• Microbiology and Biotechnology Letters
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• v.44 no.3
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• pp.254-260
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• 2016

Doenjang samples were prepared by inoculation of multiple starters consisting of two Bacillus spp., one yeast, and one fungus. Doenjang A was fermented with Bacillus amyloliquefaciens EMD17, B. amyloliquefaciens MJ1-4, Pichia farinosa SY80, and Rhizopus oryzae. Doenjang B and C were fermented with the same yeast and fungus but different Bacillus strains; namely, B. amyloliquefaciens EMD17 and B. subtilis CH3-5 for doenjang B, and B. amyloliquefaciens MJ1-4 and B. subtilis CH3-5 for doenjang C. Doenjang D was fermented with microorganisms present in rice straw (control). The doenjang samples were spiked with B. cereus ATCC14579 at two different levels, 10⁴ CFU/g doenjang (I) and 10⁷ CFU/g doenjang (II). All eight doenjang samples were fermented for 70 days at 25℃. Growth of B. cereus was inhibited in doenjang A, B, and C, with the bacterial cell count after 70 days being less than the initial 10⁴ CFU/g added, whereas B. cereus was not inhibited in doenjang D. Doenjang B showed the strongest inhibitory activity against B. cereus, with a cell count of less than 10³ CFU/g after 42 days, even when B. cereus was initially added at 10⁷ CFU/g. Some properties of the doenjang samples, such as pH, TA, and amino-type nitrogen content, were similar to those of doenjang fermented with starters only. The results indicate that carefully selected starters can effectively prevent the growth of B. cereus during doenjang fermentation.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.5
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• pp.736-742
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• 2017

Objective: Experiments were conducted to clone the sequence of Wild Argali short palate, lung and nasal epithelium clone 1 (SPLUNC1) cDNA, and to lay the foundation for further study the biological function of Wild Argali SPLUNC1. Methods: The complete sequence of Wild Argali SPLUNC1 cDNA was generated by rapid amplification of cDNA ends. The entire coding sequence was inserted into the pPIC9K vector and expressed in Pichia pastoris (P. pastoris) GS115. The recombinant SPLUNC1 protein was detected by Western blot and purified by $Ni^{2+}$ chelate affinity chromatography. The test of effect of the protein on Mycoplasma ovipneumoniae (MO) was performed with real-time polymerase chain reaction. Results: The Wild Argali SPLUNC1 cDNA was 1,076 bp with an open reading frame of 768 bp, which encoded a 26.49 kDa protein composed of 255 amino acids. Its amino acid sequence shared 98.4%, 96.9%, 94.5%, 90.2%, 80.8%, 78.4%, 78.3%, 72.5%, 72.3%, 68.8% identity with those of SPLUNC1 cDNA from Ovis aries (accession no. NP_001288334.1), Capra hircus (accession no. XP_005688516.1), Pantholops hodgsonii (accession no. XP_005979709.1), Bos taurus (accession no. NP_776851.1), Felis catus (accession no. XP_006929910.1), Homo sapiens (accession no. NP_001230122.1), Sus scrofa (accession no. NP_001005727.1), Chinchilla lanigera (accession no. NP_001269294.1), Mus musculus (accession no. NP_035256.2), and Rattus norvegicus (accession no. NP_742028.1), respectively. The recombinant protein corresponded to the expected molecular mass of 25.47 kDa as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and it was detected in the supernatant of P. pastoris, and it could be purified. The results from the test of inhibition effect of argali recombinant SPLUNC1 protein on MO showed that the product could inhibit MO very well (p<0.01). Conclusion: The amino acid sequence of Wild Argali SPLUNC1 was different from other organisms. The recombinant SPLUNC1 protein has good biological activity.

• Microbiology and Biotechnology Letters
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• v.43 no.1
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• pp.56-64
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• 2015

It has been known for some time to the wine industry that non-Saccharomyces yeasts play an important role in increasing volatile components through the secretion of extracellular enzymes. The objective of this study was to investigate what types of enzymes are produced by 1,007 non-Saccharomyces yeast strains isolated from Korean fermented foods. Among 1,007 yeast strains, the 566, 45 and 401 strains displayed β-glucosidase, glucanase and protease activity, respectively. In addition, the 563 and 610 strains possessed tolerances against cerulenin and TFL, and the 307 strain was tolerant to 15% ethanol. Yeasts producing harmful biogenic amines and hydrogen sulfide were excluded from further study, and eventually 12 yeast strains belonging to the genera Wickerhamomyces, Hanseniaspora, Pichia, Saccharomyces were identified, based on the 26S rRNA gene sequences. Among the 12 strains, the 9 and 5 strains possessed glucose and ethanol tolerance, respectively. Yeasts belonging to the genus Saccharomyces produced more than 8% alcohol, but non-Saccharomyces yeasts produced only 3% alcohol.

