• Journal of Plant Biotechnology
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• v.42 no.2
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• pp.104-110
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• 2015

We analyzed the effects of various LED light treatments (red 655 nm, blue 456 nm, white and mixed light) on growth pattern, gene expression and flavonoid contents in lettuce leaf. Plants treated with mixed light (red+blue+white) showed better growth performance than those treated with single LED and fluorescent lamp (FL). Expression analysis of the eight genes involved in flavonoid biosynthesis in plants treated with LED light was examined. Results showed that red lettuce grown under mixed light showed high expression levels of LsC4H, LsF3H and LsDRF genes. Morever, the same treatment plants possessed higher content of gallic acid, chlorogenic acid and quercetin contents than those in plants exposed to single light. However, the highest total anthocyanin content was identified in plants treated with red+blue light and the lowest content was identified in plants exposed to white fluorescent lamp and single LED light condition. Thus, this study indicates that the ratio of blue to red LEDs is important for the morphology, growth, and phenolic compounds with anthocyanin properties in the two lettuce cultivars tested.

• Korean Journal of Soil Science and Fertilizer
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• v.43 no.4
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• pp.490-497
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• 2010

A chitinolytic bacterium having a strong antagonistic activity against various pathogens including Phytophtora capsici was isolated from rhizosphere soil, and identified as Lysobacter enzymogenes (named as LE429) based on 16S rRNA gene sequence analysis. This strain produced a number of substances such as chitinase, ${\beta}-1$, 3-glucanase, lipase, protease, gelatinase and an antibiotic compound. This antibiotic compound was purified by diaion HP-20, sephadex LH-20 column chromatography and HPLC. The purified compound was identified as phenylacetic acid by gas chromatography-electron ionization (GC-EI) and gas chromatography-chemical ionization (GC-CI) mass spectrometry. In field experiment, pepper plants were treated by the strain LE429 culture (CB), neem oil solution (NO), combination (CB+NO) or control (CON). Plant height and number of branches, flowers and pods of pepper plant in CB treatment were generally highest, and followed by CB+NO, CON and NO. The fungal pathogens were strongly inhibited, while several insect pests were discovered in CB treatment. Any insect pests were not found, while all fungal pathogens tested were not suppressed in NO treatment. However, in CB+NO treatment, non incidence of fungal pathogens and insect pests were found. The strain LE429 producing secondary metabolites with neem oil should be a potential agent to control fungal diseases and insect pests.

• Clean Technology
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• v.4 no.1
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• pp.45-59
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• 1998

Continuous immobilized-cell culture was carried out for the production of kasugamycin, a secondary metabolite by a filamentous bacteria, Streptomyces kasugaensis, with an intention of reducing waste generation. A sporulation medium was developed for production of bulk amounts of spores, and the spores were entrapped into celite biosupports for immobilization. It was possible to effectively keep the immobilized-cells inside the reactor during the continuous culture by an efficient immobilized cell separator of decantor type on the outlet of the fermentor. Using this continuous immobilized-cell fermentor system, we investigated the effects of feed substrate and phosphate concentrations on kasugamycin production and chemical oxygen demand(COD). Comparing with the conventional suspended-cell batch culture, the kasugamycin productivity was observed to increase by 2.5 times, whereas COD per unit kasugamycin production decreased by 2.3 times in the continuous immobilized-cell culture. Based on these results, the continuous immobilized-cell system was considered to be a cleaner bioprocess than the conventional batch suspended-cell system.

• Journal of Life Science
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• v.17 no.11
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• pp.1517-1522
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• 2007

Gap junction channels formed by two adjacent cells allow the passage of small molecules up to ${\sim}\;1\;kDa$ between them. Hemichannel (connexon or half of gap junction) also behaves as a membrane channel like sodium or potassium channels in a single cell membrane. Among 26 types of connexin (Cx), $Cx32^*43E1$ (a chimera in which the first extracellular loop of Cx32 has been replaced with that of Cx43), Cx38, Cx46, and Cx50 form functional hemichannels as well as gap junction channels. Although it is known that Xenopus oocytes express endogenous connexin 38 (Cx38), its biophysical characteristics at single channel level are poorly understood. In this study, we performed single channel recordings from single Xenopus oocytes to acquire the biophysical properties of Cx38 including voltage-dependent gating and permeation (conductance and selectivity). The voltage-dependent fast and slow gatings of Cx38 hemichannel are distinct. Fast gating events occur at positive potentials and their open probabilities are low. In contrast, slow gatings dominate at negative potentials with high open probabilites. Based on hi-ionic experiments, Cx38 hemichannel is anion-selective. It will be interesting to test whether charged amino acid residues in the amino terminus of Cx38 are responsible for voltage gatings and permeation.

