Industrial production of heterologous proteins by fed-batch cultures of the yeast Saccharomyces cereuisiae

Mendoza-Vega, O.

doi:10.1016/0168-6445(94)90070-1

Cited by 27 publications

(34 citation statements)

References 113 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Fed-batch fermentation is a useful tool for achieving high productivity in the industrial-scale production of foreign proteins in microorganism or mammalian cell hosts using a stirred fermentor (Bibila and Robinson, 1995;Minihane and Brown, 1986;Mondoza-Vega et al, 1994). One of the main reasons is that it allows the substrate concentration in the culture broth to be adjusted to a value suitable for cell growth and production.…”

Section: Introductionmentioning

confidence: 99%

Optimization of human serum albumin production in methylotrophic yeast Pichia pastoris by repeated fed‐batch fermentation

Ohya

Ohyama

Kobayashi

2005

Biotech & Bioengineering

View full text Add to dashboard Cite

An optimization method for repeated fed-batch fermentation was established with the aim of improving the recombinant human serum albumin (rHSA) production in Pichia pastoris. A simulation model for fed-batch fermentation was formulated and the optimal methanol-feeding policy calculated by dynamic programming method using five different methanol-feeding periods. The necessary state variables were collected from the calculated results and used for further optimization of repeated fed-batch fermentation. The optimal operation policy was investigated using the pre-collected state variables by estimating the overall profit per total methanol-feeding time. The calculated results indicated that the initial cell mass from the 2nd fed-batch fermentation on should be set at 35 or 40 g and methanol-feeding time at 264 h. In repeated fed-batch fermentation using the optimal operation policy, actual culture volume was in good agreement with the values simulated by model equations, but some discrepancy was observed in rHSA production. Minimum experiments were therefore carried out to re-evaluate rHSA production levels, which were then applied in re-calculations to determine the optimal operation policy. The optimal policy for repeated fed-batch fermentation established in the present study (i.e., 4-times-repeated fed-batch fermentation) achieved a 47% increase in annual rHSA production. Optimization of the culture period also brought about a 28% increase in annual rHSA production even in simple (not repeated) fed-batch fermentation.

show abstract

Section: Introductionmentioning

confidence: 99%

Optimization of human serum albumin production in methylotrophic yeast Pichia pastoris by repeated fed‐batch fermentation

Ohya

Ohyama

Kobayashi

2005

Biotech & Bioengineering

View full text Add to dashboard Cite

show abstract

“…The constitutive promoters most commonly used in this yeast are derived from the phosphoglycerate kinase (PGK ), alcohol dehydrogenase (ADH ) and glyceraldehyde-3-phosphate dehydrogenase (GAP ) genes. The inducible promoters commonly used are derived from the galactose-regulated genes GAL1, GAL7 and GAL10 or from the phosphate-regulated gene PHO5, which is induced in response to phosphate starvation (Mendoza-Vega et al, 1994;Schneider and Guarente, 1991).…”

Section: Introductionmentioning

confidence: 99%

The ICL1 gene of Pichia pastoris, transcriptional regulation and use of its promoter

Menéndez¹,

Valdés²,

Cabrera³

2003

Yeast

View full text Add to dashboard Cite

We cloned and characterized a gene encoding isocitrate lyase from the methylotrophic yeast Pichia pastoris. This gene was isolated from a P. pastoris genomic library using a homologous PCR hybridization probe, amplified with two sets of degenerate primers designed from conserved regions in yeast isocitrate lyases. The cloned gene was sequenced and consists of an open reading frame of 1563 bp encoding a protein of 551 amino acids. The molecular mass of the protein is calculated to be 60.6 kDa with high sequence similarity to isocitrate lyase from other organisms. There is a 64% identity between amino acid sequences of P. pastoris Icl and Saccharomyces cerevisiae Icl. Northern blot analyses showed that, as in S. cerevisiae, the steady-state ICL1 mRNA levels depend on the carbon source used for cell growth. Expression in P. pastoris of the dextranase gene (dexA) from Penicillium minioluteum under control of the ICL1 promoter proved that P ICL1 is a good alternative for the expression of heterologous proteins in this methylotrophic yeast. The sequence presented here has been deposited in the EMBL data library under Accession No. AJ272040.

