• Title/Summary/Keyword: Starch mutant

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Physicochemical Characteristics of Starches in Rice Cultivars of Diverse Amylose Contents

  • Yoon, Mi-Ra;Chun, A-Reum;Oh, Sea-Kwan;Hong, Ha-Cheol;Choi, Im-Soo;Lee, Jeong-Heui;Cho, Young-Chan;Kim, Yeon-Gyu
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.57 no.3
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    • pp.226-232
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    • 2012
  • Through the sampling four rice cultivars with differing amylose contents, the relationship between the structural and gelatinization properties of endosperm starches was analyzed. These rice varieties exhibited different chain length distribution ratio within the amylopectin cluster as well as varing amylose levels. The proportion of amylopectin short chains of in Goami cutlivars was higher than the other varieties, whereas the Goami 2 which shows amylose extender mutant properties in the endosperm showed the highest proportion of long chains. In X-ray diffraction analysis of rice starches, the Goami 2 variety displayed a B-type pattern whereas the other varieties were all A-type. Among the cultivars with high and normal rice starch levels, those with the higher amylose contents showed distinctly lower swelling. Goami 2 rice was found to have the highest onset and peak gelatinization temperature from the differential scanning calorimetry results. The four rice cultivars under analysis also showed different rates of hydrolysis by amyloglucosidase. These findings suggest that the composition and chemical structure of the starch content is a major determinant of both the gelatinization and functional properties of rice.

Genetically Engineered Yeast by Heterologous Transformation and Intergeneric Two-Step Protoplast Fusion for Ethanol Fermentation

  • Kim, Young-Ho;Lee, Jae-Ran;Seu, Jung-Hwn
    • Journal of Microbiology and Biotechnology
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    • v.3 no.4
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    • pp.232-237
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    • 1993
  • A strain of yeast which can convert starch directly to ethanol was developed by the intergeneric protoplast fusion between Schwanniomyces alluvius possessing $\alpha$ amylase as well as glucoamylase with debranching activity and FSC-14-75 which previously had been formed from a heterologous transformation and subsequent intergeneric protoplast fusion. Fusants were selected on minimal medium after protoplasts of auxotrophic mutant of S. alluvius fused with heat-treated protoplasts of FSC-14-75 in the presence of 30%(w/v) PEG and 20 mM $CaCl_2$. The fusion frequency was in the range of $10^{-6}$ order. All fusants tested were intermediate types of parental strains for carbon compound assimilation, and their cell volumes were approximately 1.1 times larger than FSC-14-75 and 1.8 times larger than S. alluvius. The fusants were unable to sporulate like FSC-14-75, while S. alluvius could sporulate. In flask scale the most promising fusant, FSCSa-R10-6, produced 7.83%(v/v) and 10.17%(v/v) ethanol from 15% and 20% of liquefied potato starch, respectively, indicating that the fermetation efficiency of each case increased 1.2 times and 1.6 times than that of FSC-14-75. The elution pattern on DEAE-cellulose chromatography showed that FSCSa-R10-6 has four distinct amylase peaks of which two peaks originated from S. alluvius and the other two from FSC-14-75. These results suggest that the enhanced fermentation efficiency of the fusant might be due to almost-complemented parental amylases.

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Global Transcriptome-Wide Association Studies (TWAS) Reveal a Gene Regulation Network of Eating and Cooking Quality Traits in Rice

