• Journal of Microbiology and Biotechnology
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• v.21 no.8
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• pp.808-817
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• 2011

Because of its elevated cellulolytic activity, the filamentous fungus Trichoderma harzianum has a considerable potential in biomass hydrolysis applications. Trichoderma harzianum cellobiohydrolase I (ThCBHI), an exoglucanase, is an important enzyme in the process of cellulose degradation. Here, we report an easy single-step ion-exchange chromatographic method for purification of ThCBHI and its initial biophysical and biochemical characterization. The ThCBHI produced by induction with microcrystalline cellulose under submerged fermentation was purified on DEAE-Sephadex A-50 media and its identity was confirmed by mass spectrometry. The ThCBHI biochemical characterization showed that the protein has a molecular mass of 66 kDa and pI of 5.23. As confirmed by smallangle X-ray scattering (SAXS), both full-length ThCBHI and its catalytic core domain (CCD) obtained by digestion with papain are monomeric in solution. Secondary structure analysis of ThCBHI by circular dichroism revealed ${\alpha}$- helices and ${\beta}$-strands contents in the 28% and 38% range, respectively. The intrinsic fluorescence emission maximum of 337 nm was accounted for as different degrees of exposure of ThCBHI tryptophan residues to water. Moreover, ThCBHI displayed maximum activity at pH 5.0 and temperature of $50^{\circ}C$ with specific activities against Avicel and p-nitrophenyl-${\beta}$-D-cellobioside of 1.25 U/mg and 1.53 U/mg, respectively.

• Korean Journal of Food Science and Technology
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• v.13 no.2
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• pp.121-126
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• 1981

An adenine-guanine doubless and $\beta$-alanine requiring mutant, D-1550-40, which had been derived from Brevibacterium ammoniagenes ATCC 6872, produced a copious amount of xanthosine-5'-monophosphate (XMP). The optimum concentration of adenine and guanine for maximal accumulation of XMP was about 75 ml/l and 100 ml/l for growth. Concentrations higher than 100 mg/l of adenine and guanine inhibited cell growth and XMP accumulation strongly. The inhibition, however, could be recovered by adding $100{\mu}g$ of biotin per liter or 0.3% of casamino acids to the culture solution. High concentrations of phosphate and magnesium salts (1.0 to 1.5%(w/v) in media) were found to be indispensable for XMP accumulation, and the presence of manganese in the culture medium stimulated both growth of cells and accumulation of XMP leaving 5'-inosinic acid unaffected. The maximal accumulation of XMP reached to 60.5 mg/l after 4 days of fermentation which had been started with a medium containing 100 mg of adenine-guanine, 5 mg of $MnSO_4{\cdot}H_2O$ and $100{\mu}g$ of biotin per liter. The specific XMP synthesis(mg of XMP/mg of cells) was increased with the increase of the cell growth rate.

• Journal of Life Science
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• v.14 no.2
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• pp.291-295
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• 2004

Rooibos (Aspalathus linearis) (RB) is a leguminous shrub native to the mountainous areas of the northwestern Cape Province in South Africa. RB tea infusions are the fermentation products of its leaves and fine sterns, and known to have a high antioxidative activity due to the presence of flavonoids and phenolic acids. We investigated the effects of RB tea on the alleviation of oxidative stress on cultured rat cortical neurons in a hypoxic model. Measurement of lactate dehydrogenase (LDH) released into culture media revealed that RB increased cell viabilities in both normoxia (6-18%) and hypoxia (2-24%) dose-dependently (10-100 $\mu\textrm{g}$/ml) on 16 days in vitro (3 days after treatment). Visualization of cell morphology by expression of GFP-Hsc70 fusion protein showed that RB (50 $\mu\textrm{g}$/ml) reduced the average vacuolated soma from 55.4$\pm$4.59% (no RB addition) to 40.9$\pm$6.3% (RB addition) on 5 days after hypoxia. Our results proves efficacy of RB in the neuroprotection of hypoxic neurons and extend application for RB into the prevention and/or treatment of neuronal damages.

• Journal of Life Science
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• v.20 no.11
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• pp.1729-1737
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• 2010

This study was conducted to establish the optimal medium condition for the animal probiotic Lactobacillus plantarum by using onion juice. Cell yield and antioxidant activity increased in proportion to high additive levels of onion juice in medium. Onion juice, sucrose and yeast extract were selected as media ingredient factors and the effects of their mixed ratio in medium were evaluated. The full factorial design consisted of 24 experimental runs and was employed to estimate the main effects of the factors and their interactions. Significant positive effects on cell yield and antioxidant activity was shown with yeast extract and onion juice, respectively. Significant interaction was found only between sucrose and yeast extract in antioxidant activity. Finally, we selected an optimal medium that was composed of (g/l) onion juice, 600; sucrose, 15; yeast extract, 5. The efficiency of this optimum medium was estimated by using a 5 l jar fermenter. As a result, the maximum cell yield was $9.7\;{\log}_{10}$ (CFU/ml) at 12 hr. Cell yield at the end of incubation (20 hr) was $8.9\;{\log}_{10}$ (CFU/ml) and it was very similar with the predicted value, $9.0\;{\log}_{10}$ (CFU/ml). Antioxidant activity of culture was maintained at about 60~65% during all incubation time, resulting in a higher-than-predicted activity of 47.1%.

