• 제목/요약/키워드: Culture Conditions

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Hfq and ArcA Are Involved in the Stationary Phase-Dependent Activation of Salmonella Pathogenicity Island 1 (SPI1) Under Shaking Culture Conditions

  • Lim, Sangyong;Yoon, Hyunjin;Kim, Minjeong;Han, Ahreum;Choi, Jihae;Choi, Jeongjoon;Ryu, Sangryeol
    • Journal of Microbiology and Biotechnology
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    • 제23권12호
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    • pp.1664-1672
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    • 2013
  • In Salmonella enterica serovar Typhimurium, many genes encoded within Salmonella pathogenicity island 1 (SPI1) are required to induce intestinal/diarrheal disease. In this study, we compared the expression of four SPI1 genes (hilA, invF, prgH, and sipC) under shaking and standing culture conditions and found that the expression of these genes was highest during the transition from the exponential to stationary phase under shaking conditions. To identify regulators associated with the stationary phase-dependent activation of SPI1, the effects of selected regulatory genes, including relA/spoT (ppGpp), luxS, ihfB, hfq, and arcA, on the expression of hilA and invF were compared under shaking conditions. Mutations in the hfq and arcA genes caused a reduction in hilA and invF expression (more than 2-fold) in the early stationary phase only, whereas the lack of ppGpp and IHF decreased hilA and invF gene expression during the entire stationary phase. We also found that hfq and arcA mutations caused a reduction of hilD expression upon entry into the stationary phase under shaking culture conditions. Taken together, these results suggest that Hfq and ArcA regulate the hilD promoter, causing an accumulation of HilD, which can trigger a stationary phase-dependent activation of SPI1 genes under shaking culture conditions.

Optimal Manufacturing Conditions for Korean Soybean Paste and Soy Sauce, Using Aspergillus oryzae AJ 100 as a Flavor Improver

  • Park, Hea-Kyeung;Kim, Jong-Kyu
    • Food Science and Biotechnology
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    • 제17권1호
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    • pp.208-211
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    • 2008
  • Previously, it has been reported that Aspergillus oryzae can efficiently degrade unpleasant odor components such as butyric acid and 3-methyl butanoic acid from meju, a major ingredient in both Korean soybean paste (doenjang) and soy sauce. In this study, the optimal manufacturing conditions for the production of superior quality Korean soybean paste and soy sauce were determined. Specifically, A. oryzae AJ 100 was utilized to improve the flavor of these products. Mixtures of Korean soybean paste and A. oryzae AJ 100 culture (2 : 1), and of Korean soy sauce and A. oryzae AJ 100 culture (5 : 1), were incubated for 2 weeks at $30^{\circ}C$, and showed improved flavor. Butyric acid and 3-methyl butanoic acid were clearly degraded under these culture conditions.

Astaxanthin 생산을 위한 Phaffia rhodozyma의 변이균주 선발과 최적 배양조건 결정 (Selection of mutant Phaffia rhodozyma and Determination of Optimum Culture Conditions for Astaxanthin Production)

  • 유성선;유연우
    • 한국미생물·생명공학회지
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    • 제29권2호
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    • pp.96-103
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    • 2001
  • Phaffia rhodozyma is the most promising microbial source of astaxanthin production, though wild-type strains are needed to increase the astaxanthin content for commercial production. To increase astaxanthin content for commercial production, a mutant strain of P. rhodozyma was selected and culture conditions of the mutant selected were optimized. P. rhodozyma was treated with mutagenic agent such as NTG, acriflavine, and UV in serial order and carotenoids hyper-producing mutant strain was selected based on the capabilities of cell growth on the agar plate containing chemical inhibitors and carotenoids production. Among the mutants tested, a mutant WS-2 was finally selected. Mutant WS-2 produced 1.26mg carotenoids/g-dry cell weight and this value was about- 4-folds higher than that of wild-type. The optimum culture conditions were $24^{\circ}C$ of temperature, 1.5vvm of aeration and 300rpm of agitation. In the optimized condition, cell and carotenoids concentrations were 7.62g/l and 14.9mg/l, respectively.

