scholarly journals Functional expression of horseradish peroxidase in Saccharomyces cerevisiae and Pichia pastoris

2000 ◽  
Vol 13 (5) ◽  
pp. 377-384 ◽  
Author(s):  
Birgit Morawski ◽  
Zhanglin Lin ◽  
Pat Cirino ◽  
Hyun Joo ◽  
Geethani Bandara ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Pichia Pastoris ◽  
Horseradish Peroxidase ◽  
Functional Expression

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.

scholarly journals Functional expression of aryl-alcohol oxidase in Saccharomyces cerevisiae and Pichia pastoris by directed evolution

2018 ◽  
Vol 115 (7) ◽  
pp. 1666-1674 ◽  
Author(s):  
Javier Viña-Gonzalez ◽  
Katarina Elbl ◽  
Xavier Ponte ◽  
Francisco Valero ◽  
Miguel Alcalde
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Pichia Pastoris ◽  
Directed Evolution ◽  
Alcohol Oxidase ◽  
Functional Expression

scholarly journals Functional expression of a heterologous nickel-dependent, ATP-independent urease in Saccharomyces cerevisiae

2015 ◽  
Vol 30 ◽  
pp. 130-140 ◽  
Author(s):  
N. Milne ◽  
M.A.H. Luttik ◽  
H.F. Cueto Rojas ◽  
A. Wahl ◽  
A.J.A. van Maris ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Functional Expression

scholarly journals Purification and characterization of asparaginase II from Saccharomyces cerevisiae cloned in Pichia pastoris: a study on a possible antileukemic drug

2014 ◽  
Vol 8 (S4) ◽  
Author(s):  
Luciana Facchinetti de Castro Girão ◽  
Surza Lucia Gonçalves da Rocha ◽  
Ricardo Sobral Teixeira ◽  
Maria Antonieta Ferrara ◽  
Jonas Perales ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Pichia Pastoris ◽  
Purification And Characterization
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