• The Korean Journal of Mycology
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• v.27 no.4 s.91
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• pp.304-311
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• 1999

Batch kinetics during the exo-polysaccharide (EPS) fermentation of Ganoderma lucidum was investigated as a function of different substrates (glucose and starch), substrate concentration $(1{\sim}7%,\;w/v)$ and subculture (3 times). Logistic model for mycelial growth fitted the experimental data better than Monod and two thirds power model. The Luedeking-Pirt equation was adequate to fit the kinetic data of product formation and substrate consumption. The EPS production was strongly non-growth associated, although it was mixed type. The product formation and sustrate consumption by growth associated mechanism decreased as the concentration of glucose increased, while those of the non-growth associated mechanism increased. However, starch medium increased the growth associated and non-growth associated substrate consumption indicating higher availability of substrate. Also, batch culture in starch medium showed the higher specific growth rate and stability during subculture than those in glucose medium. In conclusion, the enhanced EPS production and stability in the subculture was found to be remarkably improved by use of starch as sole carbon source in medium. The maximum mycelium dry weight and EPS production of 9.463 and 10.410 g/l, respectively, were obtained after shake culture of 7 days at $30^{\circ}C$ from the media containing 7% starch.

• Journal of Microbiology and Biotechnology
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• v.24 no.1
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• pp.26-35
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• 2014

Saccharothrix algeriensis NRRL B-24137 produces naturally different dithiolopyrrolone derivatives. The enzymatic activity of pyrrothine N-acyltransferase was determined to be responsible for the transfer of an acyl group from acyl-CoA to pyrrothine core. This activity was also reported to be responsible for the diversity of the dithiolopyrrolone derivatives. Based on this fact, nine dithiolopyrrolone derivatives were produced in vitro via the crude extract of Sa. algeriensis. Three of them have never been obtained before by natural fermentation: acetoacetyl-pyrrothine, hydroxybutyryl-pyrrothine, and dimethyl thiolutin (holomycin). Two acyltransferase activities, acetyltransferase and benzoyltransferase catalyzing the incorporation of linear and cyclic acyl groups to the pyrrothine core, respectively, were biochemically characterized in this crude extract. The first one is responsible for formation of acetyl-pyrrothine and the second for benzoyl-pyrrothine. Both enzymes were sensitive to temperature changes: For example, the loss of acetyltransferase and benzoyltransferase activity was 53% and 80% respectively after pre-incubation of crude extract for 60 min at $20^{\circ}C$. The two enzymes were more active in neutral and basal media (pH 7-10) than in the acidic one (pH 3-6). The optimum temperature and pH of acetyltransferase were $40^{\circ}C$ and 7, with a $K_m$ value of $7.9{\mu}M$ and a $V_{max}$ of $0.63{\mu}M/min$ when acetyl-CoA was used as limited substrate. Benzoyltransferase had a temperature and a pH optimum at $55^{\circ}C$and 9, a $K_m$ value of $14.7{\mu}M$, and a $V_{max}$ of $0.67{\mu}M/min$ when benzoyl-CoA was used as limited substrate.

• Microbiology and Biotechnology Letters
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• v.43 no.4
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• pp.307-313
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• 2015

Unpolished rice (UR) is considered to be a healthy alternative to white rice when coping with chronic diseases. In the present study, the fermented slurry of unpolished rice (FSUR) was evaluated with respect to its antimicrobial activities and biochemical characteristics, including the quantities of sugar, total soluble sugar, organic acids, free amino acids, pH, and physiological activity. The antimicrobial efficiency of FSUR was assessed using the paper disc-agar diffusion method. FSUR exhibited strong antimicrobial activity against six pathogenic bacterial strains (Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, and Yersinia enterocolitica) and two fermentation strains (Gluconacetobacter intermedius and Lodderomyces elongisporus). The antimicrobial activity of FSUR was higher than the commercial antibiotics, carbenicillin ($50{\mu}g/ml$) and tetracycline ($50{\mu}g/ml$) against S. aureus, E. coli, L. monocytogenes, P. aeruginosa, S. typhimurium, Y. enterocolitica, and L. elongisporus. Also FSUR had a high antioxidant activity. The microorganisms were isolated from FSUR using tryptic soy broth and yeast extract-peptone-dextrose agar media. The isolated microorganisms were characterized using physiological and biochemical analyses as well as by 16S rRNA gene sequencing and phylogenic analysis. 16S rRNA gene sequence analysis showed that the isolated microorganisms had a high similarity to G. intermedius, Lactobacillus casei, Lactobacillus plantarum, and Acetobacter peroxydans.