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Study on Gloeostereum Inoarnatum 5. Itoetimai - Fermentation Cultivation(Liquid Fermentation)

  • Jie, Tai-Long
    • Plant Resources
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    • 제4권3호
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    • pp.200-205
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    • 2001
  • It was reported in our Previous paper that the fermented products from Gloeostereum incarnatum strongly inhibit the growth of six kinds of bacteria in human bodies. In this paper the appropriated conditions of immersing culture for the strain 8 903 of Gloeostereum incarnatum was analysed. And the output of the hypha and fermentative product was determined or compared. The prelimenaryresults showed that the appropriated conditions for the growth of Gloeostereum incarnatum are: (1)culture medium:glucose 3%; protein peoptne 0.2%; soybeancake power 1% yeast power 0.3%; KH2PO40.05%; MgSO4 0.03%; CaCO3 0.01%; vitamin Bl 0.001%; befor sterilization pH Value of six should be maintained; (2) temperature; 27f ~28f ; (3) time; about 200 hours; (4) ventilation; (30%∼50%)/min. The sigh of the end culture are: pH coming down about 4: remnant glucoses less 1%; amino nitrogens about 20%; time about eight days. In the aforementioned conditions, the output of fermentative product achieve to 2.5∼3g/L.

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STUDY ON GLOEOSTEREUM INOARNATUM S. ITOET IMAI-FERMENTATION CULTIVATION(LIQUID FERMENTATION)

  • Jie, Tai-Long
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2001년도 The 8th International Symposium
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    • pp.74-82
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    • 2001
  • It was reported in our Previous paper that the fermented products from Gloeostereum incarnatum strongly inhibit the growth of six kinds of bacteria in human bodies. In this paper the appropriated conditions of immersing culture for the strain 8 903 of Gloeostereum incarnatum was analysed. And the output of the hypha and fermentative product was determined or compared, The prelimenaryresults showed that the appropriated conditions for the growth of Gloeostereum incarnatum are: (1)culture medium:glucose 3%; protein peoptne 0.2%; soybeancake power 1%, yeast power 0.3%; KH2PO40.05%; MgSO4 0.03%; CaCO3 0.01%; vitamin Bl 0.001%; befor sterilization pH Value of six should be maintained; (2) temperature; 27$^{\circ}C$~28$^{\circ}C$; (3) time; about 200 hours; (4) ventilation; (30%~50%)/min. The sigh of the end culture we: pH coming down about 4: remnant glucoses less 1%, amino nitrogens about 20;, time about eight days. In the aforementioned conditions, the output of fermentative product achieve to 2.5 ~3g/L.

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Optimization of Solid-State Fermentation for Improved Conidia Production of Beauveria bassiana as a Mycoinsecticide

  • Pham, Tuan Anh;Kim, Jeong-Jun;Kim, Keun
    • Mycobiology
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    • 제38권2호
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    • pp.137-143
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    • 2010
  • The production of conidia of entomopathogenic Beauveria bassiana by solid-state fermentation was studied for the development of a biocontrol agent against aphid Myzus persicae. The optimal conditions for conidia production on polished white rice were 40% moisture content, $25^{\circ}C$ culture temperature, 2-day-old seeding culture grown in 3% corn meal, 2% rice bran, 2% corn steep powder medium, initial conidia concentration of $10^7$ conidia/g in the wet rice, 10% inoculum size, and use of a polyethylene bag as a container. The polyethylene bag containing inoculated rice was hand-shaken every 12 hr during fermentation. Using optimal conditions, the maximum conidia production obtained was 4.05 g conidia/100 g dry rice after 14 days of cultivation, a rate 2.83 times higher than conidia yield of pre-optimization.

Optimization of Culture Conditions and Bench-Scale Production of $_L$-Asparaginase by Submerged Fermentation of Aspergillus terreus MTCC 1782

  • Gurunathan, Baskar;Sahadevan, Renganathan
    • Journal of Microbiology and Biotechnology
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    • 제22권7호
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    • pp.923-929
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    • 2012
  • Optimization of culture conditions for L-asparaginase production by submerged fermentation of Aspergillus terreus MTCC 1782 was studied using a 3-level central composite design of response surface methodology and artificial neural network linked genetic algorithm. The artificial neural network linked genetic algorithm was found to be more efficient than response surface methodology. The experimental $_L$-asparaginase activity of 43.29 IU/ml was obtained at the optimum culture conditions of temperature $35^{\circ}C$, initial pH 6.3, inoculum size 1% (v/v), agitation rate 140 rpm, and incubation time 58.5 h of the artificial neural network linked genetic algorithm, which was close to the predicted activity of 44.38 IU/ml. Characteristics of $_L$-asparaginase production by A. terreus MTCC 1782 were studied in a 3 L bench-scale bioreactor.

