A fast and simple method to estimate relative, hyphal tensile‐strength of filamentous fungi used to assess the effect of autophagy
Loading...
Links to Files
Author/Creator ORCID
Date
2017-11-14
Type of Work
Department
Program
Citation of Original Publication
Daniela Quintanilla, Cynthia Chelius, Sirasa Iambamrung, Sidney Nelson, Donnel Thomas, Krist V. Gernaey, Mark R. Marten, A fast and simple method to estimate relative, hyphal tensile‐strength of filamentous fungi used to assess the effect of autophagy, Biotechnology and Bioengineering, Volume 115, