• KSBB Journal
• /
• v.21 no.6 s.101
• /
• pp.466-473
• /
• 2006

Impaired insulin secretion from pancreatic beta-cells in response to glucose is an important feature in the pathology of non-insulin-dependent diabetes mellitus (NIDDM). In the course of screening for useful insulin secretagogues, we have isolated and identified YHB-2017 (Genistein) as a insulin secretion potentiator from fermentation broths of our in-house microbial library. The insulinotropic activity of YHB-2017 in isolated rat pancreatic islets was exerted only at high concentration of glucose (8.3-16 mM) but not at low concentration of glucose (3.3-5.5 mM). Also, in perifusion study with isolated rat pancreatic islets, YHB-2017 stimulated insulin secretion in a time-dependent manner when YHB-2017 was added to KRB buffer containing 16 mM glucose. In the presence of $200\;{\mu}M$ diazoxide and 35 mM KCI, which stimulates maximum $Ca^{2+}$ influx independently of KATP channel, YHB-2017 enhanced KATP channel-independent insulin secretion at high concentration glucose (16 mM). To elucidate the mechanisms of the glucose-dependent potentiation effect of YHB-2017, pharmacologic inhibitors for protein kinase A, protein kinase C and calcium/calmodulin kinase II were pre-treated and then the potentiation effect of YHB-2017 on insulin secretion was investigated. Pre-treatment of H89 as a PKA inhibitor had a significant inhibitory effect on YHB-2017-induced potentiation effect. Furthermore, western immunoblotting analyses revealed that YHB-2017 increased phosphorylation of PKA substrates and cAMP response element-binding protein (CREB) under high concentration of glucose. These results demonstrated that the insulinotropic effect of YHB-2017 is mediated through PKA signal pathway and activated amplifying $K_{ATP}$ channel-independent insulin secretion pathway.

• Korean Journal of Microbiology
• /
• v.42 no.2
• /
• pp.160-163
• /
• 2006

Brown rice contains rice bran and germ with higher nutritional value and dietary fiber content compared with the polished rice. However, brown rice has a limitation of poor digestion. fermented brown rice could be better nutritional source and improve digestibility. Therefore, we tried to select good fermentative microorganisms which have nutritional values with high amylase and protease activities, and probiotic effects. Nineteen micro-organisms, including eight Bacillus strains isolated from Chongkukjang and 11 lactic acid bacteria, were screened for the fermentation ability and enzyme production. The liquid broths containing 2.5%(w/v) of raw brown rice powder as a sole nutritional source were used for culture media. Among the strains tested, all of the Bacillus strains and two lactic acid bacteria (Leuconostoc gelidum and Pediococcus pentosaceus) showed increase in cell population and enzyme activities. The viable cell counts of all the Bacillus strains and two lactic acid bacteria exceeded $10^7 CFU/mL$. The maximal enzyme activities produced by Bacillus sp. Bl, Bacillus sp. B2, Bacillus sp. B11, L. gelidum and P. pentosaceus were 17.85, 17.50, 17.10, 17.10 and 3.24 U/mL for amylase and 22.48, 22.04, 23.76, 12.13, and 3.4 U/mL for pretense, respectively. Therefore, the results of this study demonstrated that the above strains could be potential starters for the fermentation of raw brown rice.

