• Korean Journal of Microbiology
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• v.49 no.2
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• pp.200-204
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• 2013

To contruct amylolytic industrial strains of Saccharomyces cerevisiae which produce ethanol efficiently from raw starch, the Bacillus amyloliquefaciens ${\alpha}$-amylase genes (Amy) or Aspergillus awamori glucoamylase genes (GA1) was separately introduced into the ribosomal DNA loci in the chromosomes of the raw starch fermenting-parental strain (ATCC 9763/$YIp{\delta}AGSA{\delta}$), using double 18S rDNA-integration system. Ethanol production after 3 days of fermentation by the strain that produced ethanol most efficiently from raw starch (ATCC 9763/$YIp{\delta}AGSA{\delta}$/YIpAG2rD) among the transformant strains was 1.5-times higher than that by the parental strain. This new strain generated 9.2% (v/v) ethanol (72 g/L) from 20% (w/v) raw corn starch and consumed 75% of the raw starch content during the same period.

• Journal of the East Asian Society of Dietary Life
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• v.23 no.6
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• pp.833-838
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• 2013

In this study, twelve strains of brewing fungi were individually cultivated on wheat extract broth (WEB), potato dextrose broth (PDB) and malt extract broth (MEB) in order to determine the microorganism with good culture characteristics as well as with amylolytic activity. The strain cultured in PDB exhibited mycelia production from 12.6 g/L (Rhizopus oryzae KACC 45714) to a maximum of 48.0 g/L (Aspergillus oryzae KACC 46959), which was 2.3~9.2 times lower than that of the strain cultured in WEB and 1.7~14.6 times lower than that of the strain cultured in MEB. Accorfing the results, We found that the commercial strains of A. oryzae Suwon, CF1001 and CF1003 had a higher dry cell mass than the wild-type strains KACC 46421, 46423, 46424 and 46959. For Rhizopus sp., the acidity levels in WEB, PDB and MEB were 0.12~0.47%, 0.22~1.0% and 0.16~0.68% (equivalent lactic acid concentration) respectively. For A. oryzae, the acidity levels were 0.06~0.11%, 0.03~0.04% and 0.06~0.08% (equivalent lactic acid concentration), respectively. Amylase enzyme from Rhizopus delemar KACC 46419 exhibited an enzyme activity of 0.013 U and 0.019 U in WEB and MEB cultures, respectively. The enzyme activity of the amylase enzyme from A. oryzae was 0.019~0.037, 0.017~0.033 and 0.028~0.046 U in WEB, PDB and MEB cultures, respectively.

• Korean Journal of Microbiology
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• v.7 no.4
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• pp.159-166
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• 1969

It is necessry to develop and strengthen the activity of enzymic source which in low applied for maggerley brewing as an amylolytic and proteolytic starter, recently in this country the active and strong enzymic starter is required for the better brewing and to substitute another starch material for the present wheat flour. In this study, manufacturing method the strong enzymic source have been developed and established with use of raw wheat bran plus fungal strains of Rhizopus sp. and Aspergillus usamii the culture of starter. The results on experimental the activities of enzymic sources (stater) are as following ; 1. Method of making the enzymic source (starter) is to cultivate the strains of Asperguillus orzyae, Asp. kawachii, Asp usamii and Rhizopus sp. in the acid treated raw or heatboiled wheat bran. 2. The saccharogenic pwoer (S.P.) of enzymic source which consisted of raw bran plus fungi and cultured in it is generally stronger than those of heat-boiled bran plus fungi, the strongest power was shown in the culture of Rhizopus plus raw bran, and the next other is in mixture of Asp.usamii and Rhizopus on raw wheat bran. 3. The most strong alpha amylase activity was expressed in the plot of Asp.oryzae on heat-boiled wheat bran, the next was in the culture of Rhizopus nad Aspergillus usamii on raw wheatbran. 4. The most vigourous acidic proteinase activity was expressed in the micture of raw bran plus Asp. usamii and Rhizopus those were independentlu cu;tured before mixing for neutral proteinase activity, it was shown in the mixed culture of Asp. usamii and Rhizopus on raw wheat bran, the msot active alkaline proteinase activity of enzymic source was found in the plot of raw bran material. 5. For poly-preptidase activity in pH 6.5 it is found that the culture of Rhizopus and Asp.usamii on raw bran was most active among them of enzymic sources. 6. Generally, it is concluded that culture of fungi on acid treated raw wheat bran is stronger in its activity than those of heat boiled wheat bran, especially the culture of Rhizopus nad Asp.usamii on raw bran exhibited the most vigorous and non-polarized activity for all aspects, so it is considered to be most desirable enzymic stater in Korean Maggerley brewing and this would be able to substitute brewing material for the present wheat flour because of its strong and wide hand activity of amylolytic and proteolytic action.

