.83 $mg\%$ in GT, and the major free amino acid was lysine in YM, glutamic acid in CP and GT. Total free exo-amino acid contents were 659.75 $mg\%$ in YM, 954.55 $mg\%$ in CP, 838.69 $mg\%$ in GT, and the major in the all broths was glutamic acid Total amino acid derivatives content of the mycelium were in order of CP>GT>YM, and the major was cystathionine in YM, hydroxy proline in CP and GT. The major among exo-amino derivatives was hydroxy proline in the all broth. Exo-AIS content was in order of CP>GT>YM. Acid soluble polysaccharide content of the mycelium was GT($0.69\%$)>YM($0.39\%$)>CP($0.18\%$). The exo-polysaccharide content was in order of GT($0.87\%$)>CP($0.69\%$)>YM($0.09\%$). Alkali soluble polysaccharide content of the mycelium was in order of CP($5.21\%$)>GT($5.18\%$)>YM($4.56\%$), and exo-polysaccharide was in order of GT($6.79\%$)>YM($3.57\%$)>CP($3.01\%$). The alkali soluble polysaccharide eluted from mycelium cultivated in CP broth was supposed to polysaccharide(about 500,000 daltons) composed of hexose and uronic acid bounded with protein(below 10,000 daltons).

• The Korean Journal of Mycology
• /
• v.41 no.4
• /
• pp.236-242
• /
• 2013

It is known not only that antifungal compounds such as sparassol, methyl orsellinate (ScI) and methyl-dihydroxy-methoxy-methylbenzoate (ScII) were produced during submerged culture from Sparassis crispa, but also that ScI and ScII were appeared higher antifungal activity than sparassol. The aim of this study, antifungal compounds of Sparassis latifolia were purified from mycelial culture media and identified by using NMR and ESI-MS. Based on HPLC analysis, methyl orsellinate and sparassol were detected at 15 min and 31 min of retention time, respectively. The compounds derived from S. latifolia were classified into four production patterns according to their strains. The strains originated from host plant Larix kaempferi and Pinus koraiensis showed different patterns of compound production, whereas the strains originated from host plant P. densiflora and Abies holophylla showed almost same patterns. There was no correlation between mycelial biomass and compound production. KFRI 645 strain from L. kaempferi exhibited higher methyl orsellinate production (0.170 mg/ml). Sparassol was produced by KFRI 747 from P. densiflora (0.004 mg/ml). Thus, our result revealed the new fact that methyl orsellinate and sparassol have different patterns according to the strains originated from different host plants.

• KSBB Journal
• /
• v.22 no.1
• /
• pp.30-36
• /
• 2007

The water-soluble or ethanol-soluble materials extracted from fruit bodies and cultured products (mycelium and broth) of H. erinaceum were prepared, and their inhibitory effect on acetylcholinesterase (AChE) activity from Electrophorous electricus was investigated. Inhibition of 75-85% for AChE activity at concentration of 10 mg/ml was obtained and the mechanism was due to general non-competitive inhibition. Especially, the supernatant of culture broth by ethanol treatment, exhibited a strong inhibition activity of 94% at 10 mg/ml. The samples from fruit body, mycelium and broth (supernatant and precipitate by ethanol treatment) which were extracted from H. erinaceum, were very effective to inhibit the initial stage oxidation of a linoleic acid at concentration of 0.1 mg/ml. The antioxidative activity of these samples were superior than rutin, vitamin C and tocopherol as antioxidative standards by FTC (ferric thiocyanate) method, and also showed the very strong antioxidative activity of 95% without significant difference of the samples by TBA-RS (thiobarbituric acid-reactive substance) method.

• Journal of Microbiology and Biotechnology
• /
• v.19 no.9
• /
• pp.987-996
• /
• 2009

