• 제목/요약/키워드: Starch mutant

검색결과 52건 처리시간 0.032초

Lipase Activity and Tacrolimus Production in Streptomyces clavuligerus CKD 1119 Mutant Strains

  • Kim, Hyung-Soo;Park, Young-In
    • Journal of Microbiology and Biotechnology
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    • 제17권10호
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    • pp.1638-1644
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    • 2007
  • The effect of carbon sources on tacrolimus production by a mutant strain of Streptomyces clavuligerus CKD 1119, an isolate from soil, was examined. Among the carbohydrates and oils tested in this work, a mixed carbon source of soluble starch and com oil was the best. An analysis of the culture kinetics also showed that, in contrast to the carbohydrates, the com oil was consumed later in the antibiotic production phase, implying that the oil substrate was the principal carbon source for the biosynthesis of tacrolimus, and this was directly proven by experiments using $^{14}C$-glucose and $^{14}C$-oleate substrates. Furthermore, com oil induced the formation of lipase by the mutant strain, whereas the addition of glucose significantly repressed lipase activity. The lipase activity exhibited by the FK-506-overproducing mutants was also observed to be directly proportional to their tacrolimus yield, indicating that a high lipase activity is itself a crucial factor for tacrolimus production. A feasibility study with a 200-1 pilot-scale fermentor and the best strain (Tc-XII-15322) identified in this work revealed a high volumetric and specific productivity of about 495 mg/l and 0.34 mg/mg dry mycelium, respectively.

Improved Production, and Purification of Aclacinomycin A from Streptomyces lavendofoliae DKRS

  • Kim, Wan-Seop;Youn, Deok-Joong;Cho, Won-Tae;Kim, Myung-Kuk;Kim, Hak-Ryul;Rhee, Sang-Ki;Choi, Eui-Sung
    • Journal of Microbiology and Biotechnology
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    • 제5권5호
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    • pp.297-301
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    • 1995
  • An anthracycline antibiotic, aclacinomycin A (aclarubicin), was produced from a mutant strain of Streptomyces lavendofoliae. The mutant strain which showed a 4-fold higher productivity of aclacinomycin A compared with the parent strain was also found to produce a significantly higher amount of aclacinomycin A than the reported production strain, Streptomyces galilaeus. The aclacinomycin A was produced up to 125 mg/l using potato starch and soybean meal as carbon and nitrogen sources, respectively, on a 3 liter scale fermentation in a 5 liter jar fermentor. The mutant strain also produced significant amount of aclacinomycins Band Y. Aclacinomycin A was isolated from the culture broth by solvent extractions and further purified by silica gel column chromatography. The yield of aclacinomycin A with over 99$%$ purity was found to be over 60$%$ starting from 3 liters of culture broth.

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Changes in Optimum pH and Thermostability of $\alpha$-amylase from Bacillus licheniformis by Site-directed Mutagenesis of His 235 and Asp 328

  • Kim, Mi-Sook;Lee, Sang-Kyou;Jung, Han-Seung;Yang, Chul-Hak
    • Bulletin of the Korean Chemical Society
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    • 제15권10호
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    • pp.832-835
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    • 1994
  • The ${alpha}$-amylase gene of Bacillus licheniformis has been cloned and two mutant ${alpha}$-amylase genes of which histidine 235 was changed to glutamine (H235Q) and aspartic acid 328 to glutamic acid (D328E) have been produced by site-directed mutagenesis. The kinetic parameters, optimum pH and thermostability of wild type(WT) and these two mutant amylases expressed in E. coli MC1061 have been compared after purification. The $K_m$ values of WT, H235Q and D328E ${alpha}$-amylases were 0.22%, 0.73%, and 0.80% respectively, when using starch as the substrate. The $V_max$ values of wild type ${alpha}$ -amylase and mutant ${alpha}$-amylases were 0.6-0.7%/minute, and did not show any significant differences among them. The optimum pH of D328E ${alpha}$-amylase was shifted to more acidic pH. Also, the thermostability of H235Q ${alpha}$-amylase was increased compared to the wild type ${alpha}$-amylase.

