• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1539-1545
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• 2010

In beer, glutathione works as the main antioxidant compound, which also correlates with the stability of the beer flavor. In addition, high residual sugars in beer contribute to major nonvolatile components, which are reflected in a high caloric content. Therefore, in this study, the Saccharomyces cerevisiae GSH1 gene encoding glutamylcysteine synthetase and the Saccharomycopsis fibuligera ALP1 gene encoding ${\alpha}$-amylase were coexpressed in industrial brewing yeast strain Y31 targeting the ${\alpha}$-acetolactate synthase (AHAS) gene (ILV2) and alcohol dehydrogenase gene (ADH2), resulting in the new recombinant strain TY3. The glutathione content in the fermentation broth of TY3 increased to 43.83 mg/l as compared with 33.34 mg/l in the fermentation broth of Y31. The recombinant strain showed a high ${\alpha}$-amylase activity and utilized more than 46% of the starch as the sole carbon source after 5 days. European Brewery Convention tube fermentation tests comparing the fermentation broths of TY3 and Y31 showed that the flavor stability index for TY3 was 1.3-fold higher, whereas its residual sugar concentration was 76.8% lower. Owing to the interruption of the ILV2 gene and ADH2 gene, the contents of diacetyl and acetaldehyde as off-flavor compounds were reduced by 56.93% and 31.25%, respectively, when compared with the contents in the Y31 fermentation broth. In addition, since no drug-resistant genes were introduced to the new recombinant strain, it should be more suitable for use in the beer industry, owing to its better flavor stability and other beneficial characteristics.

• Microbiology and Biotechnology Letters
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• v.15 no.6
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• pp.408-413
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• 1987

Microorganisms capable of degrading the raw naked barley were isolated from soil, and the amylase productivity of each strain was examined on plate contained 2％ raw naked barley. Of the fungi and actinomycetes tested, 71 strains were subjected to subsequent testing for amylase production, and 4 strains were selected as potent amylase producers. Among them, Strain No. 281 produced the most potent raw naked barley saccharifying enzyme, and was identified as genus Rhizopus from morphological and physiological studies. The ratio of raw starch saccharifying activity (RDA) of the crude enzyme derived from the Rhizopus sp. No. 281 was showed 2-3 fold higher than that of commercial enzyme when the raw naked barley was used as the substrate. In the case of uncooked alcohol fermentation using Rhizopus sp. No. 281 glucoamylase preparation, the alcohol yield of the broth was 2％ higher than that of the commercial enzyme.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.5
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• pp.690-700
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• 2012

The experiment was conducted by in vitro fermentation and bacterial community analysis to investigate the reduction of odorous compounds in response to the use of feed additives (FA) during carbohydrate overload in growing pigs. Soluble starch at 1% (control) and various FA at 0.1% Ginseng meal (FA1); Persimmon leaf (FA2); Gingko nut (FA3) and Oregano lippia (FA4) were added to fecal slurry and incubated anaerobically for 12 and 24 h. In vitro parameters and microbial diversity of the dominant bacteria following fermentation were analyzed using Denaturing Gradient Gel Electrophoresis (DGGE), band cloning and sequencing of the V3 region. Results showed that total gas production increased with the advancement of incubation (p<0.05). pH values of FAs and control groups were decreased except the FA4 group which increased somewhat from 12 to 24 h (p<0.05). Ammonia nitrogen ($NH_3$-N) and $H_2S$ gas concentrations were comparatively lower in both stages in FA4 treatment than in the other groups (p<0.05). Hence, $NH_3$-N concentrations in liquid phases were increased (p<0.05) from 12 to 24 h, but the trend was lowest in FA4 than in the other groups at both stages. The total VFA production was comparatively lower and butyrate levels were moderate in FA4 group than in the the other groups during both stages (p<0.05). Indirect odor-reducing compounds such as $NO_2$, $NO_3$ and $SO_4$ concentrations were higher in the FA4 and FA3 than in the other groups at 24 h (p<0.05). After fermentation, ten dominant bands appeared, six of which appeared in all samples and four in only the FA4 treated group. The total number of DGGE bands and diversity was higher in the FA4-group compared to other groups. Additionally, similarity indices were lowest (71%) in the FA4, which represented a different bacterial community compared with the other groups. These findings indicate that $NH_3$-N, $H_2S$ and VFA production was minimal, and pH was also better in the FA4 group than in the other groups. Furthermore, the conversion of odor-reducing indirect compounds or their intermediates was higher in the FA4 group in compared to the other groups. FA4 group generated less odorous products and more indirect products by in vitro fermentation at 24 h, and their microbial pattern appeared to differ from that of the other groups. These findings suggest that this particular FA could change the microbial population, which may have a beneficial effect on odor reduction. It is recommended that the oregano lippia may be supplied to growing pigs as FA along with excess carbohydrate sources to reduce the production of odorous compounds.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.1
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• pp.50-57
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• 2015