show abstract

“…This organism is traditionally used for large-scale production of baker's yeast and ethanol, with a considerable increase in the production of fuel ethanol in the last 3 decades (4,6,50). Additionally, this platform has been successfully applied to the production of valuable heterologous proteins on an industrial scale (26,35), such as various FDA-approved pharmaceuticals, including insulin (34) and hepatitis B surface antigen (23). Yeast-based expression systems excel because of their available constitutive or strongly inducible promoters and their growth to high cell densities on inexpensive substrates.…”

mentioning

confidence: 99%

Synthesis and Accumulation of Cyanophycin in Transgenic Strains of Saccharomyces cerevisiae

Steinle

Oppermann‐Sanio

Reichelt

et al. 2008

Appl Environ Microbiol

View full text Add to dashboard Cite

Cyanophycin [multi-L-arginyl-poly(L-aspartic acid) (CGP)] was, for the first time, produced in yeast. As yeasts are very important production organisms in biotechnology, it was determined if CGP can be produced in two different strains of Saccharomyces cerevisiae. The episomal vector systems pESC (with the galactoseinducible promoter GAL1) and pYEX-BX (with the copper ion-inducible promoter CUP1) were chosen to express the cyanophycin synthetase gene from the cyanobacterium Synechocystis sp. strain PCC 6308 (cphA 6308 ) in yeast. Expression experiments with transgenic yeasts revealed that the use of the CUP1 promoter is much more efficient for CGP production than the GAL1 promoter. As observed by electrophoresis of isolated CGP in sodium dodecyl sulfate-polyacrylamide gels, the yeast strains produced two different types of polymer: the water-soluble and the water-insoluble CGP were observed as major and minor forms of the polymer, respectively. A maximum CGP content of 6.9% (wt/wt) was detected in the cells. High-performance liquid chromatography analysis showed that the isolated polymers consisted mainly of the two amino acids aspartic acid and arginine and that, in addition, a minor amount (2 mol%) of lysine was present. Growth of transgenic yeasts in the presence of 15 mM lysine resulted in an incorporation of up to 10 mol% of lysine into CGP. Anti-CGP antibodies generated against CGP isolated from Escherichia coli TOP10 harboring cphA 6308 reacted with insoluble CGP but not with soluble CGP, if applied in Western or dot blots.Cyanophycin, also referred to as multi-L-arginyl-poly(L-aspartic acid) or cyanophycin granule polypeptide (CGP), is a nonribosomally synthesized polypeptide consisting of a poly-(aspartic acid) backbone with arginine residues linked to the ␤-carboxyl group of each aspartate by the ␣-amino group (45). CGP is a polydispersed polymer; the molecular size distribution of CGP varies with the producing host strains (1,15,18,20,30,37,52). Due to its branched structure, CGP is not degradable by a wide range of proteinases (45). Biosynthesis of CGP from aspartate and arginine requires only one enzyme, cyanophycin synthetase, which is encoded by cphA (51). CGP is insoluble at neutral pH and under physiological ionic strength, but it is soluble at low (Ͼ3) or high (Ͻ9) pH. Ziegler et al. were the first to observe a water-soluble form of CGP after the heterologous expression of cphA from Desulfitobacterium hafniense strain DSM 10664 in Escherichia coli (52). A detailed study of the solubility behavior of CGP isolated from recombinant E. coli in inorganic salts has been carried out by Füser and Steinbüchel (16). It was shown that the occurrence of the soluble form was not dependent on the origin of cphA or on the host.Recently, several putative applications for CGP and its derivatives have become available, indicating there is a need for its efficient biotechnological production (36,42,43). For the production of CGP at a technical scale, cyanobacteria were shown to be unsuitable due to their low cell...

show abstract

Industrial production of heterologous proteins by fed-batch cultures of the yeast Saccharomyces cereuisiae

Cited by 27 publications

References 113 publications

Optimization of human serum albumin production in methylotrophic yeast Pichia pastoris by repeated fed‐batch fermentation

Optimization of human serum albumin production in methylotrophic yeast Pichia pastoris by repeated fed‐batch fermentation

The ICL1 gene of Pichia pastoris, transcriptional regulation and use of its promoter

Synthesis and Accumulation of Cyanophycin in Transgenic Strains of Saccharomyces cerevisiae

Contact Info

Product

Resources

About