• Title/Summary/Keyword: systems biotechnology

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Somatic embryogenesis and plant regeneration in zygotic embryo explant cultures of rugosa rose

  • Kim, Suk Weon;Oh, Myung Jin;Liu, Jang R.
    • Plant Biotechnology Reports
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    • v.3 no.3
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    • pp.199-203
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    • 2009
  • Rugosa rose (Rosa rugosa) is cultivated as a garden flower and an important genetic resource for the breeding of roses (R. hybrida). This study describes culture conditions for high frequency plant regeneration from zygotic embryo explants via somatic embryogenesis in rugosa rose. Mature zygotic embryo, cotyledon, and radicle explants formed embryogenic calluses at frequencies of 38, 6.7, and 8.8% when cultured on half-strength Murashige and Skoog medium (${\frac{1}{2}}MS$) supplemented with 2.26, 9.05, and $9.05{\mu}M$ 2,4-dichlorophenoxyacetic acid, respectively. Embryogenic calluses produced numerous somatic embryos, which then developed into plantlets on ${\frac{1}{2}}MS$ without growth regulators. Regenerated plantlets were grown to whole plants in a growth chamber.

Glucose Transport through N-Acetylgalactosamine Phosphotransferase System in Escherichia coli C Strain

  • Kim, Hyun Ju;Jeong, Haeyoung;Lee, Sang Jun
    • Journal of Microbiology and Biotechnology
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    • v.32 no.8
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    • pp.1047-1053
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    • 2022
  • When ptsG, a glucose-specific phosphotransferase system (PTS) component, is deleted in Escherichia coli, growth can be severely poor because of the lack of efficient glucose transport. We discovered a new PTS transport system that could transport glucose through the growth-coupled experimental evolution of ptsG-deficient E. coli C strain under anaerobic conditions. Genome sequencing revealed mutations in agaR, which encodes a repressor of N-acetylgalactosamine (Aga) PTS expression in evolved progeny strains. RT-qPCR analysis showed that the expression of Aga PTS gene increased because of the loss-of-function of agaR. We confirmed the efficient Aga PTS-mediated glucose uptake by genetic complementation and anaerobic fermentation. We discussed the discovery of new glucose transporter in terms of different genetic backgrounds of E. coli strains, and the relationship between the pattern of mixed-acids fermentation and glucose transport rate.

Study of the Rheological Properties of a Fermentation Broth of the Fungus Beauveria bassiana in a Bioreactor Under Different Hydrodynamic Conditions

  • Nunez-Ramirez, Diola Marina;Medina-Torres, Luis;Valencia-Lopez, Jose Javier;Calderas, Fausto;Lopez-Miranda, Javier;Medrano-Roldan, Hiram;Solis-Soto, Aquiles
    • Journal of Microbiology and Biotechnology
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    • v.22 no.11
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    • pp.1494-1500
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    • 2012
  • Fermentation with filamentous fungi in a bioreactor is a complex dynamic process that is affected by flow conditions and the evolution of the rheological properties of the medium. These properties are mainly affected by the biomass concentration and the morphology of the fungus. In this work, the rheological properties of a fermentation with the fungus Beauveria bassiana under different hydrodynamic conditions were studied and the rheological behavior of this broth was simulated through a mixture of carboxymethyl cellulose sodium and cellulose fibers (CMCNa-SF). The bioreactor was a 10 L CSTR tank operated at different stir velocities. Rheological results were similar at 100 and 300 rpm for both systems. However, there was a significant increase in the viscosity accompanied by a change in the consistence index, calculated according to the power law model, for both systems at 800 rpm. The systems exhibited shear-thinning behavior at all stir velocities, which was determined with the power law model. The mixing time was observed to increase as the cellulose content in the system increased and, consequently, the efficiency of mixing diminished. These results are thought to be due to the rheological and morphological similarities of the two fungal systems. These results will help in the optimization of scale-up production of these fungi.