랫드 배아 조작 효율 향상을 위한 배양 조건 (Culture Conditions for Improving Manipulation Efficiency of Rat Embryo)

  • 이지민
    • 한국환경과학회지
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    • 제32권3호
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    • pp.173-179
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    • 2023
  • Rats are one of the most widely used animals in biomedical sciences because their metabolism and physiology are comparable to humans. In recent years, gene-targeted models have been developed using various animal species utilizing engineered nucleases such as clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated gene (Cas). It has recently become possible to efficiently transfect CRISPR/Cas into embryos via electroporation. However, electroporation can damage fertilized eggs; therefore, it is important to determine the optimal embryo culture conditions. A standardized approach for routine and reproducible rat transgenesis will render rat models more accessible for research. We performed experiments to obtain rat embryos with efficient superovulation and synchronization, and to investigate the appropriate medium conditions for pronuclear stage embryos subjected to electroporation stimulation for the introduction of engineered nuclease.

Rhamnolipid Production in Batch and Fed-batch Fermentation Using Pseudomonas aeruginosa BYK-2 KCTC 18012P

  • Lee, Kyung-Mi;Hwang, Sun-Hee;Ha, Soon-Duck;Jang, Jae-Hyuk;Lim, Dong-Jung;Kong, Jai-Yul
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제9권4호
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    • pp.267-273
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    • 2004
  • The optimization of culture conditions for the bacterium Pseudomonas aeruginosa BYK-2 KCTC 18012P, was performed to increase its rhamnolipid production. The optimum level for carbon, nitrogen sources, temperature and pH, for rhamnolipid production in a flask, were identified as 25 g/L fish oil, 0.01% (w/v) urea, 25 and pH 7.0, respectively. Optimum conditions for batch culture, using a 7-L jar fermentor, were 200 rpm of agitation speed and a 2.0 L/min aeration rate. Under the optimum conditions, on fish oil for 216 h, the final cell and rhamnolipid concentrations were 5.3 g/L and 17.0 g/L respectively. Fed-batch fermentation, with different feeding conditions, was carried out in order to increase, cell growth and rhamnolipid production by the Pseudomonas aeruginosa, BYK-2 KCTC 18012P. When 2.5 g of fish oil and 100 mL basal salts medium, containing 0.01 % (w/v) urea, were fed intermittently during the fermentation, the final cell and rhamnolipid concentrations at 264 h, were 6.1 and 22.7 g/L respectively. The fed-batch culture resulted in a 1.2-fold increase in the dry cell mass and a 1.3-fold increase in rhamnolipid production, compared to the production of the batch culture. The rhamnolipid production-substrate conversion factor (0.75 g/g) was higher than that of the batch culture (0.68 g/g).

Insertional mutations exhibiting high cell-culture density HCD phenotypes are enriched through continuous subcultures in Chlamydomonas reinhardtii

  • Thung, Leena;He, Jing;Zhu, Qingling;Xu, Zhenyu;Liu, Jianhua;Chow, Yvonne
    • ALGAE
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    • 제33권1호
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    • pp.127-141
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    • 2018
  • Low efficiency in microalgal biomass production was largely attributed to the low density of algal cell cultures. Though mutations that reduced the level of chlorophyll or pigment content increased efficiency of photon usage and thus the cell-culture density under high-illumination growth conditions (e.g., >$500{\mu}mol\;photon\;m^{-2}\;s^{-1}$), it was unclear whether algae could increase cell-culture density under low-illumination conditions (e.g., ${\sim}50{\mu}mol\;photon\;m^{-2}\;s^{-1}$). To address this question, we performed forward genetic screening in Chlamydomonas reinhardtii. A pool of >1,000 insertional mutants was constructed and subjected to continuous subcultures in shaking flasks under low-illumination conditions. Complexity of restriction fragment length polymorphism (RFLP) pattern in cultures indicated the degree of heterogeneity of mutant populations. We showed that the levels of RFLP complexity decreased when cycles of subculture increased, suggesting that cultures were gradually populated by high cell-culture density (HCD) strains. Analysis of the 3 isolated HCD mutants after 30 cycles of subcultures confirmed that their maximal biomass production was 50-100% higher than that of wild type under low-illumination. Furthermore, levels of chlorophyll content in HCD mutant strains were similar to that of wild type. Inverse polymerase chain reaction analysis identified the locus of insertion in two of three HCD strains. Molecular and transcriptomic analyses suggested that two HCD mutants were a result of the gain-of-function phenotype, both linking to the abnormality of mitochondrial functions. Taken together, our results demonstrate that HCD strains can be obtained through continuous subcultures under low illumination conditions.