Comparative study on the production of guar alpha-galactosidase by Saccharomyces cerevisiae SU50B and Hansenula polymorpha 8/2 in continuous cultures

Giuseppin, Marco L. F.; Almkerk, José W. van; Heistek, Jolanda C.; Verrips, C. Theo

doi:10.1128/aem.59.1.52-59.1993

Cited by 39 publications

(16 citation statements)

References 23 publications

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“…Otherwise, I assumed that 1% of product uptake would be allocated to enzyme production with the remainder stored in biomass. This value is within the range of 0.7-2.1% reported for a-glucosidase production by yeasts in continuous cultures (Giuseppin et al 1993) and slightly higher than the 0.3-0.9% reported for protease production by Bacillus clausii (Christiansen & Nielsen 2002). I also assumed (conservatively) that a quantity of C equal to 10% of enzyme C mass would be respired to account for additional metabolic costs of enzyme production.…”

Section: Uptake and Enzyme Productionmentioning

confidence: 72%

Cheaters, diffusion and nutrients constrain decomposition by microbial enzymes in spatially structured environments

2005

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Extracellular enzymes allow microbes to acquire carbon and nutrients from complex molecules and catalyse the rate-limiting step in nutrient mineralization. Because the factors regulating enzyme production are poorly understood, I used a simulation model to examine how competition, nutrient availability and spatial structure affect microbial growth and enzyme synthesis. In simulations where enzyme-producing microbes competed with cheaters (who do not synthesize enzymes but take-up product), higher enzyme costs favoured cheaters, while lower rates of enzyme diffusion favoured producers. Cheaters and producers coexisted in highly organized spatial patterns at intermediate enzyme costs and diffusion rates. Simulations with varying nutrient inputs showed that nitrogen supply can limit carbon mineralization, microbial growth and enzyme production because of the nitrogen-demanding stoichiometry of enzymes (C : N ¼ c. 3.5 : 1). These results suggest that competition from cheaters, slow diffusion and nitrogen limitation may constrain microbial foraging and the enzymatic decomposition of complex compounds in natural environments.

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Section: Uptake and Enzyme Productionmentioning

confidence: 72%

Cheaters, diffusion and nutrients constrain decomposition by microbial enzymes in spatially structured environments

2005

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“…The maximum specific growth rate (l max ) of recombinant P. stipitis PJH53 during fully aerobic growth (0.15 h À1 ) was significantly lower than previously reported for wild-type P. stipitis strains under similar conditions (0.27-0.38 h À1 [29,30]). In recombinant strains of S. cerevisiae such dramatic decrease in the l max , compared to parental strains, is not routinely reported, especially not in the absence of heterologous gene expression [ [31][32][33][34][35]. It has been demonstrated that the mutant strain PJH53 grew slightly slower than its corresponding wildtype strain CBS5774 under all aeration conditions, probably due to an unknown mutation of the strain [30].…”

Section: Resultsmentioning

confidence: 99%

Amino acid supplementation, controlled oxygen limitation and sequential double induction improves heterologous xylanase production by

Görgens

Passoth

Zyl

et al. 2005

FEMS Yeast Research

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Heterologous endo-beta-1,4-xylanase was produced by Pichia stipitis under control of the hypoxia-inducible PsADH2-promoter in a high-cell-density culture. After promoter induction by a shift to oxygen limitation, different aeration rates (oxygen transfer rates) were applied while maintaining oxygen-limitation. Initially, enzyme production was higher in oxygen-limited cultures with high rates of oxygen transfer, although the maximum xylanase activity was not significantly influenced. Amino acid supplementation increased the production of the heterologous endo-beta-1,4-xylanase significantly in highly aerated oxygen-limited cultures, until glucose was depleted. A slight second induction of the promoter was observed in all cultures after the glucose had been consumed. The second induction was most obvious in amino acid-supplemented cultures with higher oxygen transfer rates during oxygen limitation. When such oxygen-limited cultures were shifted back to fully aerobic conditions, a significant re-induction of heterologous endo-beta-1,4-xylanase production was observed. Re-induction was accompanied by ethanol consumption. A similar protein production pattern was observed when cultures were first grown on ethanol as sole carbon source and subsequently glucose and oxygen limitation were applied. Thus, we present the first expression system in yeast with a sequential double-inducible promoter.

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“…The growth rate in the exponential feed phase is of major importance for the productivity of the fermentation (Giuseppin et al, 1993). The glucose and ethanol limited fed-batch fermentations were performed with a growth rate of 0.06 h À1 in the exponential phase as that proved to be the best growth rate for optimal production of heterologous proteins.…”

Section: Carbon Limited Exponential Growthmentioning

confidence: 99%

Increased heterologous protein production by Saccharomyces cerevisiae growing on ethanol as sole carbon source

Laar¹,

Visser²,

Holster³

et al. 2006

Biotech & Bioengineering

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Saccharomyces cerevisiae is a widely used host organism for the production of heterologous proteins, often cultivated in glucose-based fed-batch processes. This production system however has many factors limiting the productivity, mainly towards the end of the fermentation. For the optimised production of a Camelid antibody fragment this process was evaluated. In shake flask cultivations, it was found that ethanol has a strong effect on productivity increase and therefore glucose and ethanol fed-batch fermentations were compared. It appeared that specific heterologous protein production was up to five times higher in the ethanol cultivation and could be further optimised. Then the key characteristics of ethanol fed-batch fermentations such as growth rate and specific production were determined under ethanol limitation and accumulation and growth limiting conditions in the final phase of the process. It appeared that an optimal production process should have an ethanol accumulation throughout the feed phase of approximately 1% v/v in the broth and that production remains very efficient even in the last phase of the process. This productivity increase on ethanol versus glucose was also proven for several other Camelid antibody fragments some of which were heavily impaired in secretion on glucose, but very well produced on ethanol. This leads to the suggestion that the ethanol effect on improved heterologous protein production is linked to a stress response and folding and secretion efficiency.

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Comparative study on the production of guar alpha-galactosidase by Saccharomyces cerevisiae SU50B and Hansenula polymorpha 8/2 in continuous cultures

Cited by 39 publications

References 23 publications

Cheaters, diffusion and nutrients constrain decomposition by microbial enzymes in spatially structured environments

Cheaters, diffusion and nutrients constrain decomposition by microbial enzymes in spatially structured environments

Amino acid supplementation, controlled oxygen limitation and sequential double induction improves heterologous xylanase production by

Increased heterologous protein production by Saccharomyces cerevisiae growing on ethanol as sole carbon source

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