Coexpression of cellulases in Pichia pastoris as a self-processing protein fusion
메타 데이터
바이오화학분류
바이오플라스틱
플라스틱
바이오정밀화학
용매
화학제품
연료
화장품용 기능성소재
계면활성제⁄증점제
의료용 화학소재
식품첨가제
논문
Coexpression of cellulases in Pichia pastoris as a self-processing protein fusion
학술지
AMB Express
저자명
de Amorim Araú jo, Juliana; Ferreira, Tú lio Cé sar; Rubini, Marciano Ré gis; Duran, Ana Gilhema Gomez; De Marco, Janice Lisboa; de Moraes, Lidia Maria Pepe; Torres, Fernando Araripe Gonç alves
초록
<P>The term cellulase refers to any component of the enzymatic complex produced by some fungi, bacteria and protozoans which act serially or synergistically to catalyze the cleavage of cellulosic materials. Cellulases have been widely used in many industrial applications ranging from food industry to the production of second generation ethanol. In an effort to develop new strategies to minimize the costs of enzyme production we describe the development of a <I>Pichia pastoris</I> strain able to coproduce two different cellulases. For that purpose the <I>eglII</I> (endoglucanase II) and <I>cbhII</I> (cellobiohydrolase II) genes from <I>Trichoderma reesei</I> were fused in-frame separated by the self-processing 2A peptide sequence from the foot-and-mouth disease virus. The protein fusion construct was placed under the control of the strong inducible <I>AOX1</I> promoter. Analysis of culture supernatants from methanol-induced yeast transformants showed that the protein fusion was effectively processed. Enzymatic assay showed that the processed enzymes were fully functional with the same catalytic properties of the individual enzymes produced separately. Furthermore, when combined both enzymes acted synergistically on filter paper to produce cellobiose as the main end-product. Based on these results we propose that <I>P. pastoris</I> should be considered as an alternative platform for the production of cellulases at competitive costs.</P>