Optimisation of culture conditions with respect to biotin requirement for the production of recombinant avidin in Pichia pastoris

Jungo, Carmen; Urfer, Julien; Zocchi, Andrea; Marison, I. W.; Stockar, Urs von

doi:10.1016/j.jbiotec.2006.08.001

Cited by 20 publications

(11 citation statements)

References 32 publications

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“…Various strategies were tested in order to decrease Q blocked . Cells neither grew in biotin‐free BSM medium, nor in a medium in which biotin was substituted by oleic acid and aspartic acid as described for the production of avidin by P. pastoris (data not shown) . This may be the result of the carbon source glucose, which seems to inhibit the positive effects of the substitutes .…”

Section: Resultssupporting

confidence: 53%

“…Cells neither grew in biotin-free BSM medium, nor in a medium in which biotin was substituted by oleic acid and aspartic acid as described for the production of avidin by P. pastoris (data not shown). 57 This may be the result of the carbon source glucose, which seems to inhibit the positive effects of the substitutes. 37 Replacement of biotin by small concentrations of complex supplements (soy peptone and yeast extract, 2 g L 21 each) allowed the accumulation of only small concentrations of both SAV and biomass.…”

Section: Bioreactor Experimentsmentioning

confidence: 99%

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GAP promoter‐based fed‐batch production of highly bioactive core streptavidin by Pichia pastoris

Müller

Bruhn

Flaschel

et al. 2016

Biotechnology Progress

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Streptavidin is a homotetrameric protein binding the vitamin biotin and peptide analogues with an extremely high affinity, which leads to a large variety of applications. The biotin-auxotrophic yeast Pichia pastoris has recently been identified as a suitable host for the expression of the streptavidin gene, allowing both high product concentrations and productivities. However, so far only methanol-based expression systems have been applied, bringing about increased oxygen demand, strong heat evolution and high requirements for process safety, causing increased cost. Moreover, common methanol-based processes lead to large proportions of biotin-blocked binding sites of streptavidin due to biotin-supplemented media. Targeting these problems, this paper provides strategies for the methanol-free production of highly bioactive core streptavidin by P. pastoris under control of the constitutive GAP promoter. Complex were superior to synthetic production media regarding the proportion of biotin-blocked streptavidin. The optimized, easily scalable fed-batch process led to a tetrameric product concentration of up to 4.16 ± 0.11 µM of biotin-free streptavidin and a productivity of 57.8 nM h(-1) based on constant glucose feeding and a successive shift of temperature and pH throughout the cultivation, surpassing the concentration in un-optimized conditions by a factor of 3.4. Parameter estimation indicates that the optimized conditions caused a strongly increased accumulation of product at diminishing specific growth rates (μ ≈ D < 0.01 h(-1) ), supporting the strategy of feeding. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:855-864, 2016.

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Section: Resultssupporting

confidence: 53%

Section: Bioreactor Experimentsmentioning

confidence: 99%

GAP promoter‐based fed‐batch production of highly bioactive core streptavidin by Pichia pastoris

Müller

Bruhn

Flaschel

et al. 2016

Biotechnology Progress

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“…In our experiment, no medium exchange was carried out, and we suggest that salt concentration may not affect a Mut + strain. Biotin has been reported as essential to growth rate and biomass yield for some proteins [28]. Different amounts of biotin were tested in our experiments, and we found that yeast growth remained unaffected.…”

Section: Discussionmentioning

confidence: 57%

Overproduction of 15N-Labeled r-RGD-Hirudin in Pichia pastoris for NMR Studies

Wang

Zhang²,

Li³

et al. 2010

PPL

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The novel recombinant hirudin, r-RGD-hirudin, inhibits thrombin and platelet aggregation. Here, we reported over-expression of (15)N-labeled r-RGD-hirudin by Pichia pastoris in minimal medium. After extensive optimization, the yield of active r-RGD-hirudin reached ≈600 mg/L when the yeast cells were culture in a fermenter. The purified (15)N-labeled r-RGD-hirudin retained full biological activity and was uniformly labeled. Heteronuclear NMR of the (15)N-labeled r-RGD-hirudin was performed for the first time, and all signals in the heteronuclear single quantum coherence (HSQC) spectrum were successfully assigned.

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“…Recently, a growing number of studies have succeeded in monitoring the heat dissipation of microbial during fermentation by ITC [33][34][35][36][37]. Kemp and his colleagues [23,24] had combined a bioreactor with a flow microcalorimeter to monitor the heat flow rate of the growth of animal cells under the controlled conditions for long cultivation time.…”

Section: Resultsmentioning

confidence: 99%

Using isothermal titration calorimetry to real-time monitor the heat of metabolism: A case study using PC12 cells and Aβ(1–40)

Wang

Lin

Chen

et al. 2011

Colloids and Surfaces B: Biointerfaces

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Optimisation of culture conditions with respect to biotin requirement for the production of recombinant avidin in Pichia pastoris

Cited by 20 publications

References 32 publications

GAP promoter‐based fed‐batch production of highly bioactive core streptavidin by Pichia pastoris

GAP promoter‐based fed‐batch production of highly bioactive core streptavidin by Pichia pastoris

Overproduction of 15N-Labeled r-RGD-Hirudin in Pichia pastoris for NMR Studies

Using isothermal titration calorimetry to real-time monitor the heat of metabolism: A case study using PC12 cells and Aβ(1–40)

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