Protoplast fusion is a common approach used to improve fermentation of industrial yeast strains. Molecular study using DNA content, RAPD, DNA sequences and protein profile were applied to differentiate between S. cerevisiae and P. stipitis ATCC 58785 and their fusants from protoplast fusion for improvement of biofuel production from biomass. Fusants were showed higher DNA content than that in the parental strain and DNA concentration obtained for recently generated fusant was generally lower compared to the values expected by the theoretical addition of DNA concentration from the respective parental strains. Fusants were showed new combination of DNA fragment patterns. The DNA sequencing, which includes two non-coding regions designated as the internal transcribed spacers (ITS1 and ITS2) and the 5.8S gene were performed for further confirmation of the fusant nature of fusants. According to genetic similarity and intra-species differentiation, two parent strains and fusants were grouped into two different clusters. S. cerevisiae which correspond to 88% sequence similarity whereas approximately 97% similarity was observed with P. stipitis ATCC 58785.The sequence of the ITS1, ITS2 and the 5.8S gene of each fusants was submitted to Genbank with the NCBI ACESSION NO. In general, the obtained new combination of DNA fragment patterns and the presence of new DNA fragment or the absence of existing parental DNA fragments in the fusant strains compared to their parents could be considered as indicator of nuclear fusion of the two parental nuclei in the fused protoplasts, also differences in polypeptide profile on SDS-PAGE analysis were investigated. The polypeptide profile was seen with reference to a protein ladder (GangNam-STAIN™ Prestained). SDS-PAGE protein analysis of the selected fusants and their parental strains confirmed that all fusant strains acquired and expressed many specific protein bands from the parental strains.
Bioethanol is gaining increasing attention as a clean and renewable fuel because of its major environmental benefits. Efficient bioethanol fermentation requires the selection of superior strains that are capable of ethanol stress tolerance. Yeast can produce ethanol, thereby reflecting its intrinsic ability to tolerate ethanol. This study focuses on ethanol tolerance enhancement of Saccharomyces cerevisiae for ethanol production improvement through protoplast fusion. S. cerevisiae and P. stipitis fusants (ATCC 58785), which can ferment xylose, were isolated. The ability of these isolates tolerate ethanol was investigated by allowing the strains to grow in different ethanol concentrations. Results showed the ability of the fusantsto have an average tolerance to ethanol when compared with the parent strains and fermented glucose in the presence of 6% ethanol. By contrast, the parent strains S. cerevisiae and P. stipitis showed ethanol tolerances of 8 and 4%, respectively. Fusant formation was confirmed by the increased DNA content. This outcome suggests that multiple fusions had occurred and the genetic stability of fusants indicates that F24 and F18 are genetically stable and suitable for industrial production.
Honey is one of the oldest natural medicines known with a very high therapeutic value. Nowadays, in the medical field, several important therapeutic effects of honey have been elucidated. This study was conducted to reveal the antimicrobial activity of the commercially available local Malaysian Trigona sp. honey towards different pathogenic bacteria specifically Staphylococcus aureus, Streptococcus pyogenes, Escherichia coli, and Salmonella typhimurium. The Minimum inhibitory concentration and Minimum bactericidal concentration were determined by the disc-diffusion test and agar well diffusion test. Different concentration of the honey was tested in the disc-diffusion and agar well diffusion test. The results of these tests were in terms of Inhibition zone diameter. The results obtained from the current study are the dilution of different concentration of honey from Trigona sp. are very significant because the only net concentration of both of honey Trigon a sp. possessed antimicrobial properties in term of Minimum inhibitory concentration) and Minimum bactericidal concentration. The result also can say that Trigona sp. honey possessed antibacterial properties and can be used as alternative medicine in the veterinary field in the future.
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