• Title/Summary/Keyword: resistant starch 3

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Purification and Characterization of Cyclodextrin Glucanotransferase from Paenibacillus sp. JK-12

  • Kang, Yong;Kim, Sung-Koo;Jun, Hong-Ki
    • Preventive Nutrition and Food Science
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    • v.7 no.3
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    • pp.310-316
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    • 2002
  • Extracellular cyclodextrin glucanotransferase (CGTase) from Paenibacillus sp. JK-12 was purified through sev-eral purification steps consisting of ammonium sulfate precipitation and chromatographies on DEAE-sephadex A-50 and Mono QIM HR5/5. The purified CGTase exhibited a single band on SDS-PAGE and was estimated to be approximately 82 kDa. The isoelectric point of the enzyme was 7.2 as determined by isoelectric focusing. The CGTase from Paenibacillus sp. JK-12 had a transglucosylation activity at the C-2 position of L-ascorbic acid. The optimum pH and temperature for the CGTase activity were 8.0 and 5$0^{\circ}C$, respectively. The enzyme activity was stable from pH 6.0 to 9.() and at temperatures up to 55$^{\circ}C$ at pB 8.0, having 80% residual activity. The activity of the CGTase was strongly resistant to metals such as A $g^{+}$ and $Ba^{2+}$ but slightly inhibited by H $g^{+}$, N $i^{2+}$ and $Mg^{2+}$. The enzymeproduced $\alpha$ -cyclodextrin ($\alpha$-CD) and $\beta$-CD as the main products from starch, but not ${\gamma}$-CD.X>-CD.

Development of a Mixed-culture Fermentation Process and Characterization for New Oligosaccharides and Dextran Using Lipomyces starkeyi and Leuconostoc mesenteroides (Lipomyces starkeyi와 Leuconostoc mesenteroides의 혼합배양에 의한 올리고당과 Dextran의 생성 및 생성당의 특성 연구)

  • 허수진;김도만;이인수;장판식
    • Microbiology and Biotechnology Letters
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    • v.27 no.4
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    • pp.304-310
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    • 1999
  • We have developed a new process for the production of new structure oligosaccharides using the mixed-culture fementation of Lipomyces starkeyi KSM22 and leuconostoc mesenteroides B-512FMCM.L.starkeyi KSM22 produces a novel DXAMase(an enzyme containing both dextranase and amylase activities). It hydrolyzes the soluble starch and dextran. The hydrolyzates were used as acceptors for dextransucrase of L.mesenteroides to synthesize the new oligosaccharides(NOS). In fermentation, as the concentration of sucrose was increased from 9%(w/v) to 15%(w/v), the yields of dextran(sum of dextran I, MW=66kD, and dextran II, MW=21kD) was increased from 12.7% to 42.5%, and NOS was increased from 3.9% to 5.2% of the theoretical, respectively. The NOS of dp(degree of polymerization) 5 and over was increased from 33.1% to 58.3% of the total NOS. The NOS showed heat resistant up to 12$0^{\circ}C$ and was stable at pHs ranged from 2 to 6. The NOS decreased the pH changes in the culture of S. mutans, and also showed inhibitory effects on the growth of S. aureus or S. typhimurium.

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Genetic Diversity and Thermostabilitical Variants of Corbicula japonica from the Two Main Rivers in Korea (한국의 두 강으로부터 재첩의 유전적 종다양성과 열안정성 변이체)

  • Heo, Man-Gyu;Mun, Du-Ho;Heo, Heung-Uk
    • Journal of Environmental Science International
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    • v.7 no.3
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    • pp.243-250
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    • 1998
  • We examined the genetic variation within the species, the patterns of genetic diversty between populations, thermostability variations of enzymes and temperature tolerances of Corbicula japonica from the two main rivers In Korea. Starch gel electrophoresis was used to examine the genetic variation of 22 locl. Henting experiments of electrophoresis under the condition of 40$\pm$5$^{\circ}$ for 15$\pm$5 min disclose thermostabllity differences, called heat-sensitive and heat-resistant types, within each 디ectrophoretic allozyme. Genetic diversity at the natural species level was high (77.3%), whereas the extent of heat-treat groups was relatively low (52.6%). The genetic diversity trends to decrease from the source of two main siderable high genetic diversity compared with a mean value of C. japonica species, It is recommended that several populations of the species in Korea should be preserved.