• Korean Journal of Food Science and Technology
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• v.32 no.6
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• pp.1341-1349
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• 2000

The addition of 5% NaCI to standard plate count (SPC) and bromcresol purple (BCP) agar showed the highest viable cell counts for Jeot-kal samples. The use of 15% glycerol as cryoprotectant showed the highest microbial survival rate at both temperatures, $-20^{\circ}C$ and $-170^{\circ}C$, and on both colony count media, SPC and BCP. During the aging, the pH of Bajirak Jogae-Jeot (fermented clam) decreased from 6.8 to 5.0. Crude protein content was 10% for Bajirak Jogae-Jeot and $6{\sim}7%$ for Myeolchi-Jeot (fermented anchovy). Microbial population of Bajirak Jogae-Jeot was $10^9\;CFU/g$ after 4 weeks of aging, but was only $10^{3-5}\;CFU/g$ in the case of Myeolchi-Jeot. The proportion of Gram positive and catalase negative bacteria in Bajirak Jogae-Jeot increased drastically during the 4 weeks of aging, which showed typical lactic bacterial fermentation. After 2 years' storage of Jeot-kal in liquid nitrogen tank, the cell counts of total aerobic or lactic bacteria were decreased, resulting in about 10% survival rate. Microbial floral change of Jeot-kal was also investigated. In the case of Bajirak Jogae-jeot, the ratio of rod to cocci and that of Gram negative to positive increased after liquid nitrogen storage. But, rod to cocci ratio of Myeolchi-jeot decreased after liquid nitrogen storage. The ratio of yeasts decreased in both cases after storage.

• Food Engineering Progress
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• v.14 no.1
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• pp.14-20
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• 2010

The objective of the current study was to determine the cryoprotective effects of trehaolse on lactic acid bacteria in the frozen yoghurt during long-term frozen storage conditions. The frozen yoghurts were prepared using starter culture containing Lactobacillus bulgaricus and Streptococcus thermophilus, as well as trehalose and sorbitol as a cryoprotectant. The viable cell numbers of lactic acid bacteria in frozen yoghurt did not significantly decreased during six weeks frozen storage conditions. The MRS broth, which contains either trehalose or sorbitol, cultured with L. bulgaricus and/or S. thermophilus, and then the cultured medium was kept in the frozen condition for six weeks. The results indicated that lactic acid bacteria viability significantly increased with trehalose addition (2 and 5%) in the media compared to those of control and sorbitol supplement groups. The lactic acid bacteria viability in the yoghurts was examined on the effects of repeated freeze and thaw events. The freeze-thaw resistance of lactic acid bacteria significantly increased with trehalose supplement in the yoghurt. The major volatile aroma compounds (acetaldehyde, acetone, ethanol, diacetyl, and acetoin) in yoghurt were separated and indentified by headspace GC-FID analysis. Distinct flavor components and their ratios are known as important quality factors for yoghurt notes. Trehalose addition to the yoghurt was not influenced these factors during lactic acid fermentation. The results in this study demonstrated that trehalose potentially can be applicable as an effective cryoprotectant for lactic acid bacteria in the frozen yoghurt products.

• Journal of the Korea Convergence Society
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• v.12 no.9
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• pp.185-191
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• 2021

In our study, we evaluated possibility of distillation process introduction for Korean Rubus coreanus balsamic vinegar production development, has a less complicated and more rapid process leading to higher production volumes and cost savings. In order to investigate the applicability of concentrated vinegar fraction (CVF) and distilled vinegar fraction (DVF) obtained from the distillation process as balsamic and distillation vinegar, acidity, pH, and specific gravity (S.G.) were measured for the vinegar fraction obtained from the distillation process. The acidity of final CVF (60% of initial volume) obtained 4 types of vinegar (acidity; 4.42, 4.84, 5.61, and 6.35) were increased mean 38.1%, compared to initial acidities. Especially, acidity and pH of the final CVF was similar to balsamic vinegar. In addition, acidity of DVF also increased and it of the final fraction was 4.41%, 4.95%, 5.88%, and 6.72%, respectively. And it was suggested that the final DVF could be used as distilled vinegar. In this study, it was confirmed that vinegar can be effectively concentrated through the distillation process, and distilled vinegar can be produced at the same time. However, it is thought that the introduction of the distillation process can be decided only after additional biochemical characteristics such as organic acid content and antioxidant activity measurement for each vinegar fraction are analyzed.