This study was conducted to investigate the effect of soybean meal (SM) and soluble starch (SS) on biogenic amine production and microbial diversity using in vitro ruminal fermentation. Treatments comprised of incubation of 2 g of mixture (expressed as 10 parts) containing different ratios of SM to SS as: 0:0, 10:0, 7:3, 5:5, 3:7, or 0:10. In vitro ruminal fermentation parameters were determined at 0, 12, 24, and 48 h of incubation while the biogenic amine and microbial diversity were determined at 48 h of incubation. Treatment with highest proportion of SM had higher (p<0.05) gas production than those with higher proportions of SS. Samples with higher proportion of SS resulted in lower pH than those with higher proportion of SM after 48 h of incubation. The largest change in $NH_3$-N concentration from 0 to 48 h was observed on all SM while the smallest was observed on exclusive SS. Similarly, exclusive SS had the lowest $NH_3$-N concentration among all groups after 24 h of incubation. Increasing methane ($CH_4$) concentrations were observed with time, and $CH_4$ concentrations were higher (p<0.05) with greater proportions of SM than SS. Balanced proportion of SM and SS had the highest (p<0.05) total volatile fatty acid (TVFA) while propionate was found highest in higher proportion of SS. Moreover, biogenic amine (BA) was higher (p<0.05) in samples containing greater proportions of SM. Histamines, amine index and total amines were highest in exclusive SM followed in sequence mixtures with increasing proportion of SS (and lowered proportion of SM) at 48 h of incubation. Nine dominant bands were identified by denaturing gradient gel electrophoresis (DGGE) and their identity ranged from 87% to 100% which were mostly isolated from rumen and feces. Bands R2 (uncultured bacterium clone RB-5E1) and R4 (uncultured rumen bacterium clone L7A_C10) bands were found in samples with higher proportions of SM while R3 (uncultured Firmicutes bacterium clone NI_52), R7 (Selenomonas sp. MCB2), R8 (Selenomonas ruminantium gene) and R9 (Selenomonas ruminantium strain LongY6) were found in samples with higher proportions of SS. Different feed ratios affect rumen fermentation in terms of pH, $NH_3$-N, $CH_4$, BA, volatile fatty acid and other metabolite concentrations and microbial diversity. Balanced protein and carbohydrate ratios are needed for rumen fermentation.

• 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.

• Microbiology and Biotechnology Letters
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• v.16 no.6
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• pp.494-498
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• 1988

The optimum conditions for glucoamylase production, and ethanol productivity of the transformant TSD-14 were investigated as compared with the parental strains. The properties of TSD-14 were comparatively similar to the donor S. diastaticus IFO 1046 as regards the conditions of glucoamylase production and ethanol productivity. The soluble starch was the most effective carbon source for the glucoamylase production. While inorganic nitrogen sources did not prompt cell growth and enzyme production, the organic nitrogen sources generally enhanced both cell growth and glucoamylase production. The metal salts such as FeSO$_4$, MgSO$_4$, MnCl$_2$, and NiSO$_4$were favorable to the enzyme production. And the optium temperature and initial pH for glucoamylase production were 3$0^{\circ}C$ and 5. The transformant TSD-14 produced 8.3%(v/v) ethanol from 15% sucrose medium, 4.8%(v/v) ethanol from 15% soluble starch medium, and 7.5%(v/v) ethanol from 15% liquefied potato starch medium. The corresponding fermentation efficiency were 84% , 45% and 70%, respectively.

• Journal of Life Science
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• v.7 no.1
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• pp.30-38
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• 1997

