Genetic Engineering of Saccharomyces Cerevisiae Using A Novel Approach Global Transcription Machinery Engineering for Enhancing Bioethanol Production
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- gTME, Saccharomyces Cerevisiae, SPT15, Ethanol Tolerance, Error-Prone PCR, Ethanol Production
- El-Rotail, Ashraf; Shia, Gui Yang; Rashed, Marwan M. A.; Al-Farga, Ammar; HuaLi, ; Junhua, Wang; Sun, Yang
- The purpose of our study was investigate from effect new mutations of Saccharomyces cerevisiae which possessing the ability to produce and enhance ethanol yield and characterized by ethanol tolerance which had been obtained by using the new protocol has been developed in our previous study. By molecular biology methods, such as Error-prone PCR, global transcription machinery engineering, DNA hybridization, etc. were used to construct an initial transcription factor and screen the target specific phenotype for obtaining mutations SPT15-Mu genes from the Saccharomyces cerevisiae R-control strain; which have an ability to produce and enhance ethanol yield and characterized by ethanol tolerance. The results showed more than 80 mutant genes were obtained with SPT15 gene that diversely expressed when utilized MnCl2. Mutants recorded the highest ethanol production by using 3 % MnCl2 with improving the yield rate to 60.24 % compared to R-control strain. In conclusions, there are many vehicles that produced by metabolism resulting by the fermentation process; which are considered work as stress factors for yeast, leading to weaken their activities, and eventually cause cell loss. In our study, we managed to get random mutagenesis-Super; which were characterized by a high mutation rate, and simultaneously be stable in front of a lot of large compounds of metabolites produced by the fermentation process. Even be able to grow better than wild-type strain.
Full text: IJRAS_548_FINAL.pdf