  • Weiguo Zhao;Qiang He;Kyu-Won Kim;Feifei Xu;Thant Zin Maung;Aueangporn Somsri;Min-Young Yoon;Sang-Beom Lee;Seung-Hyun Kim;Joohyun Lee;Soon-Wook Kwon;Gang-Seob Lee;Bhagwat Nawade;Sang-Ho Chu;Wondo Lee;Yoo-Hyun Cho;Chang-Yong Lee;Ill-Min Chung;Jong-Seong Jeon;Yong-Jin Park
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.207-207
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    • 2022
  • Eating and cooking quality (ECQ) is one of the most complex quantitative traits in rice. The understanding of genetic regulation of transcript expression levels attributing to phenotypic variation in ECQ traits is limited. We integrated whole-genome resequencing, transcriptome, and phenotypic variation data from 84 Japonica accessions to build a transcriptome-wide association study (TWAS) based regulatory network. All ECQ traits showed a large phenotypic variation and significant phenotypic correlations among the traits. TWAS analysis identified a total of 285 transcripts significantly associated with six ECQ traits. Genome-wide mapping of ECQ-associated transcripts revealed 66,905 quantitative expression traits (eQTLs), including 21,747 local eQTLs, and 45,158 trans-eQTLs, regulating the expression of 43 genes. The starch synthesis-related genes (SSRGs), starch synthase IV-1 (SSIV-1), starch branching enzyme 1 (SBE1), granule-bound starch synthase 2 (GBSS2), and ADP-glucose pyrophosphorylase small subunit 2a (OsAGPS2a) were found to have eQTLs regulating the expression of ECQ associated transcripts. Further, in co-expression analysis, 130 genes produced at least one network with 22 master regulators. In addition, we developed CRISPR/Cas9-edited glbl mutant lines that confirmed the role of alpha-globulin (glbl) in starch synthesis to validate the co-expression analysis. This study provided novel insights into the genetic regulation of ECQ traits, and transcripts associated with these traits were discovered that could be used in further rice breeding.

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Agronomic Characteristics of A Mutant for Genic Male Sterility-Chalky Endosperm and Its Utilization on $F_1$ Hybrid Breeding System in Rice (벼 유전자적 웅성불임-심백 돌연변이체의 특성과 그를 이용한 일대잡종 육성체계)

  • ;Mun-Hue Heu
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.40 no.6
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    • pp.684-696
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    • 1995
  • A mutant for chalky endosperm and genetic male sterility (GMS) was newly developed in rice. The two characters were found to be controlled by single recessive gene which has pleiotropic effect, indicating that chalky seeds should be GMS seeds in segregating populations. Chalky seeds showed the same shape and size as normal seeds. However, starch composition of central part of endosperm was looser and shape of starch granules was rounder compared with normal endosperm, resulting in significantly lower grain weight, absolute density and grain hardness in chalky grains than in normal ones. Amylose content and gel consistency of chalky grains were much lower and harder, respectively. Male sterile plants showed much shorter plant height, poorer panicle exsertion and lesser panicle number compared with normal plants. Microspore abortion stage in pollen developmental process was observed as before meiosis. Male sterility of the mutant was stable regadless of temperature and day length. A system breeding hybrid rice using this mutant was discussed, comp ring with other systems utilizing cytoplasmic-genic male sterility(CGMS) and environment sensitive GMS(EGMS). Separation of GMS seeds in mixed seed bulks by specific gravity (1.14∼1.16g / cm3) was successful about 85∼90%. But some mixed normal plants were seemed to be easily removed by the apparent difference in growth characters at seedling stage. The highest natural outcrossing rate of this GMS line was as 17.3 % in a plot treated with 2-row pollinator, I-row GMS, and GA3 + cutting of flag leaf + pollen-scattering by rope.

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Mutational and Nutritional IMprovement of Tylosin Production (Tylosin 생성 우수 균주선별과 Tylosin 발효)

  • 이상희;정병철;이계준
    • Microbiology and Biotechnology Letters
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    • v.23 no.6
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    • pp.647-651
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    • 1995
  • A tylosin-hyperproductive mutant of S. fradiae MNU20 was isolated among 3500 strains obtaincd from either MNNG- or UV-treated Streptomyces fradiae NRRL2702. The composition of optimal medium for tylosin production was formulated as followed as: 4 g soluble starch, 1 g glucose, 1 g corn steep liquor, 7.5 ml soy bean oil, 0.2 g KH$_{2}$PO$_{4}$, 1 g Na$_{2}$S$_{2}$O$_{3}$$\cdot $5H$_{2}$O, 2 g CaCO$_{3}$, 2 g NaCl, 0.001 g CoCl$_{2}$$\cdot $6H$_{2}$O in 1 liter of distilled water. With the optimal medium, S. fradiae MNU20 was able to produce 159 mg tylosin (g biomass)$^{-1}$, indicating that tylosin productivity of Streptomyces fradiae NRRL2702 was increased 14 times higher by mutation. When the effect of valine, succinate, and natural zeolite on tylosin production was investigated by using the optimal medium, these substances essentially enhanced tylosin production and their addition time during culture period appeared to be critical for the increase of tylosin production.

