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

• Korean Journal of Microbiology
• /
• v.31 no.3
• /
• pp.218-223
• /
• 1993

Conditions for the release and regeneration of protoplasts form Rhizopus oryzae and intergeneric protoplast fusion between Rhizopus oryzae and Aspergillus oryzae were studied. High yields of protoplast fusion between Rhizopus oryzae and Aspergillus oxyzae were studied. High yield of protoplasts from young germilings of R. oryzae were obtained by using lytic enzymes containing chitosanase (3 mg/ml), chitinase (3 mg/ml) and Novozym 234 (5 mg/ml). 0.5M glucose was used as the osmotic stabilizer and optimum pH of buffer was determined to be pH 7.5-8.0. Under these conditions, protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts regenerated on solid medium with a soft agar overlay. We have also carried out protoplasts fusion between R. oryzae and A. oryzae and have succeeded in obtaining three types of intergeneric fusants. In these experiments, 35% PEG-4000 and 10 mM CaCl$_{2}$ were used as fsogenic agents, and auxotrophic properties were used as a genetic marker to select fusants. Complementation frequency be protoplasts fusion of A. oxyzae and R. oryzae was 4.4% * 10$^{-5}$ . The fusant strains of the first type were prototrophs showing an Aspergillus type morphology with dark-yellow sporulation, those of the second type were also Apergillus type morphology but showed no sporulation. And the strains of the third type stopped growing when fusion products grown on regeneration minimal medium were transferred to fresh minimal medium. The formation of fusion products was observed by fluorescent vital stains for complementary labelling of protoplats from R. oryzae and A. oryzae. Rhodamine 6G and fluorescein diacetate wer useful complementary vital stains of Rhizopus and Aspergillus protoplasts for visualization of requency and type (dicell, multicell) of fusion.

• Journal of Microbiology and Biotechnology
• /
• v.11 no.1
• /
• pp.7-12
• /
• 2001

Three Pseudomonas strains (Bk8, Bk9, Bk10) selected from soil for their ability to degrade herbicide diuron were tested for their heavy metal resistance. The growth of these catabolic strains on a minimal medium with various concentrations of $Cd^{2+},\;Zn^{2+},\;Ni^{2+}$, and $Hg^{2+}$ revealed a minimal effect on the carbon source for the inhibitory effect of the metals. One of these strains, namely, Bk8, exhibited a high resistance to the heavy metals as compared to the two other strains. This strain harbors plasmid pBk8 (110 kb) and contains at least fur determinants encoding heavy metal resistance. Nickel and zinc resistance are encoded by genes located on the chromosome, while cadmium and mercury resistance are on plasmid pBk8. Accordingly, the characteristics of strain Bk8 suggest that it would be useful in the bioremediation of aromatic compounds in the presence of toxic heavy metals as co-contaminants.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.6
• /
• pp.849-854
• /
• 2006

The effects of Haematococcus pluvialis extracellular products on microbial growth and bacteriocin production were investigated to improve bacteriocin synthesis during the growth cycle of Lactobacilli. Lactobacillus pentosus KJ-108, L. plantarum KJ-10311, and L. sakei KJ-2008 were cultured in MRS and enriched medium (ERM) with or without supplement of the extracellular products obtained from a late exponential phase culture of Haematococcus pluvialis in modified Bold's basal medium (MBBM). In both MRS and ERM, the extracellular products strongly enhanced the growth as well as the bacteriocin production of all the lactic acid bacteria tested. The enhancing effect was observed in ERM with pH adjusted at 5 and 6. In addition, some difference in growth effects with the extracellular products of H. pluvialis was observed between pH 5 and 6 in ERM, but no effect was observed in the minimal medium. The final biomass and the final concentration of bacteriocin activity were associated with the cell growth that was promoted by the extracellular products of H. pluvialis, and the enhanced cell growth of the three lactic acid bacterial strains induced the increase of the specific bacteriocin production. Therefore, bacteriocin production and activity were influenced by the addition of the extracellular products of H. pluvialis in the culture medium.

