Abstract: In this study,four different heterozygous fibrin scaffold proteins were successfully constructed. Cellulase and its extracellular assembly into a different spatial structure of chimeric designer cellulosome. The thermal stability and enzymatic hydrolysis kinetics of chimeric designer cellulosome were analyzed. The influence of different structures on the hydrolysis activity of designer cellosome and its optimal scaffold proteins structure was explored. Based on the Saccharomyces cerevisiae eby100 surface display system,the optimized scaffold protein was anchored and the performance of simultaneous saccharification and ethanol fermentation were assayed on PSAC. The results show that the enzyme activity of the designer cellosome remains relatively stable at 50 ℃ for 120 hours,the V_max=0.147 mg/（mL·min）,K_m=6.085 mg/mL and the reducing sugar production of ScafI-Ⅳ designer cellosome are all higher than those of the other cellosomes structural,which indicates the ScafI-Ⅳ type has a excellence substrate affinity and hydrolysis property. The maximum ethanol production is 1.12 g/L after 96 h,and the output is 0.263 g/g,which is equivalent to 51.50% of the theoretical value of ethanol yield. This result proves that the enzymolysis performance of the designer cellosome can be improved by structure optimizing of the scaffold proteins,which has reference significance for the research of artificially designing the cellosome.