Aspergillus niger GH1 previously isolated and identified by our group as a wild tannase producer was grown under solid-state (SSC) and submerged culture (SmC) conditions to select the enzyme production system. For tannase purification, extracellular tannase was produced under SSC using polyurethane foam as the inert support. Tannase was purified to apparent homogeneity by ultrafiltration, anion-exchange chromatography, and gel filtration that led to a purified enzyme with a specific activity of 238.14 IU/mg protein with a final yield of 0.3% and a purification fold of 46. Three bands were found on the SDS-PAG with molecular masses of 50, 75, and 100 kDa. PI of 3.5 and 7.1% N-glycosylation were noted. Temperature and pH optima were 600e and 6.0 [methyl 3,4,5-trihydroxybenzoate (MTB) as substrate], respectively. Tannase was found with a $K_M$ value of $0.41{\times}10^{-4}M$ and the value of $V_{max}$ was $11.03{\mu}$moL/min at $60^{\circ}C$ for MTB. Effects of several metal salts, solvents, surfactants, and typical enzyme inhibitors on tannase activity were evaluated to establish the novelty of the enzyme. Finally, the tannase from A. niger GH1 was significantly inhibited by PMSF (phenylmethylsulfonyl fluoride), and therefore, it is possible to consider the presence of a serine or cysteine residue in the catalytic site.

• Journal of Korean Society of Forest Science
• /
• v.39 no.1
• /
• pp.57-63
• /
• 1978

Enzymatic hydrolysis of the substrate from Alnus hirsuta (Spach) Rupr (8-14years) was investigated using cellulase preparations of Trichoderma viride Pers. ex. Fr. SANK 16374 and conduced on the optimum reaction conditions of the cellulase on saccharification. The crude cellulase was produced by the submerged culture process and produced in the culture fluid was salted out quantitatively by the use of ammonium sulfate. The method of delignification from wood(Saw dust) was treated by the peracetic acid (PA) method. Reducing sugar was determined by the dinitrosalicylic acid (DNS) method. The results were summerized as follows; 1. The optimum pH of cellulase was 5.0 and the range of stability with respect to pH was generally from 4.0 to 6.0 2. The optimum temperature of cellulase was generally $40^{\circ}C$, but reducing sugar formation did not show significent differences at 5% levels in the reaction temperature from $40^{\circ}C$ to $50^{\circ}C$. 3. The redusing sugar were increased with increase of cellulase concentration. 4. The reducing sugar were decreased with increase of substrate concentration. 5. Fructose was a very good inhibitor of the enzyme from Trichoderma viride, but glucose inhibition was generally weak.

• Journal of Life Science
• /
• v.15 no.2 s.69
• /
• pp.287-292
• /
• 2005

The crude galactomannans (GMs) were obtained from the culture of a newly isolated fungus Trichoderma erinaceum DG-312 and their anti-inflammatory activity was investigated in mice. The maximum concentrations of mycelial biomass and GMs reached 9.44 g/l, 2.72 g/l at day 3 in a 5-l stirred-tank bioreactor, respectively. The results of Sepharose CL-6B gel chromatography and compositional analysis revealed that the crude GMs contain heterogeneous polysaccharides consisting of $74.9\%$ mannose and $24.1\%$ galactose. The GMs was shown to possess a significant anti-inflammatory activity against acetic acid-induced inflammatory mouse model in a dose-dependent manner, when mice were treated with 100 and 200 mg GMs/kg body weight. The inhibition in vascular permeability $(60.6\%)$ and in writhing response $(62.5\%)$ evidenced an anti-inflammatory activity of the GMs. The marked anti-inflammatory and writhing-lowering properties of the GMs suggest its potential therapeutic use.

• The Korean Journal of Mycology
• /
• v.15 no.3
• /
• pp.149-157
• /
• 1987

This study was undertaken to investigate the differentiation, from spore germination to hyphae growth and phialide formation, of Aspergillus niger through the method of synchronous and submerged culture. Through continuous experiments by shake culture with potassium acetate medium, we observed the formation of spores at appropriate concentration and pH. Potassium acetate medium was set pH 5.5, 6.0, 6.5 and 7.0 on each scale, and control, 20 mM, and 40 mM, 80 mM and 160 mM concentrations on the other scale. Aspergillus niger was cultured in the defined media at $28^{\circ}C$, and mycelial dry weight, changes of pH and the onset of sporulation were checked. The mycelial dry weight, increased in potassium acetate medium, and pH increased during mycelial growth and gradually decreased after the spore formation. When pH increased excessively in Potassium acetate medium with pH 7.0, the mycelia could not adapt and mycelial dry weight decreased gradually. At pH 5.5, the onset of sporulation was done within one day at 20 mM it took, at 80 mM three days and at 160 mM concentration. in two days, at 40 mM one to four days were taken, 80 mM concentration respectively. At pH 6.5, the onset of sporulation was done in three days and four days at 80 mM concentrations respectively. Spore formation was not shown at pH 7.0. In controlled medium with all levels of pH, spore formation was not shown.