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Purification and Biochemical Characterization of β-agarase Produced by Marine Microorganism Cellulophga sp. J9-3 (해양미생물 Cellulophga sp. J9-3이 생산하는 베타-아가레이즈의 분리 및 생화학적 특성)

  • Kim, Da Som;Kim, Jong-Hee;Chi, Won-Jae
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.329-336
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    • 2021
  • Cellulophga sp. J9-3, is a gram-negative, aerobic marine bacterium belonging to the family Flavobacteriaceae. In addition to cellulose degradability, the J9-3 strain is also capable of hydrolyzing agar in the solid and liquid medium, and the production of agarase in the presence of agarose can be remarkably induced by the bacterium. From the cell culture broth of Cellulophga sp. J9-3, ammonium sulfate precipitation and three kinds of column chromatography were successively performed to purify a specific agarase protein, the AgaJ93. Purified AgaJ93 showed the strongest hydrolyzing activity towards agarose (approximately 22%), and even displayed activity towards starch. AgaJ93 hydrolyzed agarose into neoagarotetraose and neoagarohexaose via various oligosaccharide intermediates, indicating that AgaJ93 is an endo-type β-agarase. AgaJ93 showed maximum activity at a pH of 7.0 and temperature of 35 ℃. Its activity increased by more than six times in the presence of Co2+ ions. The N-terminal sequence of AgaJ93 showed 82% homology with the heat-resistant endo-type β-agarase Aga2 of Cellulophaga sp. W5C. However, the biochemical properties of the two enzymes were different. Therefore, AgaJ93 is expected to be a novel agarose, different from the previously reported β-agarases.

Glycemic Index Lowering Effects of Breads Supplemented with Resistant Starch, Whole Rye Grain and Fructooligosaccharide (저항전분, 통호밀 및 프락토올리고당을 첨가한 식빵의 개발과 Glycemic index 감소 효과)

  • Park, Min-A;Lee, Joung-Won;Shin, Mal-Shick;Ly, Sun-Yung
    • Korean Journal of Community Nutrition
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    • v.12 no.2
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    • pp.189-197
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    • 2007
  • Low glycemic index (GI) foods have been reported to be very important in the control of blood glucose levels in diseases such as diabetes mellitus and obesity, which are becoming more prevalent in Korea. Bread consumption at breakfast and as a snack is also on the rise. To provide low GI alternatives, breads supplemented with dietary fibers-fructooligosaccharide (FOS), ground whole rye, and 2 types of resistance starch (RS2 and RS4)-were developed. The GIs for these breads were evaluated with 13 healthy college students (M6, F7) and sensory tests were done. Four kinds of breads were made through the modification of a basic recipe for white bread (the control, B) as follows: for Bf, the entire amount of sugar was replaced with FOS. For Ryef, 50% of the white wheat flower was replaced with roughly ground wholerye (20 mesh: 12-20 mesh = 1 : 3) in addition to replacing sugar with FOS. For RS2f and RS4f, 20% of the white wheat flower was replaced with RS2 or RS4, in addition to replacing sugar with FOS. The overall preference score of Bf was similar to that of B, while those of Ryef, RS2f and RS4f were lower than that of B, but showed the acceptable degree of the overall preference. The glycemic indices of Ryef and RS2f were 46.1 and 45.9 respectively, which were significantly lower than the GI of B, 67.8. The GIs of Bf and RS4f were, however, 66.7 and 80.5 respectively, showing no significant difference compared to B. The glycemic loads for a 30 g serving were 9.5, 5.9, 6.2, 11.0 and 9.0 for B, Bf, Ryef, RS2f and RS4f, respectively. In conclusion, addition of RS2 or roughly ground whole rye to the dough formula significantly lowered the GI. Since the preferences shown for those two breads were acceptable, they may be recommended as a substitute for white bread fir persons who need blood glucose management. More studies on the bread making process are required to improve preference and acceptance. Although GI lowering effects for F and RS4 were not found in this study, further studies are needed to verify their effects.