• Research in Plant Disease
• /
• v.21 no.2
• /
• pp.50-57
• /
• 2015

Control of nematode has become difficult owing to the restricted use of effective soil fumigant, methyl bromide, and other non-fumigant nematicides. Therefore, it is urgently necessary to develop microbial nematicide to replace chemical nematicides. In this study, the 50% aqueous methanol extraction solution of fermentation broths of 2,700 actinomycete strains were tested for their nematicidal activity against second stage of juveniles (J2s) of Meloidogyne incognita. As the results, only the 50% aqueous methanol extraction solution of AN110065, at 20% equivalent to 10% fermentation broth, showed strong nematicidal activity with 78.9% of mortality 24 h after treatment and 94.1% of mortality at 72 h. The 16S rRNA gene sequencing showed that the strain sequence was 99.78% identical to Streptomyces netropsis. The extract of S. netropsis AN110065 fermentation broth was successively partitioned with ethyl acetate and butanol and then the ethyl acetate, butanol and water layers were investigated for their nematicidal activity against the M. incognita. At $1000{\mu}g/ml$, ethyl acetate layer showed the strongest activity of 83.5% of juvenile mortality 72 h after treatment. The pot experiment using the fermentation broth of AN110065 on tomato plant against M. incognita displayed that it evidently suppressed gall formation at a 10-fold diluent treatment. The tomato plants treated with the fermentation broth of S. netropsis AN110065 did not show any phytotoxicity. The results suggest that S. netropsis AN110065 has a potential to serve as microbial nematicide in organic agriculture.

• Korean Journal of Food Science and Technology
• /
• v.48 no.4
• /
• pp.361-365
• /
• 2016

This study was conducted to isolate and identify a microorganism that increases tricin-O-(threo-${\beta}$-guaiacylglyceryl) ether (TTGE) content in the hulls of rice (Oryza sativa L.). Bacteria from germinated rice were isolated by enrichment cultivation using yeast mold, luria bertani, potato dextrose and mannitol egg york polymyxin broths. The highest increase in TTGE content ($339.30{\mu}g/g$) was achieved by a microorganism isolated by PDA enrichment cultivation. On the basis of 16S RNA sequence homology and phylogenetic analysis, the isolated bacterium was identified to have 100% similarity with Burkholderia vietnamiensis. The isolated bacteria were short rods, negative for the Gram stain, and positive for the catalase test. The highest TTGE level was $435.86{\mu}g/g$ in 72-h fermented samples, representing a 2.5x increase compared with the control ($175.65{\mu}g/g$). In conclusion, the bacterium isolated from germinated rice extract was Burkholderia vietnamiensis, and the optimum fermentation period to maximize TTGE levels was 72 h. These findings might help in developing functional materials using rice hulls, a waste product of rice milling.

• Microbiology and Biotechnology Letters
• /
• v.32 no.1
• /
• pp.78-83
• /
• 2004

In order to screen microorganisms producing phopholipase D (PLD) had high transphosphatidylation activity, about 1,000 Actinomycetes strains were isolated from the 63 soil samples, collected over 6 local area in Korea. When the hydrolytic activity in the supernatant was determined, 131 strains produced PLD more than 0.3U/$m\ell$. Among 131 culture broths tested, 23 ones had transphosphatidylation activity higher than 20% and finally one strain (Actinomycetes KF923), which had highest hydrolytic and transphophadylation activity, was selected. Actinomycetes KF923 showed the highest hydrolytic activity (13U/$m\ell$) and phosphatidylation activity (95%) after 48 h fermentation using the P medium (yeast extract 1%, peptone 1%, glucose 1.5%, glycerol 1%, $CaCO_3$ 0.4%, pH 7.2). PLD was purified from the culture broth of Actinomycetes KF923 and the specific activity of purified PLD was 567U/mg. The molecular weight of PLD was about 55kD and the optimum pH and temperature were pH 6.0 and $60^{\circ}C$, respectively. The stability of PLD toward pH and temperature were high around pH 8.0 and below $40^{\circ}C$ Special metal ions were not necessary to the PLD activity.

• Microbiology and Biotechnology Letters
• /
• v.31 no.4
• /
• pp.389-394
• /
• 2003