Effects of Automatic Milking Systems on Raw Milk Quality and Milk Fat Properties with or without Feeding Protected Fat (자동착유시스템(AMS) 착유 및 보호지방 첨가 급여가 원유의 품질 및 유지방 특성에 미치는 효과)

  • Moon, Ju Yeon;Chang, Kyeong-Man;Nam, In-Sik;Park, Seong-Min;Oh, Nam Su;Son, Yong-Suk
    • Journal of Dairy Science and Biotechnology
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    • v.32 no.2
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    • pp.63-70
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    • 2014
  • Automatic milking systems (AMS) have been increasingly introduced to Korean dairy farms. However, in comparison with conventional milking systems (CMS), some negative changes in milk quality are being observed. The use of AMS leads to an increase in milking frequency, which in turn might result in higher physical stress on the milk, possibly causing changes in the milk fat globule (MFG) membrane. Therefore, the purpose of this study was to examine the effect of the different milking systems on the milk quality, with a focus on milk fat properties. At the same time, we studied the effect of feeding the dairy cows with protected fat. Raw milk samples were taken monthly from individual cows as well as from bulk tanks at four AMS and four CMS dairy farms. We measured quality-related parameters such as MFG size distribution, free fatty acid content and composition, and acid values. Although most results showed no significant differences with regard to the milking system, we found a relatively high positive correlation between MFG size and milk fat content. Moreover, larger MFG size was observed in the milk when cows had been fed protected fat. The significantly higher (P< 0.05) free fatty acid content of milk observed under this experimental condition could be attributed to higher milking frequency as a result of using AMS.

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In Vitro Evolution of Lipase B from Candida antarctica Using Surface Display in Hansenula polymorpha

  • Kim, So-Young;Sohn, Jung-Hoon;Pyun, Yu-Ryang;Yang, In-Seok;Kim, Kyung-Hyun;Choi, Eui-Sung
    • Journal of Microbiology and Biotechnology
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    • v.17 no.8
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    • pp.1308-1315
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    • 2007
  • Lipase B from Candida antarctica (CalB) displayed on the cell surface of H. polymorpha has been functionally improved for catalytic activity by molecular evolution. CalB was displayed on the cell surface by fusing to a cell-wall anchor motif (CwpF). A library of CalB mutants was constructed by in vivo recombination in H. polymorpha. Several mutants with increased whole-cell CalB activity were acquired from screening seven thousand transformants. The two independent mutants CalB 10 and CalB 14 showed an approximately 5 times greater whole-cell activity than the wild-type. When these mutants were made as a soluble form, CalB 10 showed 6 times greater activity and CalB 14 showed an 11 times greater activity compared with the wild-type. Sequence analyses of mutant CALB genes revealed amino acid substitutions of $Leu^{278}Pro$ in CalB10 and $Leu^{278}Pro/Leu^{219}Gln$ in CalB14. The substituted $Pro^{278}$ in both mutants was located near the proline site of the ${\alpha}$10 helix. This mutation was assumed to induce a conformational change in the ${\alpha}$10 helix and increased the $k_{cat}$ value of mutant CalB approximately 6 times. Site-directed mutagenized CalB, LQ ($Leu^{219}Gln$) was secreted into the culture supernatant at an amount of approximately 3 times more without an increase in the CalB transcript level, compared with the wild-type.

Antifungal Mechanism of Action of Lauryl Betaine Against Skin-Associated Fungus Malassezia restricta

  • Do, Eunsoo;Lee, Hyun Gee;Park, Minji;Cho, Yong-Joon;Kim, Dong Hyeun;Park, Se-Ho;Eun, Daekyung;Park, Taehun;An, Susun;Jung, Won Hee
    • Mycobiology
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    • v.47 no.2
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    • pp.242-249
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    • 2019
  • Betaine derivatives are considered major ingredients of shampoos and are commonly used as antistatic and viscosity-increasing agents. Several studies have also suggested that betaine derivatives can be used as antimicrobial agents. However, the antifungal activity and mechanism of action of betaine derivatives have not yet been fully understood. In this study, we investigated the antifungal activity of six betaine derivatives against Malassezia restricta, which is the most frequently isolated fungus from the human skin and is implicated in the development of dandruff. We found that, among the six betaine derivatives, lauryl betaine showed the most potent antifungal activity. The mechanism of action of lauryl betaine was studied mainly using another phylogenetically close model fungal organism, Cryptococcus neoformans, because of a lack of available genetic manipulation and functional genomics tools for M. restricta. Our genome-wide reverse genetic screening method using the C. neoformans gene deletion mutant library showed that the mutants with mutations in genes for cell membrane synthesis and integrity, particularly ergosterol synthesis, are highly sensitive to lauryl betaine. Furthermore, transcriptome changes in both C. neoformans and M. restricta cells grown in the presence of lauryl betaine were analyzed and the results indicated that the compound mainly affected cell membrane synthesis, particularly ergosterol synthesis. Overall, our data demonstrated that lauryl betaine influences ergosterol synthesis in C. neoformans and that the compound exerts a similar mechanism of action on M. restricta.