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      Applicability of a single‐use bioreactor compared to a glass bioreactor for the fermentation of filamentous fungi and evaluation of the reproducibility of growth in pellet form

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          Abstract

          The implementation of single‐use technologies offers several major advantages, e.g. prevention of cross‐contamination, especially when spore‐forming microorganisms are present. This study investigated the application of a single‐use bioreactor in batch fermentation of filamentous fungus Penicillium sp. (IBWF 040‐09) from the Institute of Biotechnology and Drug Research (IBWF), which is capable of intracellular production of a protease inhibitor against parasitic proteases as a secondary metabolite. Several modifications to the SU bioreactor were suggested in this study to allow the fermentation in which the fungus forms pellets. Simultaneously, fermentations in conventional glass bioreactor were also conducted as reference. Although there are significant differences in the construction material and gassing system, the similarity of the two types of bioreactors in terms of fungal metabolic activity and the reproducibility of fermentations could be demonstrated using statistic methods. Under the selected cultivation conditions, growth rate, yield coefficient, substrate uptake rate, and formation of intracellular protease‐inhibiting substance in the single‐use bioreactor were similar to those in the glass bioreactor.

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          Most cited references41

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          Single-use disposable technologies for biopharmaceutical manufacturing.

          The manufacture of protein biopharmaceuticals is conducted under current good manufacturing practice (cGMP) and involves multiple unit operations for upstream production and downstream purification. Until recently, production facilities relied on the use of relatively inflexible, hard-piped equipment including large stainless steel bioreactors and tanks to hold product intermediates and buffers. However, there is an increasing trend towards the adoption of single-use technologies across the manufacturing process. Technical advances have now made an end-to-end single-use manufacturing facility possible, but several aspects of single-use technology require further improvement and are continually evolving. This article provides a perspective on the current state-of-the-art in single-use technologies and highlights trends that will improve performance and increase the market penetration of disposable manufacturing in the future.
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            The filamentous fungal pellet and forces driving its formation

            Filamentous fungi play an important role not only in the bio-manufacturing of value-added products, but also in bioenergy and environmental research. The bioprocess manipulation of filamentous fungi is more difficult than that of other microbial species because of their different pellet morphologies and the presence of tangled mycelia under different cultivation conditions. Fungal pellets, which have the advantages of harvest ease, low fermentation broth viscosity and high yield of some proteins, have been used for a long time. Many attempts have been made to establish the relationship between pellet and product yield using quantitative approaches. Fungal pellet formation is attributed to the combination of electrostatic interactions, hydrophobicity and specific interactions from spore wall components. Electrostatic interactions result from van der Waals forces and negative charge repulsion from carboxyl groups in the spore wall structure. Electrostatic interactions are also affected by counter-ions (cations) and the physiologic conditions of spores that modify the carboxyl groups. Fungal aggregates are promoted by the hydrophobicity generated by hydrophobins, which form a hydrophobic coat that covers the spore. The specific interactions of spore wall components contribute to spore aggregation through salt bridging. A model of spore aggregation was proposed based on these forces. Additionally, some challenges were addressed, including the limitations of research techniques, the quantitative determination of forces and the complex information of biological systems, to clarify the mechanism of fungal pellet formation.
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              Morphology and productivity of filamentous fungi.

              Cultivation processes involving filamentous fungi have been optimised for decades to obtain high product yields. Several bulk chemicals like citric acid and penicillin are produced this way. A simple adaptation of cultivation parameters for new production processes is not possible though. Models explaining the correlation between process-dependent growth behaviour and productivity are therefore necessary to prevent long-lasting empiric test series. Yet, filamentous growth consists of a complex microscopic differentiation process from conidia to hyphae resulting in various macroscopically visible appearances. Early approaches to model this morphologic development are recapitulated in this review to explain current trends in this area of research. Tailoring morphology by adjusting process parameters is one side of the coin, but an ideal morphology has not even been found. This article reviews several reasons for this fact starting with nutrient supply in a fungal culture and presents recent advances in the investigation of fungal metabolism. It illustrates the challenge to unfold the relationship between morphology and productivity.
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                Author and article information

                Contributors
                w.soerjawinata@umwelt-campus.de
                Journal
                Eng Life Sci
                Eng Life Sci
                10.1002/(ISSN)1618-2863
                ELSC
                Engineering in Life Sciences
                John Wiley and Sons Inc. (Hoboken )
                1618-0240
                1618-2863
                25 February 2021
                May 2021
                : 21
                : 5 ( doiID: 10.1002/elsc.v21.5 )
                : 324-339
                Affiliations
                [ 1 ] Institute for Biotechnical Process Design Trier University of Applied Sciences, Environmental Campus Birkenfeld Hoppstädten‐Weiersbach Germany
                [ 2 ] Institute of Pharmaceutical and Biomedical Sciences Johannes Gutenberg University Mainz Mainz Germany
                [ 3 ] Institut für Biotechnologie und Wirkstoff‐Forschung gGmbH (IBWF) Mainz Germany
                [ 4 ] Institute of Bioprocess Engineering Technical University Kaiserslautern Kaiserslautern Germany
                Author notes
                [*] [* ] Correspondence

                Winda Soerjawinata, MSc, Institute for Biotechnical Process Design, Trier University of Applied Science, Environmental Campus Birkenfeld, Campusallee 9913, 55768 Hoppstädten‐Weiersbach, Germany.

                Email: w.soerjawinata@ 123456umwelt-campus.de

                Author information
                https://orcid.org/0000-0002-7674-0967
                Article
                ELSC1373
                10.1002/elsc.202000069
                8092982
                33976605
                ce6bb2a9-b8f8-432f-8d60-d676428c5a8a
                © 2021 The Authors. Engineering in Life Sciences published by Wiley‐VCH GmbH

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 22 January 2021
                : 04 September 2020
                : 12 February 2021
                Page count
                Figures: 8, Tables: 1, Pages: 16, Words: 9905
                Categories
                Research Article
                Research Articles
                Custom metadata
                2.0
                May 2021
                Converter:WILEY_ML3GV2_TO_JATSPMC version:6.0.2 mode:remove_FC converted:03.05.2021

                fermentation,pellets,penicillium sp,protease inhibitor,single‐use bioreactor

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