• Korean Journal of Food Science and Technology
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• v.29 no.1
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• pp.126-132
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• 1997

Calcium carbonate ($CaCO_3$) immobilized with alginate was studied as buffer system to enhance the cultivation efficiency of Bifidobacterium longum (ATCC 15707) which is inhibited at low pH. To test the bufferring effect of the immobilized $CaCO_3$ beads, pH value in each modified trypticase-proteose peptone-yeast (TPY) broth which is adjusted to pH 4.0 with acetic acid, lactic acid and complex solution of acetic and lactic acid, 3:2 (M:M) was tested by concentration of $CaCO_3$ bead and reaction time. The bufferring effect of $CaCO_3$ bead became higher with increasing the amount of $CaCO_3$ bead in the acidic solution. The growth rate of bifidobacteria and bufferring effect were examined in relation to the amount of $CaCO_3$ bead and concentration of glucose in the modified TPY media. The growth rate of bifidobacteria and bufferring effect were increased with increasing the amount of $CaCO_3$ bead and concentration of glucose. Also, the exponential time of bifidobacteria became longer with increasing the amount of $CaCO_3$ bead and concentration of glucose in the modified TPY media. When we observed the growth rate of bifidobacteria by the method of pH-controlled culture and $CaCO_3$ buffer system, the $CaCO_3$ buffer system was more effective than that of pH-controlled culture. Therefore, this $CaCO_3$ buffer system may be useful as a method to enhance of the cultivation efficiency of bifidobacteria.

• Mycobiology
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• v.32 no.1
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• pp.1-5
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• 2004

The present study was carried out to investigate morphological characteristics of pseudosclerotia of Grifola umbellata formed by artificial cultures. Isolate G. umbellata DUM GUS-01 was obtained from sclerotium cultivated in field. The fungal isolate was cultured on PDYM broth, PDYMA(potato dextrose yeast malt agar) and oak sawdust media at $20^{\circ}C$ under the dark condition. G. umbellata DUM GUS-01 showed a volumetric increment of fungal lumps rather than mycelial growth. Particularly, G. umbellata DUM GUS-01 produced a large amount of melanin pigments in all culture treatments. The color of the fungal mass has been changed into grey gradually, and then formed melanized rind-like structure on its superficial part. The fungal structures which were covered with melanized rind-like layer were named as pseudosclerotia of G. umbellata. The pseudosclerotia of G. umbellata DUM GUS-01 formed a new white mycelial mass, which was swollen out of the melanized rind structure for its volumetric increment. When the pseudosclerotia were sectioned, their structure was discriminated from two structures such as a melanized rind-like structure layer formed by aggregation of aged mycelia and a white mycelial mass with high density. As results of scanning electron microscopic examination, the pseudosclerotia of G. umbellata DUM GUS-01 which were formed in in vitro conditions were similar to the sclerotia of G. umbellata cultivated in natural conditions except for the crystals formed in medula layer of natural sclerotia. Although size, solidity of rind structure and mycelial compactness of pseudosclerotia were more poor than those of natural sclerotia, the morphological structure and growth pattern of pseudosclerotia were very similar to those of natural sclerotia. Therefore, it is probable to induce pseudosclerotia to sclerotia of G. umbellata in in vitro conditions. Consequently, it seems that the induced pseudosclerotia can be used as inoculum sources to substitute natural sclerotia in field cultivation.

• Microbiology and Biotechnology Letters
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• v.9 no.2
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• pp.71-75
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• 1981

Studies were made to optimize the simultaneous hydrolysis-fermentation (SSF) process for the production of ethanol from rice straw. Trichoderma sp. KI 7-2 was selected to produced cellulase by solid culture for SSF. Ethanol production was highest when the SSF process utilized koji culture of the fungus grown on a medium of wheat bran-rice straw 3 : 2 mixture with moisture content of 50% adjusted to pH 4.5 for 7 days as the enzyme source. It was found that pretreatment of the substrate is not necessary. To ferment 1g of rice straw by SSF 2.47 units of cellulase were required, and the initial yeast concentration of 2.5$\times$10$^{7}$ cell/$m\ell$ was found to be sufficient. Optimum pH and temperature for the process were 4.5 and 4$0^{\circ}C$, respectively. It was also found that higher ethanol concentration in the broth can be obtained by the addition of substrate or substrate and enzyme to SSF broth.