• The Korean Journal of Mycology
• /
• v.15 no.4
• /
• pp.238-246
• /
• 1987

Aspergillus niger van Tieghem was cultured by the method of synchronous and submerged culture. The sporulation occurred through the culture and its life cycle and differentiation were completed in the experiments. The effects of cAMP, theophylline and caffeine on the sporulation of A. niger were investigated. In the sporulation medium, the sporulation was stimulated by addition of cAMP and its optimum concentration was $10^{-4}M$. In the sporulation medium, the sporulation was stimulated by addition of theophylline and its optimum concentration was 10 mg/ml. In the sporulation medium, the sporulation was stimulated by addition of caffeine and its optimum concentration was 300 mg/ml. Theophylline added to the sporulation medium together with cAMP enhanced the promotion effect of cAMP on sporulation. Caffeine added to the sporulation medium together with cAMP enhanced the promotion effect of cAMP on sporulation. In the sporulation medium, the sporulation was stimulated by addition of neither AMP nor ATP. In the potassium acetate medium, cAMP, theophylline and caffeine stimulated the sporulation, respectively.

• Journal of Life Science
• /
• v.16 no.3 s.76
• /
• pp.477-484
• /
• 2006

This study was performed to know the potentialities of the fruits of Opuntia ficus-indica, as a medium for mushroom mycelial culture. Five mushroom mycelial (Agrocus blazei, Grifola frondosa, Hericium erinaceum, Innonotus obliquus, Phellinus linteus) were frown on the malt extract broth (MEB) and the cactus broth medium (CB). The submerged culture mixtures were extracted using equal volume of ethyl acetate, and their extract yields, total polyphenol contents, and some physiological activities were compared with each other Each extract from mycelial culture grown on CB medium showed remarkable enhancement in physiological activities compared with each counterpart grown on MEB. Among five mycelial cultures grown on CB medium, the extract yield and polyphenyl content were highest in the extract from Grifola frondosa (extract yield, 0.4 g/L and polyphenol content, 22.7%). Also, the extracts from Grifola frondosa showed the highest physiological activities, such as DPPH radical scavenging ($IC_{50}=362.9{\mu}g/ml$), xanthine oxidase inhibition (about 80% at $500{\mu}g/ml$), and superoxide radical scavenging (about 80% at $500{\mu}g/ml$), and NO production inhibition ($IC_{50}=43.1{\mu}g/ml$) in LPS-stimulated RAW 264.7 cells. This result suggests that the fruit of Opuntia ficus-indica can be used as a culture medium for improving the functional properties of various mushroom mycelia.

• KSBB Journal
• /
• v.22 no.5
• /
• pp.288-296
• /
• 2007

Monascus pilosus (KCCM 60160) in submerged culture was optimized based on culture medium and fermentation conditions. Monacolin-K (Iovastatin), one of the cholesterol lowing-agent which was produced by Monascus pilosus may maintain a healthy lipid level by inhibiting the biosynthesis of cholesterol. Plackett-Burman design and response surface method were employed to study the culture medium for the desirable monacolin-K production. As a result of experimental designs, optimized production medium components and concentrations (g/L) were determined on soluble starch 96, malt extract 44.5, beef extract 30.23, yeast extract 15, $(NH_4)_2SO_4$ 4.03, $Na_2HPO_4{\cdot}12H_2O$ 0.5, L-Histidine 3.0, $KHSO_4$ 1.0, respectively. Monacolin-K production was improved about 3 times in comparison with shake flask fermentation of the basic production medium. The effect of agitation speed (300, 350, 400 and 450 rpm) on the monacolin-K production were also observed in a batch fermenter. Maximum monacolin-K production with the basic production medium was 68 mg/L when agitation speed was 500 rpm. And it was found that all spherical pellets (average diameter of $1.0{\sim}1.5mm$) were dominant during fermentation. Based on the results, the maximum production of 185 mg/L of monacolin-K with the optimized production medium was obtained at pH (controlled) 6.5, agitation rate 400 rpm, aeration rate 1 vvm, and inoculum size 3%.