Optimization of the Functional Expression of Coprinus cinereus Peroxidase in Pichia pastoris by Varying the Host and Promoter

2009 ◽  
Vol 19 (9) ◽  
pp. 966-971 ◽  
Author(s):  
Su-Jin Kim
Keyword(s):  
Pichia Pastoris ◽  
Functional Expression ◽  
Coprinus Cinereus ◽  
Coprinus Cinereus Peroxidase

Functional expression of Coprinus cinereus peroxidase in Pichia pastoris

2009 ◽  
Vol 44 (7) ◽  
pp. 731-735 ◽  
Author(s):  
Su Jin Kim ◽  
Jeong Ah Lee ◽  
Keehoon Won ◽  
Yong Hwan Kim ◽  
Bong Keun Song
Keyword(s):  
Pichia Pastoris ◽  
Functional Expression ◽  
Coprinus Cinereus ◽  
Coprinus Cinereus Peroxidase

Functional expression of porcine interferon-α using a combinational strategy in Pichia pastoris GS115

2019 ◽  
Vol 122 ◽  
pp. 55-63 ◽  
Author(s):  
Huahua He ◽  
Chao Zhai ◽  
Meng Mei ◽  
Yi Rao ◽  
Yao Liu ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Functional Expression ◽  
Interferon Α ◽  
Pastoris Gs115 ◽  
Pichia Pastoris Gs115

scholarly journals Optimization of an Activity Assay of Coprinus Cinereus Peroxidase

2018 ◽  
Vol 61 (0) ◽  
Author(s):  
Bing Xue Dong ◽  
Yu Hong Zhang ◽  
Gang Li ◽  
Jin Tao Fang ◽  
Ji Huan Zheng ◽  
...  
Keyword(s):  
Coprinus Cinereus ◽  
Activity Assay ◽  
Coprinus Cinereus Peroxidase

Cloning, functional expression and characterization of Aspergillus sulphureus β-mannanase in Pichia pastoris

2007 ◽  
Vol 128 (3) ◽  
pp. 452-461 ◽  
Author(s):  
Xiaoling Chen ◽  
Yunhe Cao ◽  
Yuhua Ding ◽  
Wenqing Lu ◽  
Defa Li
Keyword(s):  
Pichia Pastoris ◽  
Functional Expression ◽  
Aspergillus Sulphureus

scholarly journals Functional expression of the lactate permease Jen1p of Saccharomyces cerevisiae in Pichia pastoris

2003 ◽  
Vol 376 (3) ◽  
pp. 781-787 ◽  
Author(s):  
Isabel SOARES-SILVA ◽  
Dorit SCHULLER ◽  
Raquel P. ANDRADE ◽  
Fátima BALTAZAR ◽  
Fernanda CÁSSIO ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Acetic Acid ◽  
Lactic Acid ◽  
Pichia Pastoris ◽  
Plasma Membranes ◽  
Membrane Vesicle ◽  
Dry Weight ◽  
Functional Expression ◽  
Acid Uptake ◽  
Methanol Induction

In Saccharomyces cerevisiae the activity for the lactate–proton symporter is dependent on JEN1 gene expression. Pichia pastoris was transformed with an integrative plasmid containing the JEN1 gene. After 24 h of methanol induction, Northern and Western blotting analyses indicated the expression of JEN1 in the transformants. Lactate permease activity was obtained in P. pastoris cells with a Vmax of 2.1 nmol·s−1·mg of dry weight−1. Reconstitution of the lactate permease activity was achieved by fusing plasma membranes of P. pastoris methanol-induced cells with Escherichia coli liposomes containing cytochrome c oxidase, as proton-motive force. These assays in reconstituted heterologous P. pastoris membrane vesicles demonstrate that S. cerevisiae Jen1p is a functional lactate transporter. Moreover, a S. cerevisiae strain deleted in the JEN1 gene was transformed with a centromeric plasmid containing JEN1 under the control of the glyceraldehyde-3-phosphate dehydrogenase constitutive promotor. Constitutive JEN1 expression and lactic acid uptake were observed in cells grown on either glucose and/or acetic acid. The highest Vmax (0.84 nmol·s−1·mg of dry weight−1) was obtained in acetic acid-grown cells. Thus overexpression of the S. cerevisiae JEN1 gene in both S. cerevisiae and P. pastoris cells resulted in increased activity of lactate transport when compared with the data previously reported in lactic acid-grown cells of native S. cerevisiae strains. Jen1p is the only S. cerevisiae secondary porter characterized so far by heterologous expression in P. pastoris at both the cell and the membrane-vesicle levels.

Sign in / Sign up

Export Citation Format

Share Document