• Journal of the Korean Wood Science and Technology
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• v.41 no.6
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• pp.507-514
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• 2013

In this study, oxalic acid pretreatment of empty fruit bunch (EFB) was performed at different pretreatment temperatures. Also, we evaluated oxalic acid recovery from hydrolysate by electrodialysis. The fermentable sugar concentration in hydrolysate was high at more than $20g/{\ell}$, when pretreatment was carried out at $150^{\circ}C$. At the same time, ethanol production was $3.78g/{\ell}$ after 72 h which correspond to the ethanol yield of 0.21 g/g. On the other hydrolysate (160, $170^{\circ}C$), fermentable sugar was not consumed by Pichia stipitis during fermentation. Most of the oxalic acid was recovered and some of the fermentation inhibitors were removed by electrodialysis. For the electrodialysis treated hydrolysate, ethanol production was increased compared to the original hydrolysate. The highest ethanol production was $5.38g/{\ell}$ after 24 h which correspond to the yield of 0.33 g/g. The ethanol production by simultaneous saccharification and fermentation (SSF) under all pretreatment conditions was more than $15g/{\ell}$ after 96 h. The highest ethanol production was $20.54g/{\ell}$, when pretreatment was performed at $170^{\circ}C$. In particular, ethanol production was increased, when electrodialysis treated hydrolysate was used for SSF.

• 한국균학회소식:학술대회논문집
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• 2015.11a
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• pp.11-11
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• 2015

A total of sixty-six samples of Nuruk, a fermention starter used to make the Korean traditional rice wine, Makgeolli, were collected from central and southern regions of Korea in 2013 and 2014. We classified two groups of the Nuruk samples, "commercial" and "home-made", according to the manufacturing procedure and purpose of use. Commercial Nuruks were made in a controlled environment where the temperature and humidity are fixed and the final product is supplied to Makgeolli manufacturers. Home-made Nuruks were made under uncontrolled conditions in the naturally opened environment and were intended for use in the production of small amounts of home-brewed Makgeolli. We obtained more than five hundred isolates including filamentous fungi and yeasts from the Nuruk samples followed by identification of fungal species. Also we stored glycerol stocks of each single isolate at $-70^{\circ}C$. We identified the species of each isolate based on the sequences of ITS regions amplified with two different universal primer pairs. We also performed morphological characterization of the filamentous fungi and yeast species through observations under the microscope. We investigated the major fungal species of commercial and home-made Nuruks by counting the colony forming units (CFU) and analyzing the occurrence tendency of fungal species. While commercial Nuruks contained mostly high CFU of yeasts, home-made Nuruks showed relatively high occurrence of filamentous fungi. One of the representative Nuruk manufacturers used both domestic wheat bran and imported ones, mainly from US, as raw material. Depending on the source of ingredient, the fungal diversity was somewhat different. Another commercial Nuruk sample was collected twice, once in 2013 and again in 2014, and showed different diversity of fungal species in each year. Nuruks obtained from the southern regions of Korea and Jeju island showed high frequency of yeast such as Saccharomycopsis fibuligera and Pichia species as well as unique filamentous fungus, Monascus species. S. fibuligera was easily found in many Nuruk samples with high CFU. The major filamentous fungi were Aspergillus, Lichtheimia, Mucor and Penicillium species. In order to further our understanding of the isolates and their potential industrial applications, we assayed three enzymes, alpha amylase, glucoamylase and acid protease from 140 isolates out of about five hundred isolates and selected about 10 excellent strains with high enzyme activities. With these fungal isolates, we will perform omics analyses including genomics, transcriptomics, metabolic pathway analyses, and metabolomics followed by whole genome sequencing of unique isolates associated with the basic research of Nuruk and that also has applications in the Makgeolli making process.

• Food Science and Preservation
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• v.12 no.1
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• pp.8-16
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• 2005

To develop natural antimicrobial agents for extending the self-life of Kimchi, the effect of botanical antimicrobial agent-citrus products(BAAC) on microorganisms related to Kimchi spoilage was investigated. The inhibitory effect of BAAC on microorganisms related to Kimchi spoilage was increased according to the concentration of BAAC. Antimicrobial activities of BAAC against microoiganisms related to Kimchi spoilage were remarkably high. The effect of BAAC on the cellular membrane function of microorganisms showed the perturbation of cells in the presence of BAAC. Direct isualization of microbial cells by using both transmission md scanning electron microscope showed microbial cell membrane was destroyed by treating with BAAC. It could be confirmed that BAAC completely inhibit the growth of the test strains. The pH of BAAC-added Kimchi was a little higher than that of the control through the fermentation period. Titratable acidify, vitamin C and viable cells in BAAC-added Kimchi were changed more slowly than those in the control. Sensory evaluation did not show any significant difference between $0.01\%$ BAAC-added Kimchi and the control that showed the best palatabilities during fermentation.