In order to screen microorganisms producing phopholipase D (PLD) had high transphosphatidylation activity, about 1,000 Actinomycetes strains were isolated from the 63 soil samples, collected over 6 local area in Korea. When the hydrolytic activity in the supernatant was determined, 131 strains produced PLD more than 0.3 U/ml. Among 131 culture broths tested, 23 ones had transphosphatidylation activity higher than 20% and finally one strain (Actinomycetes KF 923), which had highest hydrolytic and transphophadylation activity, was selected. Actinomycetes KF923 showed the highest hydrolytic activity (13 U/ml) and phosphatidylation activity (95%) after 48 h fermentation using the P medium (yeast extract 1%, peptone 1%, glucose 1.5%, glycerol 1%, $CaCo_3$ 0.4%, pH 7.2). PLD was purified from the culture broth of Actinomycetes KF923 and the specific activity of purified PLD was 567 U/mg. The molecular weight of PLD was about 55 kD and the optimum pH and temperature were 6.0 and $60^{\circ}C$, respectively. The stability of PLD toward pH and temperature were high around pH 8.0 and below $40^{\circ}C$. Special metal ions were not necessary to the PLD activity.

• The Korean Journal of Mycology
• /
• v.32 no.1
• /
• pp.8-15
• /
• 2004

This study was conducted to discover new phytotoxins which may be used as lead molecules for the development of new herbicides. A total of 187 endophytic fungi were isolated from 11 plant species, which were collected from 8 locations in Korea. Their herbicidal activities were screened in vivo by herbicidal and duckweed bioassays after they were cultured in potato dextrose broth and rice solid media. Both fermentation broth and solid culture extract of Gliocladium catenulatum F0006 isolated from red pine (Pinus densiflora) showed 70% herbicidal activity only against cocklebur (Xanthium strumarium) out of the 10 weeds tested. Solid culture extract of F0034 isolated from arrowroot (Pueraria thunbergiana) exhibited 20 to 100% herbicidal activities against all of the weeds. Especially, shattercane (Sorghum bicolor), barnyardgrass (Echinochloa crus-galli), large crabgrass (Digitaria sanguinalis), and fall pauicum (Panicum dichtomiflorum) were sensitive to the solid culture extract of F0034. In addition, solid culture extract of F0043 isolated from red pine displayed 20% to 70% herbicidal activities only against 5 grass species, but not against 5 broad-leaf plant species. On the other hand, as the results of duckweed assay, 8 fermentation broths showed 100% growth inhibitory activity at concentrations less than 5.0% of culture supernatants and 12 solid cultures had a potent inhibitory activity against duckweed growth. A toxic metabolite was purified from the solid cultures of G. catenulatum F0006 by repeated column chromatography and bioassay. It caused a phytotoxic syndrome only on cocklebur out of the 10 weeds tested; it completely killed cocklebur seedlings at $500\;{\mu}g/ml$ and showed 85% herbicidal activity against cocklebur at $100\;{\mu}g/ml$. The molecular weight of the toxic metabolite is 238 daltons and its structure determination is underway.

• Research in Plant Disease
• /
• v.15 no.2
• /
• pp.120-126
• /
• 2009

In the course of study on the roles of phytotoxins in the pathogenicity of Botrytis cinerea, we isolated two novel phytotoxins. They were identified as 3-O-acetyl botcinol and 3-O-acetyl botcinolide. In this study, we investigated correlation between the two phytotoxins and the pathogenicity of B. cinerea. In liquid cultures, the two phytotoxins were not produced by three low pathogenic isolates out of 25 B. cinerea isolates. Among strong or moderate pathogenic isolates, some produced the two phytotoxins, but the others did not. On the other hand, the ethyl acetate extracts of fermentation broths of 10 out of 25 isolates showed phytotoxic activity against various plants tested in a whole plant assay. The phytotoxins were detected in all of the 10 phytotoxic ethyl acetate extracts. In planta, the two phytotoxins were detected in all of the plant tissues infected with strong pathogenic isolates. However, there was no correlation between the ability of B. cinerea isolates to produce the two phytotoxins and their pathogenicities. The two phytotoxins began to detect in tomato plant tissues infected with B. cinerea 2-16 at 3 days after inoculation, increased gradually till 4 days after inoculation, and then decreased. The above results suggest that 3-O-acetyl botcinol and 3-O-acetyl botcinolide are one of pathogenicity factors for B. cinerea, but not a primary determinant of its pathogenicity.