Mixed feeds of glycerol and methanol can improve the performance of Pichia pastoris cultures: A quantitative study based on concentration gradients in transient continuous cultures

Jungo, Carmen; Marison, I. W.; Stockar, Urs von

doi:10.1016/j.jbiotec.2006.12.024

Cited by 103 publications

(90 citation statements)

References 36 publications

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“…If on-line heat monitoring is added as well, the resulting four on-line rate measurements may be combined with typically five constraints resulting from balances for carbon, degree of reduction, nitrogen, charge, and energy to yield a considerably over-determined set of rates, at least for typical microbial bioprocess stoichiometry. It is therefore possible to check for gross measurement errors, to reconcile the data for more probable values, and to determine the unmeasured rates, including the growth rate, on line [41,42].…”

Section: On-line Monitoring and Control Of Bioprocesses By Heat Dissimentioning

confidence: 99%

Biothermodynamics of live cells: a tool for biotechnology and biochemical engineering

Stockar

2010

Journal of Non-Equilibrium Thermodynamics

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The aim of this contribution is to review the application of thermodynamics to live cultures of microbial and other cells and to explore to what extent this may be put to practical use. A major focus is on energy dissipation effects in industrially relevant cultures, both in terms of heat and Gibbs energy dissipation. The experimental techniques for calorimetric measurements in live cultures are reviewed and their use for monitoring and control is discussed. A detailed analysis of the dissipation of Gibbs energy in chemotrophic growth shows that it reflects the entropy production by metabolic processes in the cells and thus also the driving force for growth and metabolism. By splitting metabolism conceptually up into catabolism and biosynthesis, it can be shown that this driving force decreases as the growth yield increases. This relationship is demonstrated by using experimental measurements on a variety of microbial strains. On the basis of these data, several literature correlations were tested as tools for biomass yield prediction. The prediction of other culture performance characteristics, including product yields for biorefinery planning, energy yields for biofuel manufacture, maximum growth rates, maintenance requirements, and threshold concentrations is also briefly reviewed.

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Section: On-line Monitoring and Control Of Bioprocesses By Heat Dissimentioning

confidence: 99%

Biothermodynamics of live cells: a tool for biotechnology and biochemical engineering

Stockar

2010

Journal of Non-Equilibrium Thermodynamics

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“…Scenarios B1-B7 and C1-C2 correspond to cultures producing a lipase from Rhizopus oryzae (ROL) [31,32]. Scenarios D1-D10 have been taken from P. pastoris cultures expressing and secreting recombinant avidin [33]. Scenario E1 has been obtained from a macrokinetic model for P. pastoris expressing recombinant human serum albumin (HSA) [34].…”

Section: P Pastoris Experimental Data Setmentioning

confidence: 99%

MCR-ALS on metabolic networks: Obtaining more meaningful pathways

Folch-Fortuny

Tortajada

Prats-Montalbán

et al. 2015

Chemometrics and Intelligent Laboratory Systems

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With the aim of understanding the flux distributions across a metabolic network, i.e. within living cells, Principal Component Analysis (PCA) has been proposed to obtain a set of orthogonal components (pathways) capturing most of the variance in the flux data. The problems with this method are (i) that no additional information can be included in the model, and (ii) that orthogonality imposes a hard constraint, not always reasonably. To overcome these drawbacks, here we propose to use a more flexible approach such as Multivariate Curve Resolution -Alternating Least Squares (MCR-ALS) to obtain this set of biological pathways through the network. By using this method, different constraints can be included in the model, and the same source of variability can be present in different pathways, which is reasonable from a biological standpoint. This work follows a methodology developed for Pichia pastoris cultures grown on different carbon sources, lately presented in [González Martínez et al. 2014].In this paper a different grey modelling approach, which aims to incorporate a priori knowledge through constraints on the modelling algorithms, is applied to the same case of study. The results of both models are compared to show their strengths and weaknesses.

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“…Also, methanol accumulation in the fermenter is toxic to recombinant P. pastoris and can have a negative influence on protein induction (Jungo et al 2007a). Therefore, the combined glycerol and methanol feed strategy is one of the most important factors for maximizing heterologous protein production (Cos et al 2006).…”

Section: Introductionmentioning

confidence: 99%

Combined Strategies for Improving the Production of Recombinant Rhizopus oryzae Lipase in Pichia pastoris

Li¹,

He²,

Li³

et al. 2013

BioResources

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We have developed a yeast Pichia pastoris system for the high-level expression of recombinant Rhizopus oryzae lipase (ROL), which is a potentially effective catalyst in the solvent-free production of biodiesel fuel. In the glycerol fed-batch phase, the combination of the dissolvedoxygen-stat and gradient-control glycerol feeding strategies resulted in a higher cell biomass in the P. pastoris culture, with shorter feed times. In the methanol fed-batch phase, a constant methanol concentration of 0.3-0.5% (v/v) was found to be optimal for high ROL activity, maximum protein concentration, and a maximum biomass. Cells were grown at 30 °C and induced by methanol at 22 °C and a pH of 6.0 with the addition of 0.5% (w/v) casein, which reduced the proteolytic degradation of ROL. Using a combined glycerol step-increasing, on-line methanol feeding strategy and these reducing proteolytic degradation methods, a maximum biomass of 96 g DCW/l, a maximum concentration of 2.0 g/L, and an activity of 1302.2 U/mL ROL were attained. The simple culture process is the highest level of ROL expression reported in P. pastoris. Our results strongly demonstrate that the efficiency of the recombinant ROL expression is directly dependent on feed strategies, lower induction temperature, maintaining the proper pH, and the addition of casein.

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Mixed feeds of glycerol and methanol can improve the performance of Pichia pastoris cultures: A quantitative study based on concentration gradients in transient continuous cultures

Cited by 103 publications

References 36 publications

Biothermodynamics of live cells: a tool for biotechnology and biochemical engineering

Biothermodynamics of live cells: a tool for biotechnology and biochemical engineering

MCR-ALS on metabolic networks: Obtaining more meaningful pathways

Combined Strategies for Improving the Production of Recombinant Rhizopus oryzae Lipase in Pichia pastoris

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