• Journal of Mushroom
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• v.14 no.4
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• pp.191-196
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• 2016

This study was conducted to investigate optimum conditions for mass production of ntagonistic microbes Alcaligenes sp. HC12. Alcaligenes sp. HC12 had a potent biological control agent to control browning disease caused by Pseudomonas agarici. Alcaligenes sp. HC12 markedly showed the antagonistic activity against Pseudomonas agarici, the most destructive pathogen of cultivated mushrooms. To define the optimum conditions for the mass production of the Alcaligenes sp. HC12, we have investigated optimum culture conditions and effects of various nutrient source on the bacterial growth. The optimum initial pH and temperature were determined as pH 9.0 and $30^{\circ}$, respectively. The optimal concentration of medium elements for the growth of pathogen inhibitor bacterium(Alcaligenes sp. HC12) was determined as follows: 0.5% dextrine, 1.5% yest extract, 1.0% $NaNO_3$, 0.5% $KH_2PO_4$, and 1.5% asparagine.

• Korean Journal of Food Science and Technology
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• v.49 no.6
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• pp.610-616
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• 2017

${\gamma}$-Glutamyltranspeptidase (GGT, EC 2.3.2.2) transfers ${\gamma}$-glutamyl moiety from glutamine to amino acids or peptides and hydrolyzes glutamine to glutamate and ammonia. In order to overproduce ${\gamma}$-glutamyltranspeptidase from Bacillus amyloliquefaciens (BAGGT), the encoding gene was cloned and expressed in Bacillus subtilis. The productivity of BAGGT in Bacillus subtilis was improved by 42-fold by using a dual-promoter system that was generated by combining promoters from B. subtilis ${\alpha}$-amylase and BAGGT genes. Through optimization of medium composition by Plackett-Burman design and central composition design, BAGGT was produced at $18.3{\times}10^7U/L$ of culture in the optimized medium. Compared to previously used Luria-Bertani medium, the optimized culture medium (15 g/L molasses, 60 g/L corn steep liquor, 6 g/L yeast extract, 4 g/L NaCl, 6 g/L $K_2HPO_4$, and 2 g/L $KH_2PO_4$), resulted in a 4.3-fold increase in production of BAGGT.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.915-921
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• 2001

Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals, and reduces the phosphorus pollution of animal waste. In order to express a high level of fungal phytase in Saccharomyces cerevisiae, various expression vectors were constructed with different combinations of promoters, translation enhancers, signal peptides, and terminator. Three different promoters fused to the phytase gene (phyA) from Aspergillus niger were tested: a galactokinase (GAL1) promoter, glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter, and yeast hybrid ADH2-GPD promoter consisting of alcohol dehydrogenase II (ADH2) and a GPD promoter. The signal peptides of phytase, glucose oxidase (GO), and rice amylase 1A(RAmy1A) were included. Plus, the translation enhancers of the ${\Omega}$ sequence and UTR70 from the tobacco mosaic virus (TMV) and spinach, respectively, were also tested. Among the recombinant vectors, pGphyA06 containing the GPD promoter, the ${\Omega}$ sequence, RAmy1A, and GAL7 terminator expressed the highest phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase was also performed by inserting an endoplasmic reticulum (ER) retention signal, KDEL sequence, into the C-terminus of the phytase within the vector pHphyA-6. It appeared that the KDEL sequence directed most of the early expression of phytase into the intracellular compartment yet more than $60\%$ of the total phytase activity was still retained within the cell even after the prolonged (>3 days) incubation of the transformant. However, the intracellular enzyme activity of the transformant without a KDEL sequence was as high as that of the extracellular one, thereby strongly suggesting that the secretion of phytase in S. cerevisiae appeared to be the rate-limiting step for the expression of a large amount of extracellular recombinant phytase, when compared with other yeasts.