Construction of Amylolytic Industrial Brewing Yeast Strain with High Glutathione Content for Manufacturing Beer with Improved Anti-Staling Capability and Flavor

  • Wang, Jin-Jing;Wang, Zhao-Yue;He, Xiu-Ping;Zhang, Bo-Run
    • 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.

A Gene-Tagging System for Monitoring of Xanthomonas Species

  • Song, Wan-Yeon;Steven W. Hutcheson;Efs;Norman W. Schaad
    • The Plant Pathology Journal
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    • v.15 no.3
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    • pp.137-143
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    • 1999
  • A novel chromosomal gene tagging technique using a specific fragment of the fatty acid desaturase-like open reading frame (des-like ORF) from the tox-argK gene cluster of Pseudomonas syringae pv. phaseolicola was developed to identify Xanthomonas spp.released into the environment as biocontrol agents. X. campestris pv. convolvuli FB-635, a pathogen of Convolvulus arvensis L., (bindweed), was chosen as the organism in which to develop and test the system. A 0.52 kb DES fragment amplified from P. syringae pv. phaseolicola C-199 was inserted into pGX15, a cosmid clone containing a 10.3 kb Eco RI-HindIII fragment derived from the xanthomonadin biosynthetic gene cluster contained in plasmid pIG102, to create a pigG::DES insertion. The 10.8 kb EcoRI-BamHI fragment carrying the pigG:: DES insertion was cloned into pLAFR3 to generate pLXP22. pLXP22 was then conjugated into X. campestris pv. convolvuli FB-635 and the pigG::DES insertion integrated into the bacterial chromosome by marker exchange. Rifampicin resistant, tetracycline sensitive, starch hydrolyzing, white colonies were used to differentiate the marked strain from yellow pigmented wild-type ones. PCR primers specific for the unique DES fragment were used for direct detection of the marked strain. Result showed the marked strain could be detected at very low levels even in the presence of high levels of other closely related or competitive bacteria. This PCR-based DES-tagging system provides a rapid and specific tool for directly monitoring the dispersal and persistence of Xanthomonas spp.released into the environment.

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Characters of motile aeromonads isolated from imported ornamental fish (수입 관상어에서 분리한 motile aeromonads의 특성)

  • Jin, Se-yun;Ko, Chang-yong;Lee, Ye-ji;Jung, Yun-hee;Ju, Seong-cheol;Kim, Eunheui
    • Journal of fish pathology
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    • v.30 no.2
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    • pp.89-96
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    • 2017
  • The majority of freshwater ornamental fish are imported and distributed domestically, causing high risk of exposure to exotic pathogens and drug resistant bacteria in Korea. Aeromonas hydrophila is known as a common species of fresh water bacteria and opportunistic fish pathogen, as well as a species causing zoonotic infection. In this study, we isolated motile aeromonads from various imported freshwater ornamental fish and studied the characters of the isolates. Imported freshwater ornamental fish were purchased on day 1 after the fish were deposited in the aquarium. Bacteria were isolated from the liver, kidney and spleen of fish using 0.5% NaCl containing tryptic soy agar medium. Bacteria were grouped on the basis of their morphological characteristics. The colonies with clear zone on starch-ampicillin agar (SA agar) were tentatively identified as Aeromonas spp. Two hundred and twenty-six strains, about 70% of total isolates were assumed to be Aeromonas spp. Nine isolates were further identified based on the result of the API 20E test and PCR using primers specific for A. hydrophila 16S rRNA gene. The isolates were identified as A. hydrophila and the API 20E test showed differences in trisodium citrate, D-sucrose, D-melibiose, amygdalin and L-arabinose availability between the nine isolates and standard A. hydrophila. The susceptibilities of the isolated bacteria to 10 antibacterial agents were confirmed by the disk diffusion method. Isolated strains were found to be resistant to amoxicillin and ampicillin and sensitive to florfenicol. However, 7 isolates showed multiple drug resistances to erythromycin, oxytetracycline, nalidixic acid etc. Pathogenicity of the isolates was determined by the artificial challenge test on goldfish (Carassius auratus). Three isolates caused 60 ~ 80% mortality in goldfish within 5 days after the initiation of challenge. These results indicate that multiple drug resistant, highly pathogenic and exotic A. hydrophila can spread to domestic aquarium and the preventive treatment of fish before sale is necessary.