For screening thermostable $\alpha$-amylase from thermophiles, various samples from extreme environments such as hot spring and sewage near them, and compoat, wereexamined microbial growth in enrichment culture medium at 55$\circ$C on the assumption that enzymes from thermophiles are inevitable thermostable. One strain showing higher $\alpha$-amylase activity was pure cultured and designated as Bacillus sp. TR-25 from the results of morphological, cultural and physiological characteristics. The most important carbon sourses for the enzyme production were soluble starch, dextrin, potato starch and corn starch. Glucose and fructose had a catabolite repression on the enzyme production. The good nitrogen sources for the enzyme production were yeat extract, nutrient broth, tryptone, corn steep liquor and ammonium sulfate. The enzyme production was accelerated by addition of CaCl$_{2}$. $\cdot $ H$_{2}$O. The optimal medium composition for the enzyme production was soluble starch 2.0%, yeast extract 0.55, CaCl$_{2}$ $\cdot $ 2H$_{2}$O 0.015, Tween 80 0.001%, pH8.0, respectively. In jar fermenter culture, this strain shows a rapid growth and required cheaper carbon and nitrogen source. These properties are very useful to fermentation industry. The $\alpha$-amylase of this strain demonstrated a maximum activity at 80$\circ$C, pH 5.0, respectively. And calcium ion did not improve thermostability of the enzyme. At 10$0^{\circ}C$, this enzyme has 235 of relative activity. Transformation was carried out by thermophilic Bacillus sp. TR-25 genomic DNA. As a result, the transformant has increased thermostable $\alpha$-amylase activity.

• Journal of Microbiology and Biotechnology
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• v.11 no.4
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• pp.636-643
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• 2001

Bacillus sp. AIR-5, a strain from soil, produced an extracellular maltopentaose-forming amylase from amylose and soluble starch. This bacterium produced 8.9 g/l of maltopentaose from 40 g/l of soluble starch in a batch fermentation and the maltopentaose made up 90 % of the maltooligosaccharides produced (from maltose to maltoheptaose). The culture supernatant was concentrated using a 30 K molecular weight cut-off membrane and purified by DEAE-Cellulose and Sephadex G-150 column chromatographies. The purified protein showed one band on a native-PAGE and its molecular mass was estimated as 250 kDa. The 250-kDa protein was composed of tetramers of a 63-kDa protein. the isoelectric point of the purified protein was pH 6.9, and the optimum temperature for the enzyme activity was $45^{\circ}C$. The enzyme was quickly inactivated above $55^{\circ}C$, and showed a maximum activity at pH 8.5 and over 90% stability between a pH of 6 to 10. The putative N-terminal amino acid sequence of AIR-5 amylase, ATINNGTLMQYFEWYVPNDG, showed a 96% sequence similarity with that of BLA, a general liquefying amylase.

• Mycobiology
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• v.45 no.2
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• pp.90-96
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• 2017

Different endophytes isolated from the seeds of Sophora flavescens were tested for their ability to produce matrine production. Response surface methodology (RSM) was applied to optimize the medium components for the endophytic fungus. Results indicated that endophyte Aspergillus terreus had the ability to produce matrine. The single factor tests demonstrated that potato starch was the best carbon source and the combination of peptone and $NH_4NO_3$ was the optimal nitrogen source for A. terreus. The model of RSM predicted to gain the maximal matrine production at $20.67{\mu}g/L$, when the potato starch was 160.68 g/L, peptone was 24.96 g/L and $NH_4NO_3$ was 2.11 g/L. When cultured in the optimal medium, the matrine yield was an average of $20.63{\pm}0.11{\mu}g/L$, which was consistent with the model prediction. This study offered an alternative source for the matrine production by endophytic fungus fermentation and may have far-reaching prospect and value.

• Journal of Hydrogen and New Energy
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• v.15 no.4
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• pp.309-316
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• 2004

Chlamydomonas reinhardtii UTEX 90 was cultivated with continuous supply of 2% $CO_2$ using TAP media at $25^\circ{C}$ and produced biomass 1.18 g of dry cell weight/L for 4 days. C. reinhardtii algal biomass(CAB) was concentrated to 20 times by volume and converted into hydrogen and organic acids by anaerobic fermentation using Clostridium butyricum. Organic acids in the fermentate of CAB were consecutively used to produce hydrogen by Rhodobacter sphaeroides KD 131 under the light condition. Approximately 52% of starch in the concentrated CAB which had 4-5.8, 24-26 and 6-7 g/L of starch, protein and fat, respectively was degraded by Cl. butyricum at $37^\circ{C}$. During this process, hydrogen and some organic acids, such as formate, acetate, propionate, and butyrate, respectively were produced. Further conversion of the organic acids in anaerobic fermentate of CAB by Rb. sphaeroides KD131 produced hydrogen from the anaerobic fermentate under the illumination of 8 klux using halogen lamp at $30^\circ{C}$. The result showed that hydrogen was evolved by the anaerobic conversion using Cl. butyricum and then by the photosynthetic fermentation using Rb. sphaeroides KD131. It indicated that the two-step conversion process produced the maximum amount of hydrogen from algal biomass which contained carbohydrate, protein, and fat via organic acids.