+1 Recommend
0 collections
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      A microfluidic device for simultaneous detection of enzyme secretion and elongation of a single hypha


      Read this article at

          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.


          Filamentous fungi grow through elongation of their apical region by exocytosis and secrete enzymes that can be of commercial or industrial importance. Their hyphae exhibit extensive branching, making it difficult to control hyphal growth for observation and analysis. Therefore, although hyphal morphology and productivity are closely related, the relationship between the two has not yet been clarified. Conventional morphology and productivity studies have only compared the results of macro imaging of fungal pellets cultured in bulk with the averaged products in the culture medium. Filamentous fungi are multicellular and their expression differs between different hyphae. To truly understand the relationship between morphology and productivity, it is necessary to compare the morphology and productivity of individual hyphae. To achieve this, we developed a microfluidic system that confines hyphae to individual channels for observation and investigated the relationship between their growth, morphology, and enzyme productivity. Furthermore, using Trichoderma reesei, a potent cellulase-producing fungus, as a model, we developed a cellulase detection assay with 4-MUC substrate to detect hyphal growth and enzyme secretion in a microfluidic device in real time. Using a strain that expresses cellobiohydrolase I (CBH I) fused with AcGFP1, we compared fluorescence from the detection assay with GFP fluorescence intensity, which showed a strong correlation between the two. These results indicate that extracellular enzymes can be easily detected in the microfluidic device in real time because the production of cellulase is synchronized in T. reesei. This microfluidic system enables real-time visualization of the dynamics of hypha and enzymes during carbon source exchange and the quantitative dynamics of gene expression. This technology can be applied to many biosystems from bioenergy production to human health.

          Related collections

          Most cited references42

          • Record: found
          • Abstract: not found
          • Article: not found

          Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4

            • Record: found
            • Abstract: not found
            • Article: not found

            A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding

              • Record: found
              • Abstract: found
              • Article: not found

              Soft lithography for micro- and nanoscale patterning.

              This protocol provides an introduction to soft lithography--a collection of techniques based on printing, molding and embossing with an elastomeric stamp. Soft lithography provides access to three-dimensional and curved structures, tolerates a wide variety of materials, generates well-defined and controllable surface chemistries, and is generally compatible with biological applications. It is also low in cost, experimentally convenient and has emerged as a technology useful for a number of applications that include cell biology, microfluidics, lab-on-a-chip, microelectromechanical systems and flexible electronics/photonics. As examples, here we focus on three of the commonly used soft lithographic techniques: (i) microcontact printing of alkanethiols and proteins on gold-coated and glass substrates; (ii) replica molding for fabrication of microfluidic devices in poly(dimethyl siloxane), and of nanostructures in polyurethane or epoxy; and (iii) solvent-assisted micromolding of nanostructures in poly(methyl methacrylate).

                Author and article information

                Front Microbiol
                Front Microbiol
                Front. Microbiol.
                Frontiers in Microbiology
                Frontiers Media S.A.
                03 March 2023
                : 14
                : 1125760
                [1] 1Department of Bioengineering, Nagaoka University of Technology , Nagaoka, Japan
                [2] 2Department of Science of Technology Innovation, Nagaoka University of Technology , Nagaoka, Japan
                Author notes

                Edited by: Yang-Chun Yong, Jiangsu University, China

                Reviewed by: Lea Atanasova, University of Natural Resources and Life Sciences, Austria; Mohamed Abdelwahab Hassan, University Hospital Jena, Germany

                *Correspondence: Yosuke Shida, yshida@ 123456vos.nagaokaut.ac.jp

                This article was submitted to Microbiotechnology, a section of the journal Frontiers in Microbiology

                Copyright © 2023 Itani, Shida and Ogasawara.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                : 16 December 2022
                : 16 February 2023
                Page count
                Figures: 4, Tables: 0, Equations: 0, References: 42, Pages: 10, Words: 6976
                Original Research

                Microbiology & Virology
                filamentous fungi,hyphal growth,cellulase,microfluidic device,real-time monitoring


                Comment on this article