The Effects of CO2 Injection and Barrel Temperatures on the Physiochemical and Antioxidant Properties of Extruded Cereals

  • Thin, Thazin;Myat, Lin;Ryu, Gi-Hyung
    • Preventive Nutrition and Food Science
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    • v.21 no.3
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    • pp.271-280
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    • 2016
  • The effects of $CO_2$ injection and barrel temperatures on the physiochemical and antioxidant properties of extruded cereals (sorghum, barley, oats, and millet) were studied. Extrusion was carried out using a twin-screw extruder at different barrel temperatures (80, 110, and $140^{\circ}C$), $CO_2$ injection (0 and 500 mL/min), screw speed of 200 rpm, and moisture content of 25%. Extrusion significantly increased the total flavonoid content (TFC) of extruded oats, and ${\beta}$-glucan and protein digestibility (PD) of extruded barley and oats. In contrast, there were significant reductions in 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, PD of extruded sorghum and millet, as well as resistant starch (RS) of extruded sorghum and barley, and total phenolic content (TPC) of all extrudates, except extruded millet. At a barrel temperature of $140^{\circ}C$, TPC in extruded barley was significantly increased, and there was also an increase in DPPH and PD in extruded millet with or without $CO_2$ injection. In contrast, at a barrel temperature of $140^{\circ}C$, the TPC of extruded sorghum decreased, TFC of extruded oats decreased, and at a barrel temperature of $110^{\circ}C$, PD of extruded sorghum without $CO_2$ decreased. Some physical properties [expansion ratio (ER), specific length, piece density, color, and water absorption index] of the extrudates were significantly affected by the increase in barrel temperature. The $CO_2$ injection significantly affected some physical properties (ER, specific length, piece density, water solubility index, and water absorption index), TPC, DPPH, ${\beta}$-glucan, and PD. In conclusion, extruded barley and millet had higher potential for making value added cereal-based foods than the other cereals.

Distribution and Properties of Microorganisms in Soil of Representative Vegetation of Mt. Nam (남산 주요 식생의 토양 미생물의 분포 및 생리적 특성)

  • 성치남;백근식;김종홍;전영문;김정근
    • The Korean Journal of Ecology
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    • v.21 no.5_3
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    • pp.703-712
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    • 1998
  • Physicochemical factors, microbial population size and the properties of the bacterial isolates were estimated to find out the nature of soil ecosystem of Mt. Nam. Samples were obtained from the surface layer of soils on which specific plant community is developed. Average content of moisture and organic matter of the soils were 21.6% and 17.3%, respectively. These values were similar to those of developing forest soils, but were slightly lower than those of climax ecosystem such as Piagol in Mt. Chiri. Chiri. Content of phosphate was higher than those of other forest soils. The population size of soil bacteria ranged from 27.4 to 195.8 ${\times}\;10^5$ CFU/g. duy soil, and the size is somewhat dependent on the moisture and oranic matter content of soils. A large number of bacteria were able to decompose macromolecules such as starch, elastin and gelatin. Bacterial species composition of each soil was comparatively simple. Pseudomonas, Agrobacterium, Flavobacterium and Xanthomonas which are Gram-negative short rods were widely distributed in the forest soils. The endospore forming Bacillus species were also the main constituents of the soil microflroa. Actinomycetes were widely distributed in the forest soils, but the distribution pattern varied in each site. Most of the actinomycetes were also able to decompose organic macromolecules. The rate of resistant actinomycete strains to antibiotics and heavy metals were lower than those from cultivated soils, but higher than those from well-preserved forest soils. Antibiosis pattern of the actinomycete isolates was similiar to the resistance pattern. This means the forest soils of Mt. nam was somewhat interferred by artificial behabiour.

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