A new stoichiometric miniaturization strategy for screening of industrial microbial strains: application to cellulase hyper-producing Trichoderma reesei strains

Microbial Cell Factories

Research

Christian Larroche¹  Laurent Poughon¹  Fadhel Ben Chaabane²  Etienne Jourdier²  Frédéric Monot² 

[1] Clermont Université, Université Blaise Pascal, Institut Pascal, Polytech Clermont-Ferrand, 24 av. des Landais, BP 20206, 63174, Aubière cedex, France;IFP Energies nouvelles, 1 et 4 avenue de Bois-Préau, 92852, Rueil-Malmaison, France;

关键词: Screening;    Miniaturization;    Stoichiometry;    Fed-batch;    pH Control;    Cellulase;    Trichoderma reesei;   

DOI  :  10.1186/1475-2859-11-70

 received in 2012-03-22, accepted in 2012-05-30,  发布年份 2012

来源: Springer

【 摘 要 】

BackgroundDuring bioprocess development, secondary screening is a key step at the boundary between laboratory and industrial conditions. To ensure an effective high-throughput screening, miniaturized laboratory conditions must mimic industrial conditions, especially for oxygen transfer, feeding capacity and pH stabilization.ResultsA feeding strategy has been applied to develop a simple screening procedure, in which a stoichiometric study is combined with a standard miniaturization procedure. Actually, the knowledge of all nutriments and base or acid requirements leads to a great simplification of pH stabilization issue of miniaturized fed-batch cultures. Applied to cellulase production by Trichoderma reesei, this strategy resulted in a stoichiometric mixed feed of carbon and nitrogen sources. While keeping the pH between shake flask and stirred bioreactor comparable, the developed shake flask protocol reproduced the strain behaviour under stirred bioreactor conditions. Compared to a an already existing miniaturized shake flasks protocol, the cellulase concentration was increased 5-fold, reaching about 10 g L-1. Applied to the secondary screening of several clones, the newly developed protocol succeeded in selecting a clone with a high industrial potential.ConclusionsThe understanding of a bioprocess stoichiometry contributed to define a simpler and more effective miniaturization. The suggested strategy can potentially be applied to other fed-batch processes, for the screening of either strain collections or experimental conditions.

【 授权许可】

Unknown   
© Jourdier et al.; licensee BioMed Central Ltd. 2012. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.