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

• Journal of Microbiology and Biotechnology
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• v.28 no.5
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• pp.718-731
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• 2018

The beneficial effects of lactic acid bacteria (LAB) have been intensively investigated in recent decades with special focus on modulation of the host intestinal microbiota. Numerous discoveries of effective probiotics are driven by a significantly increasing demand for dietary supplements. Consequently, technological advances in the large-scale production and lyophilization are needed by probiotic-related industries for producing probiotic LAB for commercial use. Our study had a dual objective, to determine the optimum growth medium composition and to investigate appropriate cryoprotective additives (CPAs) for Lactobacillus salivarius, and compare its responses with other Lactobacillus species. The one-factor-at-a-time method and central composite design were applied to determine the optimal medium composition for L. salivarius cultivation. The following composition of the medium was established (per liter): 21.64 g maltose, 85 g yeast extract, 1.21 ml Tween 80, 6 g sodium acetate, $0.2g\;MgSO_4{\cdot}7H_2O$, $0.02g\;MnSO_4{\cdot}H_2O$, $1g\;K_2HPO_4$, $1.5g\;KH_2PO_4$, $0.01g\;FeSO_4{\cdot}7H_2O$, and 1 g sodium citrate. A cryoprotective additive combination comprising 10% (w/v) skim milk and 10% (w/v) sucrose supplemented with 2.5% (w/v) sodium glutamate was selected for L. salivarius, and its effectiveness was confirmed using culture-independent methods in the freeze-dried cells of the Lactobacillus strains. In conclusion, the optimized medium enhanced the species-specific cultivation of L. salivarius. On the other hand, the cryoprotective effects of the selected CPA mixture may also be dependent on the bacterial strain. This study highlights the necessity for precise and advanced processing techniques for large-scale production of probiotics in the food and feed industries.

• Microbiology and Biotechnology Letters
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• v.30 no.3
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• pp.241-246
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• 2002

Seven lactic acid bacteria showing highly inhibitory activities against Salmonella gallinarum, Staphylococcus aureus, and Escherichia coli were isolated from Korean native chickens. The inhibitory activities were insensitive to various pretenses indicating that the inhibitory substance is not proteinaceous. The culture broths seem to contain other inhibitory substances in addition to lactic acid. The metabolic profile of organic acids produced by Lactobacillus pentosus K34 was investigated by GC-MSD and 28 different organic acids were detected in the culture broth. Compared with the prominent lactic acid, acetic acid, formic acid, the thirdly most abundant phenyllactic acid showed high inhibitory activity against S. gallinarum. After pHs of the acids were adjusted to S, the inhibitory activities of lactic acid, acetic acid, formic acid against S. gallinarum were greatly reduced while the inhibitory activity of phenyllactic acid was unchanged. The inhibitory activity of the phenyllactic acid was specifically high against S. gallinarum and S. aureus but very low against yeast and mold.

• Korean Journal of Microbiology
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• v.51 no.4
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• pp.407-418
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• 2015

This study is focused on establishing the optimal conditions to enhance the production of antilisterial substances by Lactobacillus paracasei BK57 isolated from Baikkimchi. In addition, the growth and in situ lactic acid and bacteriocin production of this strain were investigated during the manufacture of fresh cheese. And then the efficacy of using Lactobacillus starter as a protective culture to improve the safety of fresh cheese against Listeria monocytogenes KCTC 3569 was estimated. Maximum growth rate and activity of antibacterial substances were obtained in Lactobacilli MRS broth at $37^{\circ}C$ with controlled pH 6.0 after 30 h of incubation under aerobic condition. However, the growth rate and antimicrobial activity of bacteriocin produced in whole milk supplemented with yeast extract (2.0%) as a substrate were lower than those obtained in MRS broth. Live cells and cell-free culture supernatant of BK57 strain were effective in the suppression of L. monocytogenes in milk, whereas the inhibitory of the bacteriocin obtained from BK57 strain was higher in BHI broth than in milk. During storage at $4^{\circ}C$ and $15^{\circ}C$ for 6 days, no significant difference was found in the cell viability and antimicrobial activity of BK 57 strain in fresh cheese. In samples held at two temperatures, there was at least a 15% reduction in the numbers of the pathogen in fresh cheese artificially contaminated with approximately $10^5CFU/ml$ of L. monocytogenes within 6 days. Our results demonstrated the usefulness of L. paracasei BK57 having antilisterial activity as a biopreservative in the cheese making process.