Production of minicellulosomes from Clostridium cellulovorans for the fermentation of cellulosic ethanol using engineered recombinant Saccharomyces cerevisiae.
Saccharomyces cerevisiae was engineered for assembly of minicellulosomes by heterologous expression of a recombinant scaffolding protein from Clostridium cellulovorans and a chimeric endoglucanase E from Clostridium thermocellum. The chimeric endoglucanase E fused with the dockerin domain of endoglucanase B from C. cellulovorans was assembled with the recombinant scaffolding protein. The resulting strain was able to ferment amorphous cellulose [carboxymethyl-cellulose (CMC)] into ethanol with the aid of beta-glucosidase 1 produced from Saccharomycopsis fibuligera. The minicellulosomes assembled in vivo retained the synergistic effect for cellulose hydrolysis. The minicellulosomes containing the cellulose-binding domain were purified by crystalline cellulose affinity in a single step. In the fermentation test at 10 g L(-1) initial CMC, approximately 3.45 g L(-1) ethanol was produced after 16 h. The yield (in grams of ethanol produced per substrate) was 0.34 g g(-1) from CMC. This result indicates that a one-step processing of cellulosic biomass in a consolidated bioprocessing configuration is technically feasible by recombinant yeast cells expressing functional minicellulosomes.
Hyeon
Jeong-eun
JE
School of Life Science and Biotechnology, Korea University, Seoul, Korea.
Yu
Kyung-Ok
KO
Suh
Dong Jin
DJ
Suh
Young-Woong
YW
Lee
Sung Eun
SE
Lee
Jinwon
J
Han
Sung Ok
SO
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002482:Cellulose / Q000378:metabolism*; D044603:Cellulosomes / Q000235:genetics / Q000378:metabolism*; D046974:Clostridium cellulovorans / Q000201:enzymology* / Q000235:genetics; D048013:Acetivibrio thermocellus / Q000201:enzymology* / Q000235:genetics; D000431:Ethanol / Q000378:metabolism*; D005285:Fermentation; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D000431:Ethanol; D002482:Cellulose