The stability of the yeast plasmid pJDB248 depends on growth rate of the culture

Kleinman, Matthew J.; Gingold, Elliot B.; Stanbury, Peter F.

doi:10.1007/bf01030502

Cited by 33 publications

(17 citation statements)

References 10 publications

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“…Differences in plasmid stability of recombinant plasmids in yeast strains have also been observed by Mead et al (1986) and Lee et al (1986), although they analysed only a small number of strains. Kleinman et al (1986) and Parker and DiBiasio (1987) observed a correlation between plasmid stability, growth rate, and the medium employed. Initial findings in recombinant systems have shown the deleterious effects of low oxygen tension on plasmid stability (Hopkins et al 1987) and it is therefore likely that recombinant yeast will be sensitive to local changes in oxygen tension that can occur in large-scale fermentors during the growth a n d / o r production phase.…”

Section: Discussionmentioning

confidence: 99%

Expression of the human blood coagulation protein Factor XIIIa in Saccharomyces cerevisiae: dependence of the expression levels from host-vector systems and medium conditions

Bröker

Bäuml

Göttig

et al. 1991

Appl Microbiol Biotechnol

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The human blood coagulation protein Factor XIIIa (FXIIIa) was expressed in Saccharomyces cerevisiae employing Escherichia coli-yeast shuttle vectors based on a 2-mu plasmid. Several factors affecting high production yield of recombinant FXIIIa were analysed. The use of the regulatable GAL-CYC1 hybrid promoter resulted in higher FXIIIa expression when compared with the constitutive ADCI promoter. Screening for suitable yeast strains for expression of FXIIIa under the transcriptional control of the GAL-CYC1 hybrid promoter revealed a broad spectrum of productivity. No obvious correlation between the expression rate and the genetic markers of the strains could be identified. The medium composition markedly influenced the FXIIIa expression rates. The expression of FXIIIa was strictly regulated by the carbon source. Glucose as the only sugar and energy source repressed the synthesis of FXIIIa, whereas addition of galactose induced FXIIIa expression. Special feeding schemes resulted in a productivity of up to 100 mg FXIIIa/l in shake flasks.

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Section: Discussionmentioning

confidence: 99%

Expression of the human blood coagulation protein Factor XIIIa in Saccharomyces cerevisiae: dependence of the expression levels from host-vector systems and medium conditions

Bröker

Bäuml

Göttig

et al. 1991

Appl Microbiol Biotechnol

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“…The reason for the decrease in production activity of cells has been explained by means of several properties of plasmid and gene expression (Kleinman et al 1986;Parker and DiBiasio 1987). According to our experience, the activity of a recombinant S. cerevisiae producing HBsAg decreased below 50% of the m a x i m u m yield Maintenance of constant specific growth rate in fed-batch culture at generation numbers 10 to 15 (flask repeat number 3-4).…”

Section: Batch Cultivationmentioning

confidence: 99%

Growth rate control in fed-batch cultures of recombinant Saccharomyces cerevisiae producing hepatitis B surface antigen (HBsAg)

Gu¹,

Park²,

Kim³

1991

Appl Microbiol Biotechnol

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A recombinant Saccharomyces cerevisiae producing hepatitis B surface antigen (HBsAg) exhibited growth-associated product formation. By controlling the medium feed rate, based on the calculated amount of medium required for 1 h, a constant specific growth rate was obtained in the range of 0.12-0.18 h-1. In order to prolong the exponential growth phase, the medium feed rate was increased exponentially. A fed-batch cultivation method based on the production kinetics of batch culture enhanced HBsAg production ten times more than in batch culture. The reason for the increase can be explained by the fact that the production of HBsAg is expressed as an exponential function of time when the specific growth rate is controlled to a constant value in growth-associated product formation kinetics. In the scale-up of this culture to 91, the specific growth rate could also be maintained constant and the HBsAg production trend was similar to that in a 1-1 culture. However, ethanol accumulation occurred at a late stage in fed-bach culture. Ethanol produced was not reutilized and inhibited further cell growth.

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“…cerevisiae, the maintenance of 2p and 2p-based chimeric plasmids in cell populations has been investigated during batch growth (Futcher & Cox, 1984) and in continuous cultures (Walmsley et al, 1983;Mead et al,1986a, b;Kleinman et al, 1986). The behaviour of different plasmids has been found to vary considerably and factors influencing stability have included t Present address: School of Biological Sciences, Queen Mary College, London, UK.…”

Section: Introductionmentioning

confidence: 99%

The Segregation of the 2 -based Yeast Plasmid pJDB248 Breaks Down under Conditions of Slow, Glucose-limited Growth

et al. 1989

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The stability of the 2 mu-based yeast plasmid pJDB248 in Saccharomyces cerevisiae S150-2B(cir0) was investigated in glucose-limited chemostat culture. Plasmid-free cells were detected by loss of (plasmid-encoded) leucine prototrophy and confirmed by colony hybridization. The plasmid was considerably more stable at a high dilution rate (0.12 h-1) than at a lower dilution rate (0.05 h-1). The average plasmid copy number in the cells retaining the plasmid remained constant at approximately 50 in the high dilution rate culture whereas it rose to almost 600 in the slow dilution rate culture. However, in both cultures the overall plasmid level in the total population remained constant, indicating that plasmid segregation breaks down at the low growth rate. Similar experiments on the native 2 mu plasmid demonstrated high stability and no significant differences between the high and low growth rate cultures. It is postulated that the difference in behaviour between the native and chimeric plasmids is related to an interaction between the growth conditions and the loss of the D gene product.

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The stability of the yeast plasmid pJDB248 depends on growth rate of the culture

Cited by 33 publications

References 10 publications

Expression of the human blood coagulation protein Factor XIIIa in Saccharomyces cerevisiae: dependence of the expression levels from host-vector systems and medium conditions

Expression of the human blood coagulation protein Factor XIIIa in Saccharomyces cerevisiae: dependence of the expression levels from host-vector systems and medium conditions

Growth rate control in fed-batch cultures of recombinant Saccharomyces cerevisiae producing hepatitis B surface antigen (HBsAg)

The Segregation of the 2 -based Yeast Plasmid pJDB248 Breaks Down under Conditions of Slow, Glucose-limited Growth

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