Partial-pKD1 plasmids provide enhanced structural stability for heterologous protein production in Kluyveromyces lactis

1998 ◽  
Vol 49 (4) ◽  
pp. 411-416 ◽  
Author(s):  
H.-P. Hsieh ◽  
N. A. Da Silva
Keyword(s):  
Structural Stability ◽  
Protein Production ◽  
Heterologous Protein ◽  
Kluyveromyces Lactis ◽  
Heterologous Protein Production

scholarly journals Induction by Hypoxia of Heterologous-Protein Production with the KlPDC1 Promoter in Yeasts

2006 ◽  
Vol 73 (3) ◽  
pp. 922-929 ◽  
Author(s):  
Andrea Camattari ◽  
Michele M. Bianchi ◽  
Paola Branduardi ◽  
Danilo Porro ◽  
Luca Brambilla
Keyword(s):  
Protein Production ◽  
Heterologous Protein ◽  
Heterologous Gene Expression ◽  
Kluyveromyces Lactis ◽  
Expression System ◽  
Promoter Sequence ◽  
Heterologous Protein Production ◽  
Heterologous Proteins ◽  
Hypoxic Induction ◽  
Production Ratio

ABSTRACT The control of promoter activity by oxygen availability appears to be an intriguing system for heterologous protein production. In fact, during cell growth in a bioreactor, an oxygen shortage is easily obtained simply by interrupting the air supply. The purpose of our work was to explore the possible use of hypoxic induction of the KlPDC1 promoter to direct heterologous gene expression in yeast. In the present study, an expression system based on the KlPDC1 promoter was developed and characterized. Several heterologous proteins, differing in size, origin, localization, and posttranslational modification, were successfully expressed in Kluyveromyces lactis under the control of the wild type or a modified promoter sequence, with a production ratio between 4 and more than 100. Yields were further optimized by a more accurate control of hypoxic physiological conditions. Production of as high as 180 mg/liter of human interleukin-1β was obtained, representing the highest value obtained with yeasts in a lab-scale bioreactor to date. Moreover, the transferability of our system to related yeasts was assessed. The lacZ gene from Escherichia coli was cloned downstream of the KlPDC1 promoter in order to get β-galactosidase activity in response to induction of the promoter. A centromeric vector harboring this expression cassette was introduced in Saccharomyces cerevisiae and in Zygosaccharomyces bailii, and effects of hypoxic induction were measured and compared to those already observed in K. lactis cells. Interestingly, we found that the induction still worked in Z. bailii; thus, this promotor constitutes a possible inducible system for this new nonconventional host.

scholarly journals Inducible Amplification of Gene Copy Number and Heterologous Protein Production in the Yeast Kluyveromyces lactis

1999 ◽  
Vol 65 (11) ◽  
pp. 4808-4813 ◽  
Author(s):  
Giovanni B. Morlino ◽  
Lorenza Tizzani ◽  
Reinhard Fleer ◽  
Laura Frontali ◽  
Michele M. Bianchi
Keyword(s):  
Copy Number ◽  
Protein Production ◽  
Heterologous Protein ◽  
Gene Dosage ◽  
Kluyveromyces Lactis ◽  
Heterologous Protein Production ◽  
Vector System ◽  
Gene Copy ◽  
Arxula Adeninivorans ◽  
Recombinase Gene

ABSTRACT Heterologous protein production can be doubled by increasing the copy number of the corresponding heterologous gene. We constructed a host-vector system in the yeast Kluyveromyces lactis that was able to induce copy number amplification of pKD1 plasmid-based vectors upon expression of an integrated copy of the plasmid recombinase gene. We increased the production and secretion of two heterologous proteins, glucoamylase from the yeast Arxula adeninivorans and mammalian interleukin-1β, following gene dosage amplification when the heterologous genes were carried by pKD1-based vectors. The choice of the promoters for expression of the integrated recombinase gene and of the episomal heterologous genes are critical for the mitotic stability of the host-vector system.

New Promoters to Improve Heterologous Protein Production in Aspergillus vadensis

2014 ◽  
Vol 3 (3) ◽  
pp. 244-251 ◽  
Author(s):  
Helena Culleton ◽  
Ourdia Bouzid ◽  
Vincent McKie ◽  
Ronald Vries
Keyword(s):  
Protein Production ◽  
Heterologous Protein ◽  
Heterologous Protein Production

Absence of See1p, a widely conservedSaccharomyces cerevisiaeprotein, confers both deficient heterologous protein production and endocytosis

Yeast ◽  
2008 ◽  
Vol 25 (12) ◽  
pp. 871-877 ◽  
Author(s):  
Cristina Martín-Granados ◽  
Sean-Patrick Riechers ◽  
Ulf Stahl ◽  
Christine Lang
Keyword(s):  
Protein Production ◽  
Heterologous Protein ◽  
Heterologous Protein Production

scholarly journals State and specific growth estimation in heterologous protein production by Pichia pastoris

AIChE Journal ◽  
2011 ◽  
Vol 58 (10) ◽  
pp. 2966-2979 ◽  
Author(s):  
José M. Barrigón ◽  
Ramon Ramon ◽  
Isabel Rocha ◽  
Francisco Valero ◽  
Eugénio C. Ferreira ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Protein Production ◽  
Heterologous Protein ◽  
Specific Growth ◽  
Heterologous Protein Production ◽  
Growth Estimation

Heterologous protein production using thePichia pastoris expression system

Yeast ◽  
2005 ◽  
Vol 22 (4) ◽  
pp. 249-270 ◽  
Author(s):  
Sue Macauley-Patrick ◽  
Mariana L. Fazenda ◽  
Brian McNeil ◽  
Linda M. Harvey
Keyword(s):  
Protein Production ◽  
Heterologous Protein ◽  
Expression System ◽  
Heterologous Protein Production ◽  
Pastoris Expression System
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