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Production of Gentamicin by Micromonospora purpurea (Micromonospora purpurea에 의한 gentamicin생성)

  • 이묘재;유두영
    • Korean Journal of Microbiology
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    • v.17 no.3
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    • pp.152-159
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    • 1979
  • Using Micromonospora strain, gentamicin was produced by fermentation. The increase in gentamicin productivity was studied by strain improvement and systematic optimization of fermentation process variables. The productivity of parent strain of M.purpurea (ATCC15835) was improved by selection of superior mutant after U.V. irradiation and induction of genetamicin resistance. Potato starch and soy bean meal were the best carbon and nitrogen sources for gentamicin fermentation, respectively. The optimum stimulating concentration of Co ion for gentamicin production was 0.006g $CoCl_2$ per liter of broth. Oxygen ws found to be an important factor for gentamicin yield. The optimum pH for the cell growth and gentamicin production were 7.2 and 6.8 respectively. By controlling the pH, oxygen, and other conditions found in this study at the optimal conditions for cell growth and gentamicin production, the total productivity of gentamicin was increased significantly.

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Amylase Production by Continuous Cultures of Aspergillus oryzae and its Mutants (Aspergllus oryza와 그 변이주의 연적배양에 의한 amylase 생성에 관한 연구)

  • Han, Hong-Eui
    • Korean Journal of Microbiology
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    • v.15 no.2
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    • pp.63-76
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    • 1977
  • Irradiation with high doses of gamma rays induced the reduction of mycelial weight and anaylase activity, and increased relative amylase activity in surface cultures. Biphase in growth curves was shown in aeration-agitation cultures but the behavior of the first phase of growth could be eliminated by replacing the amylasehydrolysed starch substrates, so that enzyme production was shortened ca. 40 hours and relative amylase activity was increased about 3 times higher before onset of autolysis. In the effect of gibberellin on amylase production, the positive stimulation was appeared to only surface culturs of the liquid medium and the negative effect to shake-cultures in a mutant. Trials of various continuous culture were resulted not only the approalch to the value of amylase activity in surface cultures of liquid medium, but also higher productivity than in batch cultures. The culture-degeneration was observed in two-stage continuous culture, but did not appear in continuous elevation culture.

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Characterization of Brewing Yeast Expressing Glucoamylase Selected by Rare Mating. (Rare Mating에 의한 양초효모에서의 glucoamylase 발현 균주 HCS 선별 및 특성)

  • 최병주;장금일;김광엽
    • Microbiology and Biotechnology Letters
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    • v.29 no.4
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    • pp.212-220
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    • 2001
  • Rare mating was used to select a respiratory deficient mutant of Saccharomyces cerevisiae HDC52 strain. Glucoamylase gene of S. diastaticus K114 was developed into the RD mutant which could uptake maximum amount of non-fermentable sugars through the expression of glu- coamplyase gene and the fermentation characteristics of the developed strain HCS were investigated. The size of HCS yeast and HBD52 yeast strain were 13 $\mu\textrm{m}$ and 10$\mu\textrm{m}$ respectively. HCS strain which can uptake maximum amount of non-fermentable sugar through the expression of glucoamylase gene was developed. By karyotype anal- ysis. HCS stain but not RD mutant HBC52 showed a band of 1150 kb chromosome DNA This band should include glcoamylase gene from Saccharomyces diataticus K114 THis strain has glucoamylase which can degrade starch By transduction and contrnuance of glucoamylase gene HCS strain gegraded strach and formed halo. Also, HCS strain maintained the character after 50 generations. Glucoamylase activities of Saccharomyces diastaticus K114 and HCS yeast strains are 9.5 and 2.7~3.4(unit/ml) HCS and HBC52 strain showed similar sugar fermentation patterns and low flocculation In spore and film forming test, HCS and HBC52 strain formed neither spores nor films. In the limit fermentation test, HBC52 strain showed fermentation level of 68% and HCS strain showed 76~78% As the limit attenuation of HBC52 and HCS were ($2.00^{\circ}$P) and ($0.7~0.93^{\circ}$P) This study demon- strates and HCS strain may be used for low carbohydrate beer fermentation.