• KSBB Journal
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• v.22 no.5
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• pp.356-361
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• 2007

Cyanobacteria are a very old group of prokaryotic organisms that produce very diverse secondary metabolites, especially non-ribosomal peptide and polyketide structures. Although some cyanobacteria produce lethal toxins such as microcystins and anatoxins, some may be useful either for development into commercial drugs or as biochemical tools. Detection of unknown secondary metabolites was carried in the present study by a screening of 98 cyanobacterial strains from Cyanobiotech GmbH in order to establish a screening process, isolate pure substances and determine their bioactivities. A degenerated polymerase chain reaction technique as molecular approaches has been used for general screening of NRPS gene and PKS gene in cyanobacteria. A putative PKS gene was detected by DKF/DKR primer in 38 strains (38.8%) and PCR amplicons resulted from a presence of NRPS gene were showed by MTF2/MTR2 primer in 30 strains (30.6%), respectively. A screening of interesting strains was performed by comparing PCR screening results with HPLC analyses of extracts. HPLC analysis for a detection of natural products was performed in extracts from biomass. 5 strains were screened for further scale-up processing. 7 pure substances were isolated from the scale-up cultures and tested for bioactivities under consideration to purity, amount and molecular weight of substances. One substance isolated from CBT 635 showed cytotoxic activity. This substance may be regarded as Microcystin LR.

• Horticultural Science & Technology
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• v.30 no.5
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• pp.509-518
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• 2012

The objective of this study was to determine the effect of heat shock treatments on the phytochemicals including antioxidants and anticancer materials in kale (Brassica oleracea L. var. acephala) sprouts. In study I, kale sprouts grown under the growing system for four days were soaked at 40, 50, or $60^{\circ}C$ distilled water for 10, 30, or 60 seconds, and in study II, kale sprouts were soaked at $50^{\circ}C$ distilled water for 10, 20, 30, 45, or 60 seconds. After the heat shock treatments, the sprouts were transferred into normal growing conditions and recovered there for two days. Fresh and dry weights, electrolyte leakage, total phenolic concentration, antioxidant capacity, total flavonoid concentration, phenylalanine ammonia-lyase (PAL) activity, and glucosinolates content of the sprouts were measured before and after the heat shock treatments. As a result, there was a significant decrease in the fresh and dry weight of kale sprouts treated with heat shock compared with control at harvest in study I. Especially, heat shock at $60^{\circ}C$ lead to more pronounced growth inhibition compared with heat treatments at 40 and $50^{\circ}C$. Electrolyte leakage by cell collapse was the highest in the sprouts exposed to $60^{\circ}C$ distilled water, which agreed with the growth results. Heat shock at $50^{\circ}C$ significantly induced the accumulation of phenolic compounds. In study II, fresh weight of kale sprouts at $50^{\circ}C$ heat shock showed a significant decrease compared with the control at one and two days after the treatment. However, the decrease was minimal and dry weight of kale sprouts was not significantly different from that in control. In contrast, the heat shock-treated kale sprouts had higher level of total phenolic concentration than control at harvest. Heat shock treatments at $50^{\circ}C$ for 20 seconds or more showed at least 1.5 and 1.2 times higher total phenolic concentration and antioxidants capacity than control, respectively. The change of the total flavonoid concentration was similar with that of antioxidants. PAL activity after 24 hours of heat shock was higher in all the heat shock-treated sprouts than that in control suggesting heat shock may stimulate secondary metabolic pathway in kale sprouts. Seven glucosinolates were identified in kale sprouts and soaking the sprouts with $50^{\circ}C$ water for 20 seconds had a pronounced impact on the accumulation of total glucosinolates as well as two major glucosinolates, progoitrin and sinigrin, at harvest. In conclusion, this study suggests that heat shock using hot water would be a potential strategy to improve nutritional quality of kale sprouts by inducing the accumulation of phytochemicals with antioxidant and anticancer properties.

