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Genetic Transformation of the Yeast Dekkera/Brettanomyces bruxellensis with Non-Homologous DNA
( Miklenic Marina ),( Anamarija Stafa ),( Ana Baji ),( Bojan Zunar ),( Berislav Lisni ),( Ivan Kresimir Svetec ) 한국미생물 · 생명공학회 2013 Journal of microbiology and biotechnology Vol.23 No.5
Yeast Dekkera/Brettanomyces bruxellensis is probably the most common contaminant in wineries and ethanol production processes. The considerable economic losses caused by this yeast, but also its ability to produce and tolerate high ethanol concentrations, make it an attractive subject for research with potential for industrial applications. Unfortunately, efforts to understand the biology of D. bruxellensis and facilitate its broader use in industry are hampered by the lack of adequate procedures for delivery of exogenous DNA into this organism. Here we describe the development of transformation protocols (spheroplast transformation, LiAc/PEG method, and electroporation) and report the first genetic transformation of yeast D. bruxellensis. A linear heterologous DNA fragment carrying the kanMX4 sequence was used for transformation, which allowed transformants to be selected on plates containing geneticin. We found the spheroplast transformation method using 1M sorbitol as osmotic stabilizer to be inappropriate because sorbitol strikingly decreases the plating efficiency of both D. bruxellensis spheroplast and intact cells. However, we managed to modify the LiAc/ PEG transformation method and electroporation to accommodate D. bruxellensis transformation, achieving efficiencies of 0.6-16 and 10-20 transformants/μg DNA, respectively. The stability of the transformants ranged from 93.6% to 100%. All putative transformants were analyzed by Southern blot using the kanMX4 sequence as a hybridization probe, which confirmed that the transforming DNA fragment had integrated into the genome. The results of the molecular analysis were consistent with the expected illegitimate integration of a heterologous transforming fragment.
A new magnetorheological brake`s combined materials design approach
Aleksandar Poznic,Danijela Miloradovic,Anamarija Juhas 대한기계학회 2017 JOURNAL OF MECHANICAL SCIENCE AND TECHNOLOGY Vol.31 No.3
Increase of magnetic flux density intensity, in certain parts of a Magnetorheological (MR) brake, was researched in order to improve MR brake`s overall braking properties. This paper proposes a new combined materials approach. New approach achieved magnetic flux density path routing. To verify the effect, two MR brake prototypes: The conventional one and the combined materials one, have been designed, manufactured and tested. Both prototypes had the same geometry. The construction material’s magnetic properties had to be determined by measurements. To increase the overall braking torque, the combined materials MR brake prototype featured non-magnetic housing materials and has successfully yielded significantly larger overall braking torque compared to the conventional MR brake.
Jasna Canadanovic-Brunet,Gordana Cetkovic,Sonja Djilas,Vesna Tumbas,Gordana Bogdanovi?,Anamarija Mandi?,Sinisa Markov,Dragoljub Cvetkovi?,Vladimir Canadanovi? 한국식품영양과학회 2008 Journal of medicinal food Vol.11 No.1
The aromatic herb Melissa officinalisL. can be used as an easily accessible source of natural antioxidants andas a possible food supplement and as a phytochemical. Radical scavenging, antibacterial, and antiproliferative activities of pe-troleum ether, chloroform, ethyl acetate, n-butanol, and water extracts of M. officinalis L. extracts were investigated. The re-sults of antioxidative activity, obtained by electron spin resonance spectroscopy, confirmed that investigated extracts sup-pressed the formation of 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl, and lipid peroxyl radicals in all investigated systemsin a dose-dependent manner. The maximum DPPH and hydroxyl radical scavenging activities (SADPPH. SAOH . 100%)were achieved in the presence of n-butanol extract at concentrations of 0.4 mg/mL and 0.5 mg/mL, respectively. The highestlipid peroxyl scavenging activity (93.20%) was observed at a higher concentration (5 mg/mL) ofn-butanol extract in the lipidperoxidation system. The most effective antibacterial activities were expressed by petroleum ether and ethyl acetate extractson Sarcina lutea. Chloroform extract showed the strongest antiproliferative effect with 50% inhibitory concentration valuesof 0.09 mg/mL and 0.10 mg/mL for HeLa and MCF-7 cell lines, respectively. The present study demonstrated the high phe-nolic content and radical scavenging, antibacterial, and antiproliferative activities of extracts of M. officinalisL. originatingfrom Serbia.