• Food Science and Preservation
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• v.24 no.7
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• pp.1043-1051
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• 2017

Wild yeasts were isolated from domestic non-sterilized Makgeolli and their fermentation characteristics were analyzed to select the best fermentation seed culture. A total of 65 yeast strains isolated yeasts from non-sterilized Makgeolli and Nuruk. In order to select fermentable strains, hydrogen sulfide, $CO_2$ production ability, alcohol tolerance and aroma component production ability were analyzed. To screen the aromatic strains of isolates, media containing cerulenin, 5,5,5-trifluor-DL-leucine (TFL) and API ZYM kit were used. There were 36 strains resistance to cerulenin and all strains produced esterase and demonstrated tolerance against TFL. Hydrogen sulfide, which could degrade the quality of the fermented beverage, was not produced in 34 yeast. The correlation between alcohol tolerance of yeast and carbon dioxide production was analyzed by principal component analysis. YM22, YM31, YM32 and YM37 produced a total of 0.14-0.18 g/72 h of $CO_2$ indicating high fermentability. Alcohol tolerance was measured by alcohol concentration. YM32, YM37 yeast had 20% alcohol tolerance. As a result, alcohol and flavor characteristics of wild yeast isolated from non-sterilized Makgeolli were analyzed and it was confirmed that yeast was suitable for the production of alcohol.

• The Korean Journal of Mycology
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• v.41 no.3
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• pp.185-191
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• 2013

Several yeast colonies were isolated from wild flowers collected from East, West and South coast areas of Korea by plating of flower suspensions on the YPD plates containing antibiotics, streptomycin and ampicillin. Polymerase chain reactions (PCR) were performed for the amplification of D1/D2 region of 26S rDNA for those colonies. PCR-amplified nucleotide sequences were compared using BLAST for their identification. As results, 27 yeast strains belonged to 15 species were isolated from wild flowers collected at Donghae, where is located in eastern coast of Korea. Also, 34 strains belonged to 17 species were isolated from wild flowers of Daecheon, where is located in western coast of Korea. In addition, 22 strains belonged to 13 species were isolated from wild flowers collected at Wando, where is located in southern coast of Korea. Among those 45 species isolated from 3 different collection sites, only 4 species including Cryptococcus laurentii, Metschnikowia koreensis, Pseudozyma rugulosa, and Rhodotorula mucilaginosa were found from all 3 different collection sites. And 5 species including Cryptococcus aureus, Cryptococcus flavus, Hanseniaspora uvarum, Pichia guilliermondii, and Rhodosporidium fluviale were overlapped from the at least 2 different collection sites. Other 23 species were found only in a specific collection sites implying that each area has distinctive yeast flora.

• Microbiology and Biotechnology Letters
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• v.38 no.2
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• pp.151-157
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• 2010

Barley malt produces two different $\alpha$-amylase isozymes (AMY1 and AMY2), which share up to 80% of amino acid sequence identity with each other. However, their enzymatic properties differ remarkably. In this study, five chimeric enzymes between AMY1 and 2 were constructed by staggered extension process (StEP) technique, and their enzymatic properties were characterized. According to the results, chimeric AMY-D2, D8, and E12 showed the mixed or intermediate types of calcium-dependent activity between AMY1 and 2. Meanwhile, only AMY-E10 chimera could be significantly inhibited by barley $\alpha$-amylase/subtilisin inhibitor (BASI) protein. Chimera AMY-C6 showed the same calcium-dependency as AMY1, while AMY-E10 was closely similar to AMY2. As a result, it can be proposed that some amino acid residues in the region II, III, and IV of barley $\alpha$-amylases can play very important roles in the interaction with BASI, and those in III, V, VI, and VII may partly affect on the calcium-dependent activity.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.7
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• pp.996-1006
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• 2019

Objective: This study aimed to gain deeper insights into the dynamic changes in spoilage fungi populations during fermentation and the influence of traditional additives on silage quality. Methods: Elephant grass (Pennisetum purpureum) was prepared without any additive (control), and with the addition of 0.5% salt, and 0.5% salt-0.2% sugar mixture. The fungal community was then determined using a classic culturing method and high-throughput sequencing at 0, 5, 15, and 60 days after ensiling. Results: The results showed that the fungal community of elephant grass silage varied significantly between the natural fermentation without any additive and the two additive groups. The diversity and relative abundance of spoilage molds in the control group were much higher than those in the two treatment groups (p<0.05). Three species of yeasts (Candida sp., Pichia sp., Trichosporon sp.) and four spoilage molds (Fusarium sp., Aspergillus sp., Muco sp. and Penicillin sp.) were the predominant fungi in elephant grass during natural fermentation from 0 to 60 days, which were found to be significantly decreased in salt and sugar additive groups (p<0.05). Meanwhile, the diversity and relative abundance of undesirable molds in the 0.5%-salt additive group were the lowest among all groups. Conclusion: Adding salt and sugar, particularly 0.5% salt, is a promising effective approach to reduce the amount of undesirable fungi thus, improving the silage quality of elephant grass in northern Vietnam.