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Rice Cultivar Breeding for Rice Flour with Low Amylose by Tissue Culture (저아밀로스 특성 및 쌀가루 적합 벼 품종 육성)

  • Park, Jae-Ryoung;Kim, Eun-Gyeong;Jang, Yoon-Hee;Kim, Kyung-Min
    • Journal of Life Science
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    • v.32 no.9
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    • pp.698-705
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    • 2022
  • Rice consumption in Korea is continuously decreasing; therefore, methods for reducing rice stocks are now being proposed. Increasing the consumption of rice and reduce these stocks will require that the rice processing industry be developed beyond simply using cooked rice as food. The Korean people's eating habits have also diversified recently, and the consumption of bread is increasing. Therefore, breeding a cultivar for rice flour is now necessary. Here, a line suitable for processing into rice flour was cultivated by deriving a mutant line by tissue culture of Ilpum. Mutant populations were investigated for major agricultural traits in the field, and grain characteristics and endosperm characteristics were investigated after harvest. Among the lines, line 29111 had similar agricultural characteristics to the background Ilpum, and excellent grain quality, and was selected for further analysis. The physicochemical properties that determine taste were similar or improved. However, unlike Ilpum, 29111 had a non-uniform starch structure and a characteristic powdery endosperm because large and male starch grains are evenly distributed. Therefore, 29111 was viewed as a breeding line suitable for processing and for excellent agricultural characteristics and taste. The added value can be increased by diversifying the uses of rice. The taste was improved and a method for breeding cultivars with grain characteristics for glutinous rice that would satisfy both producers and consumers was presented.

Isolation, Production, and Characterization of Protease from Bacillus subtilis IB No. 11

  • Lee, Min-Hyang;Lee, Kang-Moon;Choi, Yong-Jin;Baek, Yeon-Soo
    • Journal of Animal Science and Technology
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    • v.51 no.6
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    • pp.527-536
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    • 2009
  • A potent protein degrading bacterium was isolated from soil samples of different environments. Polyphasic taxonomic studies and phylogenetic 16S rRNA sequence analyses led to identify the isolate IB No. 11 as a strain of Bacillus subtilis. The isolated strain was recognized to produce protease constitutively, and the maximum production (1.64 units/ml) was attained in a shake flask culture when the isolate was grown at $40^{\circ}C$, for 32 h in basal medium supplemented with starch (0.25%) and gelatin (1.25%) as sole carbon and nitrogen source, respectively. The optimum pH and temperature for the protease activity were determined to be pH 7.0 and $50^{\circ}C$, respectively. $Ca^{2+}$ and $Mn^{2+}$ enhanced remarkably the protease activity but neither showed positive effect on the protease's thermal stability. In addition, it was observed that the protease was fairly stable in the pH range of 6.5-8.0 and at temperatures below $50^{\circ}C$, and it could be a good candidate for an animal feed additive. The inhibition profile of the protease by various inhibitors indicated that the enzyme is a member of serine-proteases. A combination of UV irradiation and NTG mutagenesis allowed to develop a protease hyper-producing mutant strain coded as IB No. 11-4. This mutant strain produced approximately 3.23-fold higher protease activity (6.74 units/mg) than the parent strain IB No. 11 when grown at $40^{\circ}C$ for 32h in the production medium. The protease production profile of the selected mutants was also confirmed by the zymography analysis.