Screening Wheat Germplasm for high Amylose Content

  • Kyeong-Hoon Kim;Jinhee Park;Mina Kim;Kyeong-Min Kim;Yurim Kim;Go Eun Lee;Chon-Sik Kang;Jiyoung Shon;Jong-Min Ko
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2022년도 추계학술대회
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    • pp.300-300
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    • 2022
  • Wheat (Triticum spp.) is an important source of food worldwide and the focus of considerable efforts to identify new combinations of genetic diversity for crop improvement. In particular, wheat starch composition is a major target for changes that could benefit human health. Starches with increased levels of amylose are of interest because of the correlation between high amylose content and elevated levels of resistant starch, which has been shown to have beneficial effects on health for combating obesity and diabetes. In this study, high amylose wheat germplasms from other countries were collected and cultivated in Korea, and then the content of amylose was evaluated, we examined amylose content in 614 wheat germplasm. Furthermore, amylose content was validated using several milling processes such as roller, hammer, and grinding mill. As a result, the amylose content distribution was divided into five groups. The range of the amylose levels in whole wheat flour was 18.3% to 29.6%. In addition, the mutant lines were screened for high amylose, and two mutant lines (WX-1046 and WX-1074) exhibited a comparable amylose content to Keumkang whole wheat (19.6%). It has been established that high amylose indicated SS IIa null and necessitate GBSS. Based on these findings, it may be helpful to develop high amylose wheat germplasm and production techniques, particularly in Korea.

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NTG에 의한 Amylase활성이 높은 누룩사상균의 변이주의 분리 (Isolation of Mutants Overproducing Amylase from Nuruk Fungi by NTG)

  • 정혁준;김영숙;유대식
    • 한국식품영양과학회지
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    • 제29권6호
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    • pp.987-994
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    • 2000
  • 한국 전통누룩으로부터 분리.동정된 Asp. coreanus NR 15-1, Asp. oryzae NR 15-3, 그리고 Asp. oryzae NR 2-5로부터 amylase활성이 높고 세포융합에 사용될 변이주를 분리하고 그의 특성을 검토했다. Aspergillus 속 사상균을 변이처리하기 위한 변이원으로는 NTG를 사용하였으며 Asp. coreanus NR 15-1로부터 총 15종의 변이주중 8종의 영양요구변이주, Asp. oryzae NR 15-3 으로부터 총 5종중 3종의 영양요구변이주를, Asp. oryzae NR 2-5로부터 총 6종중 2종의 영양요구변이주를 분리하였다. 이들 영양요구변이주는 최소배지 에서는 생육하지 않았으며 특정 아미노산과 핵산염기 등을 배지에 첨가했을 경우에만 생육하였다. 분리된 변이주의 당화력, 액화력 및 산 생성능 등을 검토한 결과, 산 생성능이 우수한 균주인 Asp. coreanus NR 15-1로부터 glucoamylase 활성이 높은 \ulcorner(Arg. ̄) 변이주와 Asp. oryzae NR 2-5로 부터 $\alpha$-amylase 활성이 우수한 영양요구변이주인 Z6(Ade. ̄) 변이주를 세포융합에 사용이 가능한 변이주라 사료되었다. 앞으로 이 들의 변이주를 대상으로 세포융합을 통해 우수한 사상균주를 획득할 수 있는 기초적 자료가 됨으로서 전통누룩의 과학화에 기여할 뿔만 아니라 외국 주류와 경쟁력이 있고 과학화된 전통주류의 개발이 가능하리 라 사료된다.

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$\alpha$-Amylase 생성균주 Bacillus sp. AIV 1940의 분리, 특성 및 합성폐수분해능 (Isolation and Characterization of $\alpha$-Amylase Producing Bacillus sp. AIV 1940 and Properties of Starch Synthetic Wastewater Degradation)

  • 박형수;김무훈;양선영;조미영;고범준;박용근
    • 미생물학회지
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    • 제38권1호
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    • pp.1-6
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    • 2002
  • 전분가공폐수처리장 활성슬러지와 논, 밭 토양에서 전분성분을 분해하는 $\alpha$-amylase를 생성하는 균주들을 분리 선별하였다. 이중 효소활성이 우수한 AIV 19를 형태학적 분석 및 API system 과 지방산분석(MIDI) 방법을 이용한 결과 Bacillus 속으로동정되었다. $\alpha$-amylase 생성능을 향상시키기 위해 nitrosoguanidine (NTG)처리를 한 균주 중 선별된 AIV 1940균주의 효소활성능이 원 균주에 비해 1.8배 향상되었다. 분리균주는 그람양성가균으로 길이가 2.3-5 $\mu$ m폭이 0.8-1$\mu$m이었고, Bioscreen C 시스템을 이용한 균주성장능 평가에서는 전분이 3 g/ι농도에서 성장능이 증가되었다. 전분합성폐수를 이용한 유기물제거실험에서는 초기 CODcr가 4,455 mg/ι인 고농도 합성폐수를 1일 3일 반응 후 각각 40.2% 72.3%의 유기물 제거율을 나타내었다.