• Proceedings of the Korean Biophysical Society Conference
• /
• 2003.06a
• /
• pp.61-61
• /
• 2003

We have cloned the CGR1 gene encoding glutathione reductase (GR) which catalyzes the reduction of oxidized glutathione (GSSG) to reduced glutathione (GSH) from Candida albicans. The cgr1/cgr1 mutants were not viable when CaMAL2 promoter repressed the CGR1 expression. The growth of the mutants could be partially overcome by thiol compounds such as GSH, dithiothreitol, cysteine, N-acetylcysteine and GSSG. Interestingly, C. albicans with CGR1 overexpressed showed defective hyphal growth on solid medium and attenuated virulence. We have also cloned the GCS1 gene encoding ${\gamma}$-glutamylcysteine synthetase which catalyzes the first step of glutathione biosynthesis. The gcs1/gcs1 mutants were nonviable in minimal defined medium. The growth of the mutants could be resumed by supplementing with GSH, GSSG and ${\gamma}$-glutamylcysteine in the medium. The mutants had increased intracellular D-erythroascorbic acid level up to 2.25-fold when transferred to GSH-free medium. When the mutants were depleted of GSH, they showed typical markers of apoptosis. In conclusion, these results suggest that glutathione is an essential metabolite, and involved in hyphal growth, virulence and apoptosis in C. albicans.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.7 no.3
• /
• pp.163-170
• /
• 2002

This report describes the use of a transtubular bioreactor to study the relative effects of diffusion versus perfusion of medium on antibody production by a hybridoma cell line. The study was performed with a high-density cell culture maintained in a serum-free, low-protein medium for 77 days. It was determined that the reactor possessed a macro-mixing pattern residence time distribution similar to a continuous stirred tank reactor (CSTR), However, due to the arrangement of the medium lines in the reactor, the flow patterns for nutrient distribution consist of largely independent medium path lengths ranging from short to long. When operated with cyclic, reversing, transtubular medium flow, some regions of the reactor (with short residence times) are more accessible to medium than others (with long residence times). From this standpoint, the reactor can be divided into three regions: a captive volume, which consists of medium primarily delivered via diffusion; a lapped volume, which provides nutrients through unilateral convection; and a swept volume, which operates through bilateral convection. The relative sizes of these three volumes were modified experimentally by changing the period over which the direction of medium flow was reversed from 15 min (larger captive volume) to 9 h (larger swept volume). The results suggest that antibody concentration increases as the size of the diffusion-limited (captive) volume is increased to a maximum at around 30 min with a sharp decrease thereafter. As reflected by changes in measured consumption of glucose and production of lactate, no significant difference in cellular metabolism occurred as the reactor was moved between these different states. These results indicate that the mode of operation of the transtubular bioreactor may influence antibody productivity under serum-free, low-protein conditions with minimal effects on cellular metabolism.

• Korean journal of applied entomology
• /
• v.50 no.4
• /
• pp.307-314
• /
• 2011

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 $25^{\circ}C$ 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.

• The Korean Journal of Food And Nutrition
• /
• v.1 no.1
• /
• pp.25-32
• /
• 1988

Properties of purified isocitrate lyase from Saccharomycopsis lipolytica ATCC 44601 and MX9-11RX8 temperature-sensitive mutant were investigated. Purified isocitrate lyase from temperature-sensitive mutant was indistinguishable from the wild type enzyme with respect to the isoelectric pH(5.3), the thermostability and Km value for threo-Ds-Isocitrate(about 0.2 mM). When isocitrate lyase induced by acetate minimal medium at 33$^{\circ}C$, MX9-11RX8 mutant did not express enzyme activity but did synthesize polypeptide chain whose electrophoretic mobilities were equal to those of the purified enzymes.

• Journal of Microbiology and Biotechnology
• /
• v.5 no.1
• /
• pp.48-50
• /
• 1995

We discovered that the Bacillus licheniformis SSA3, fermenting traditional Korean soy sauce and soybean paste, involved in the synthesis of a dark-brown pigment. This pigment produced in the minimal medium supplemented with tyrosine only as precursor. We showed that this pigment is novel, and differed from melanoidin and melanin, and an antimutagenic substance.

• Mycobiology
• /
• v.33 no.1
• /
• pp.12-14
• /
• 2005

The culture conditions for the enhanced mycelial growth of Ramaria botrytis was investigated. The optimal temperature and pH for the mycelial growth were $24^{\circ}C$ and 5.0, respectively. It was shown that starch was best of several carbon sources in Czapek-Dox medium as a minimal medium for the enhanced mycelial growth. Organic nitrogen sources were better than inorganic ones for mycelial growth. The appropriate vitamin and mineral salt were biotin and FeCl3, respectively. When this strain was cultured with $FeCl_3$ for 30 days, 19.23 g/l of dry mycelium of R. botrytis was obtained.