• Journal of Applied Biological Chemistry
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• v.61 no.1
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• pp.83-91
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• 2018

Advances in bacterial and fungal genome mining uncover a plethora of cryptic secondary metabolite biosynthetic gene clusters. Guided by the genome information, targeted transcriptional derepression could be employed to determine the product of a cryptic gene cluster and to explore its biological role. Monascus spp. are food grade filamentous fungi popular in eastern Asia and several genome data belong to them are now available. We achieved transcription activation of a cryptic fungal polyketide synthase-nonribosomal peptide synthase gene Mpfus1 in Monascus purpureus ${\Delta}MpPKS5$ by inserting Aspergillus gpdA promoter at the upstream of Mpfus1 through double crossover gene replacement. The gene cluster with Mpfus1 show a high similarity to those for the biosynthesis of conjugated polyene derivatives with 2-pyrrolidone ring and the mycotoxin fusarin is the representative member of this group. The ${\Delta}MpPKS5$ is incapable of producing azaphilone pigment, providing an excellent background to identify chromogenic and UV-absorbing compounds. Activation of Mpfus1 resulted in a yellow hue on mycelia and its methanol extract exhibit a maximum absorption at 365 nm. HPLC analysis of the organic extracts indicated the presence of a variety of yellow compounds in the extract. This implies that the product of MpFus1 is metabolically or chemically unstable. LC-MS analysis guided us to predict the MpFus1 product and to propose that the Mpfus1-containing gene cluster encode the biosynthesis of a desmethyl analogue of fusarin. This study showcases the genome mining in Monascus and the possibility to unveil new biological activities embedded in it.

• KSBB Journal
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• v.16 no.5
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• pp.516-522
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• 2001

The large-scale production of antibiotics by filamentous mycelial organism requires and adequate supply of dissolved oxygen. In terms of productivity, it means that oxygen transfer is the rate-limiting step. Therefore, the oxygen transfer coefficients(K$\_$L/A) were determined in a broth involving a filamentous mycelial organism such as Streptomyces avermitilis for use in fermentations. To determine (K$\_$L/A) inn a stirred vessel, a great deal of effort is required to provide all the cells with a sufficient oxygen supply. To overcome the oxygen limitation in a batch culture, a fed-batch culture was applied to control the growth rate by an intermittent supply of nutrients. Thus, it was possible to maintain a suitable dissolved oxygen concentration at a low agitation rate. The optimal agitation speed was 350 rpm at low cell concentrations (below 7 g/L) by considering the efficiency of agitation and shear stress. The (K$\_$L/A) was found to decrease from 64.26 to 29.21h.$\^$-1/ when the biomass concentration was increased from 9.82 to 12.06 g/L. In addition, and increase in viscosity was also observed during the growth phase. By comparing the (K$\_$L/A) values for the various agitation and aeration rates, it was found that the effect of an increase in (K$\_$L/A) by aeration was reduced dramatically at high biomass concentrations. However, this effect was not observed when altering the agitation rate. This suggests that controlling the dissolved oxygen concentration by altering the agitation rate was more efficient than increase the aeration rate.

• Korean Journal of Food Science and Technology
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• v.42 no.3
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• pp.257-262
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• 2010

Patulin, a secondary metabolite of mold, is commonly found in rotten apples. Many countries regulate patulin at levels ranging from 30 to $50\;{\mu}g/L$. Most analytical methods for removing patulin from apple juice include liquid-liquid extraction (LLE), which is time and labor intensive. To replace the LLE method, a solid-phase extraction (SPE) method has been developed for apple juice and unfiltered apple juice. A portion of the test sample was applied to a macroporous copolymer cartridge and washed with 5 mL of 1% sodium bicarbonate, followed by 5 mL of 1% acetic acid. Patulin was eluted with 5 mL of 2% acetonitrile in anhydrous ethyl ether. The mobile phase was tetrahydrofuran in water (0.8:99.2) and was detected with a UV detector at 276 nm. Recoveries ranged from 95 to 101% in test samples, and the minimum detectable level was 30 ppb. Because this SPE method is fast, easy, reliable, and inexpensive, it could be applicable for companies or analytical agencies to analyze patulin concentrations in apple juice.

• KSBB Journal
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• v.20 no.4
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• pp.285-290
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• 2005

Benzophenanthridine alkaloids - sanguinarine, chelirubine, macarpine, and chelerythrine are produced from Eschscholtzia californica (Californica Poppy, used as a sedative by Native Americans) and most of them are derived from dihydrosanguinarine. The properties of sanguinarine are the basis of its antimicrobial activity and its use in chemosurgery and skin cancer excision. For overproduction of sanguinarine from E. californica, yeast extract was used as elicitor and the elicited cell's metabolites were checked. Sanguinarine production was increased intracelluarly about 8 times in the cell and 5 times extracelluarly. We have peformed proteomic analysis of proteins sequentially extracted from E. califormica suspended cells which were cultured with elicitor, an increase of spot intensity was seen at 24 hours following elicitation. These proteins were separated by two-dimensional electrophoresis (2-DE). We found several spots that were expected to be related to benzophenanthridine alkaloids production by comparing the production profiles of metabolites such as sanguinarine. These results demonstrate the use of metabolite analysis as a tool for detecting target proteins related to metabolites production pathway.