배유돌연변이체 쌀전분의 이화학적 특성 비교 (Physicochemical Properties of Starch Granules from Endosperm Mutants in Rice)

  • 강미영;한지연;남석현
    • 한국식품과학회지
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    • 제32권2호
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    • pp.258-264
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    • 2000
  • 8품종 변이체 쌀로부터 전분을 분리하여 몇몇 이화학적 특성을 비교하였다. 일반품종 및 저아밀로오스 변이체 쌀 전분입자의 형태는 다변형인데 비해서 shr 및 분질미 전분입자의 형태는 구형을 나타내고 있었다. 남풍 CB243의 호화개시온도 및 호화종료온도가 특히 높았으며, 분질미 및 남풍 EM90의 호화엔탈피가 다른 품종들에 비해서 높았다. 15% $H_2SO_4$에 대한 가수분해도는 shr.이 가장 높았으며, 남풍 CB243이 가장 낮았다. X선 회절도는 8품종 모두 전형적인 A형이었다. 저아밀로오스 유전자가 도입된 품종일수록 glucoamylase에 의한 효소 가수분해도는 증가하고 있었다.

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Improvement of Fungal Cellulase Production by Mutation and Optimization of Solid State Fermentation

  • Vu, Van Hanh;Pham, Tuan Anh;Kim, Keun
    • Mycobiology
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    • 제39권1호
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    • pp.20-25
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    • 2011
  • Spores of Aspergillus sp. SU14 were treated repeatedly and sequentially with $Co^{60}$ ${\gamma}$-rays, ultraviolet irradiation, and N-methyl-N'-nitro-N-nitrosoguanidine. One selected mutant strain, Aspergillus sp. SU14-M15, produced cellulase in a yield 2.2-fold exceeding that of the wild type. Optimal conditions for the production of cellulase by the mutant fungal strain using solid-state fermentation were examined. The medium consisted of wheat-bran supplemented with 1% (w/w) urea or $NH_4Cl$, 1% (w/w) rice starch, 2.5 mM $MgCl_2$, and 0.05% (v/w) Tween 80. Optimal moisture content and initial pH was 50% (v/w) and 3.5, respectively, and optimal aeration area was 3/100 (inoculated wheat bran/container). The medium was inoculated with 25% 48 hr seeding culture and fermented at $35^{\circ}C$ for 3 days. The resulting cellulase yield was 8.5-fold more than that of the wild type strain grown on the basal wheat bran medium.

Expression of Bacillus macerans Cyclodextrin Glucanotransferase in Bacillus subtilis

  • Kim, Chang-Sup;Han, Nam-Soo;Kweon, Dae-Hyuk;Seo, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • 제9권2호
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    • pp.230-233
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    • 1999
  • A plasmid vector was constructed for the expression and secretion of Bacillus macerans cyclodextrin glucanotransferase (CGTase) in Bacillus subtilis. The vector, pUBACGT, was composed of the ribosome-binding sequence, signal sequence, and cgt gene from B. macerans under the control of amyR2, the promoter of amyE gene coding for $\alpha$-amylase from B. subtilis var. natto. Bacillus subtilis LKS88, a mutant strain lacking genes for an amylase and two proteases, was used as a host for the transformation of the plasmid vector. The transformants were selected on kanamycin-containing Luria-Bertani plates. The starch hydrolyzing activity was observed on the starch-containing plates by the iodine method and cyclodextrin-forming activity was detected in the culture medium. A SDS-PAGE analysis showed that most of the expressed CGTase in the recombinant B. subtilis was secreted into the medium at a high expression level.

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보리 화학돌연변이제 처리 $M_2$ 유묘의 Isozyme band pattern 차이에 의한 돌연변이율 검정 (Evaluation of Mutation Rate by Differences of Isozyme Band Patterns on $M_2$ Seedling Treated with Chemical Mutagen in Barley)

  • Bon Cheol, Koo;M., Kucharska
    • 한국작물학회지
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    • 제42권2호
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    • pp.214-219
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    • 1997
  • 세 유형의 화학 돌연변이제 처리(1.5mol Na$N_2$ +0.75mol MNH, 0.75mo1 MNH+0.75mo1 M-NH 및 0.5mol MNH+0.5mol MNH)를 한 M$_2$ 유묘를 이용, Est 1, 2, 4, 5, GOT 1, 2, 3, LAP 1, 2 등의 isozyme band pattern에 대한 돌연변이율을 조사하였다. 1. 세 가지 돌연변이제로 처리된 M$_2$식물체에 대한 돌연변이율 조사를 실시하였는데 엽록체에 대한 돌연변이율은 폴랜드 품종인 Dema에서는 3.3%였고 사천006에서는 1.8%로 나타났다. 2. Esterase유전자좌에 대한 돌연변이율은 Dema에서 3.5%로 사천 006의 0%보다 높았으며 GOT와 LAP에서는 두 품종간 차이가 없었다. 3. 세 가지 isozyme중 대개의 돌연번이체가 Es-terase에 관계된 유전자좌에서 발생하였으며 (75% 차지) 나머지에서의 돌연변이율은 극히 미미하였다.

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