• Korean Journal of Microbiology
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• v.44 no.3
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• pp.264-269
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• 2008

To develop an amylolytic industrial yeast strain producing $\beta$-amylase, the BAMY gene encoding Achlya bisexualis $\beta$-amylase was constitutively expressed under the control of the alcohol dehydrogenase gene promoter (ADC1p) in an industrial strain of Saccharomyces cerevisiae. Yeast transformation was carried out by an integration system containing $\delta$-sequences as the recombination site. The integrative cassette devoid of bacterial DNA sequences was constructed that contains the BAMY gene and $\delta$-sequences. Industrial S. cerevisiae transformed with this integrative cassette secreted 45 kDa $\beta$-amylase into the culture medium. The $\beta$-amylase activity of the transformant was approximately 18.5-times higher than that of A. bisexualis. The multi-integrated BAMY genes in the transform ant were stable after 100 generations of growth in nonselective medium. Hydrolysis of soluble starch and various starches with the enzyme released maltose but not glucose or oligosaccharides.

• Microbiology and Biotechnology Letters
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• v.43 no.4
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• pp.378-384
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• 2015

Nuruk is a fermentation agent, which has been used for the production of traditional Korean alcoholic beverages. The objective of this study was to investigate the effects of temperature on nuruk fermentation. One wheat nuruk sample was fermented at $36^{\circ}C$ for 30 days (TN-A) and another at $45^{\circ}C$ for 10 days followed by $36^{\circ}C$ for 20 days (TN-B). The activities of ${\alpha}$-amylase, glucoamylase, and acidic protease, as well as metabolite contents were measured. Initially, the enzymatic activities increased rapidly regardless of the fermentation temperature. After 3 days of fermentation, the enzymatic activities were maintained in TN-A, but gradually decreased in TN-B until the end of fermentation process. Metabolite analysis using $^1H$-NMR showed that the levels of glucose, glycerol, fructose, mannitol, and lactose initially increased quickly and then decreased in TN-A. However, they initially decreased and then were maintained over the fermentation period in TN-B. The contents of glycine, proline, and serine were higher in TN-A than in TN-B. This study suggests that a constant temperature of approximately $36^{\circ}C$ is appropriate for achieving high amylolytic and proteolytic activities in the production of wheat nuruk.

• Journal of the Korean Applied Science and Technology
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• v.26 no.1
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• pp.10-17
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• 2009

This study investigated the effect of kilning condfition on the diastatic power and activities of protease, $\alpha$-amylase, and $\beta$-amylase in rice malt. Common rice (Oryza sativa) was steeped at $30^{\circ}C$ for 50 h, germinated at $30^{\circ}C$ for 7 days, and kilned at $50^{\circ}C$ for 24 h. The moisture content and enzymatic activities were determined under various kilning times. As a result, the moisture content was reduced from 42.1 % to 3.9% after 24 h of kilning at $50^{\circ}C$. The protease activity of rice malt showed lower value than that of barley malt. All enzymatic activities were decreased during the kilning stage. Results indicated that after prolonged kilning at $50^{\circ}C$, the inactivation of hydrolytic enzymes might be occurred. Even though the amylolytic activity of malted rice showed low value, the rice malt shows the potential characteristics as ingredient for the brewing and cereal industries.

