| The development and utilization of fuel ethanol will be an important source of organisms.This topic isusing yeasts from the point of view of alcohol fermentation, produced a plant can effectively use pentoseand yeast cells to hexose fermentation bioethanol.This experiment begin from the distillery yeast,foster screening stochastic spore analysis and appraisalon the haploid get fermentation were performance,screening a fermentation performance relatively betterhaploid A-06.The parents of Saccharomyces cerevisiae A-06and Pachysolen tannophilus B-12protoplast theforming condition of the method,the best training conditions is:concussion training12h of the bacteria,2%helicase and2%cellulase in30℃water bath1h,protoplast formation rate was94.7%and92.5%respectively.The best conditions for cell fusion method the best training conditions is:27℃，pH6.06.5concussion training120rpm.By using two kinds of different inactivation methods for parents of strains protoplast fusion. Usedifferent doses and time ultraviolet inactivated method for parents of yeast alcohol protoplast thoroughlyinactivated,the best training conditions is:30W ultraviolet light from30cm under vertical illuminate1min,the high temperature water bath method to the parents were the yeast B-12protoplast thoroughlyinactivated, best inactivated conditions for65℃water bath1h.Use PEG as melting agent and the parents of the inactivated induction protoplast fusion.After TTC flatcolor reaction at the beginning of the screen, the complex sift and characteristic of the proteinelectrophoresis identification, get a plant to the good performance of xylose fermentation fusion strains ofthe offspring C-05.After determination it Alcohol fermentation degree1.26ml/100ml, parents than B-12improved8.04%.And the ultraviolet ray C-05the mutation, after multiple screening and determination,found that alcohol yield little change, and no success mutagenesis out the ideal strain.And the ultraviolet ray C-05the mutation, after multiple screening and determination, found thatalcohol yield little change, and no success mutagenesis out the ideal strain. Through the different doses of nitrogen ion beam irradiation fusion son, and statistics of the death ratefrom infections after exposure, according to illuminate the processing after death, including dose is8x1015,irradiation time for512s highest death rate, at98.4%.Foster and screening the strains of the offspring, anddetermine the strains of the offspring of the genetic stability.Finally, using the containing different concentration xylose and glucose proportion of medium forbacteria the determination of fermentation performance, and finally a strains were to stabilize the progenyof genetic strain D-08The progeny of the mutants strains after alcohol production rate and increased10%compared to the parents, at the same time, improve the efficiency of the result. |
