Scale-Up of a High Cell Density Continuous Culture with Pichia pastoris X-33 for the Constitutive Expression of rh-Chitinase

2001 ◽  
Vol 17 (4) ◽  
pp. 629-633 ◽  
Author(s):  
B.M. Schilling ◽  
J.C. Goodrick ◽  
N.C. Wan
Keyword(s):  
Pichia Pastoris ◽  
Continuous Culture ◽  
Cell Density ◽  
High Cell Density ◽  
Scale Up ◽  
Constitutive Expression ◽  
High Cell

scholarly journals Pichia pastoris: SEL RAGI UNTUK PRODUKSI PROTEIN REKOMBINAN

2018 ◽  
Vol 17 (2) ◽  
Author(s):  
Neng Herawati ◽  
Arizah Kusumawati ◽  
Adi Santoso
Keyword(s):  
Pichia Pastoris ◽  
Recombinant Protein ◽  
Cell Density ◽  
Mammalian Cells ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
High Cell Density ◽  
Expression System ◽  
Scale Up ◽  
High Cell

Pichia pastoris is a group of methylotropic yeast known as a host of expression and protein production which is widely used for biopharmaceutical-based drug production. This yeast can grow fast with a high cell density. Its genetic stability, high cell density, and stress resistance make the development process and scale-up of P. pastoris can increase to a scale of 200,000 liters of culture. In contrast to the expensive and complex development of recombinant protein production in mammalian cells, the development of production in P. pastoris is relatively simple and cheaper. The advantage of P. pastoris as an expression system is that it is able to use methanol as a carbon source by inducing the expression of alcohol oxidase oxidase (AOX) enzyme. Promoter used by this enzyme is also used as a strong promoter for the expression of proteins that we want. Unlike in bacterial and mammalian systems, recombinant protein production in Pichia cells is not contaminated with endotoxins or viruses so it is safer and simplifies the downstream processes in bioproduction. The level of endogenous protein in the low supernatant allows Pichia to cultivate with a high volumetric productivity, therefore the process of protein production becomes very economical. This review provides an overview of several things that must be considered in utilizing P. pastoris as an expression system including the selection of vectors, strains, vector integration mechanisms into the genome, glycosylation processes, and applications in industry.

scholarly journals Poly(3-Hydroxybutyrate) Production from Glycerol by Zobellella denitrificans MW1 via High-Cell-Density Fed-Batch Fermentation and Simplified Solvent Extraction

2009 ◽  
Vol 75 (19) ◽  
pp. 6222-6231 ◽  
Author(s):  
Mohammad H. A. Ibrahim ◽  
Alexander Steinbüchel
Keyword(s):  
Solvent Extraction ◽  
Cell Density ◽  
High Cell Density ◽  
Scale Up ◽  
Downstream Processing ◽  
Production Costs ◽  
Yield Coefficient ◽  
High Cell ◽  
Fed Batch ◽  
High Specific Growth Rate

ABSTRACT Industrial production of biodegradable polyesters such as polyhydroxyalkanoates is hampered by high production costs, among which the costs for substrates and for downstream processing represent the main obstacles. Inexpensive fermentable raw materials such as crude glycerol, an abundant by-product of the biodiesel industry, have emerged to be promising carbon sources for industrial fermentations. In this study, Zobellella denitrificans MW1, a recently isolated bacterium, was used for the production of poly(3-hydroxybutyrate) (PHB) from glycerol as the sole carbon source. Pilot-scale fermentations (42-liter scale) were conducted to scale up the high PHB accumulation capability of this strain. By fed-batch cultivation, at first a relatively high cell density (29.9 ± 1.3 g/liter) was obtained during only a short fermentation period (24 h). However, the PHB content was relatively low (31.0% ± 4.2% [wt/wt]). Afterwards, much higher concentrations of PHB (up to 54.3 ± 7.9 g/liter) and higher cell densities (up to 81.2 ± 2.5 g/liter) were obtained by further fed-batch optimization in the presence of 20 g/liter NaCl, with optimized feeding of glycerol and ammonia to support both cell growth and polymer accumulation over a period of 50 h. A high specific growth rate (0.422/h) and a short doubling time (1.64 h) were attained. The maximum PHB content obtained was 66.9% ± 7.6% of cell dry weight, and the maximum polymer productivity and substrate yield coefficient were 1.09 ± 0.16 g/liter/h and 0.25 ± 0.04 g PHB/g glycerol, respectively. Furthermore, a simple organic solvent extraction process was employed for PHB recovery during downstream processing: self-flotation of cell debris after extraction of PHB with chloroform allowed a convenient separation of a clear PHB-solvent solution from the cells. Maximum PHB recovery (85.0% ± 0.10% [wt/wt]) was reached after 72 h of extraction with chloroform at 30°C, with a polymer purity of 98.3% ± 1.3%.

Cloning, expression, characterization, and high cell-density production of recombinant endo-1,4-β-xylanase from Aspergillus niger in Pichia pastoris

2007 ◽  
Vol 41 (1-2) ◽  
pp. 19-25 ◽  
Author(s):  
Vasimon Ruanglek ◽  
Rutchadaporn Sriprang ◽  
Nakul Ratanaphan ◽  
Pacawadee Tirawongsaroj ◽  
Duriya Chantasigh ◽  
...  
Keyword(s):  
Aspergillus Niger ◽  
Pichia Pastoris ◽  
Cell Density ◽  
High Cell Density ◽  
High Cell ◽  
Expression Characterization
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