• Korean Journal of Microbiology
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• v.24 no.3
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• pp.308-316
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• 1986

Vertical distribution of heterotrophic bacteria and physico-chemical characteristics were measuted in Kum River estuarine sediments. And interrelationship between heterotrophic bacterka and environmental factors was also studied. The type of sediment of Site 1 was silty clay, and sand at Site 2. Annual pH ranges were between 7.1 and 7.7 in the clay type sediment (Site 1) and 6.9-7.2 in the sand type sediment (Site 2). It was shown that organic matter contents were higher in the clay type sediment than those of sand type sediment. Redox potential values of sediments were decreased rapidly with depth at Site 1, but those of Sete 2 showed vertical fluctuation. Nitrogens(ammonia+amino acid-N, nitrate-N, nitrite-N) and phosphate in the clay type sediment showed higher values than those of sand type sediment. Annual distribution of heterotrophic bacteria were ranged $6.71{\times}10^4$ cells/g dry wt. $-2.50{\times}10^6$ cells/g dry wt. In the clay type sediment and $2.67{\times}10^3$ cells/g dry wt. $-1.94{\times}10^6$ cells/g dry wt. in the sand type sediment. Distribution of proteolytic, lipolytic, and amylolytic bacteria were decreased with the depth and the highest density was found in April and the lowest in January. Bacterial populations in sediments were closely correlated with such environmental factors as pH, redox potential, moisture content, organic matter contents, and inorganic nutrients such as nitrite-N and phosphate-P.

• Microbiology and Biotechnology Letters
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• v.44 no.4
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• pp.488-492
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• 2016

Saccharomycopsis fibuligera is an amylolytic yeast that exhibits raw starch-degrading activity. In this study, adaptive laboratory evolution was performed to improve the tolerance of S. fibuligera to lactic acid by prolonged repeated batch fermentation in which the lactic acid concentration was gradually increased. The evolved S. fibuligera strain exhibited a significantly enhanced tolerance to lactic acid at concentrations up to 2.5% (w/v), as assessed by determining its specific growth rate using a plate assay. Scanning electron microscopy revealed an elongated and perforated morphology of the parent strain under lactic acid stress, indicating that its membrane might be more prone to damage caused by lactic acid than that of the evolved strain.

• Journal of Microbiology and Biotechnology
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• v.9 no.1
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• pp.62-69
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• 1999

In order to investigate the critical amino acid residues involved in the catalytic activities of $\beta$-cyclodextrin glucanotransferase ($\beta$-CGTase) excreted by Bacillus firmus var. alkalophilus, the amino acid residues in $\beta$-CGTase were modified by various site-specific amino acid modifying reagents. The cyclizing and amylolytic activities of $\beta$-CGTase were all seriously reduced after treatment with Woodward's reagent K (WRK) modifying aspartic/glutamic acid, N-bromosuccinimde (NBS) modifying tryptophan, and diethylpyrocarbonate (DEPC) modifying histidine residues. The roles of tryptophan and histidine residues in $\beta$-CGTase were further investigated by measuring the protection effect of various substrates during chemical modification, comparing protein mobility in native and affinity polyacrylamide gel electrophoresis containing soluble starch, and comparing the $K_m$ and $V_{max}$ values of native and modified enzymes. Tryptophan residues were identified as affecting substrate-binding ability rather than influencing catalytic activities. On the other hand, histidine residues influenced catalytic ability rather than substrate-binding ability, plus histidine modification had an effect on shifting the optimum pH and pH stability.

• Journal of Microbiology and Biotechnology
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• v.20 no.9
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• pp.1307-1313
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• 2010

An extracellular ${\alpha}$-amylase from Lactococcus lactis IBB500 was purified and characterized. The optimum conditions for the enzyme activity were a pH of 4.5, temperature of $35^{\circ}C$, and enzyme molecular mass of 121 kDa. The genome analysis and a plasmid curing experiment indicated that $amy^+$ genes were located in a plasmid of 30 kb. An analysis of the phylogenetic relationships strongly supported a hypothesis of horizontal gene transfer. A strong homology was found for the peptides with the sequence of ${\alpha}$-amylases from Ralstonia pikettii and Ralstonia solanacearum. The protein with ${\alpha}$-amylase activity purified in this study is the first one described for the Lactococcus lactis species, and this paper is the first report on a Lactococcus lactis strain belonging to the amylolytic lactic acid bacteria (ALAB).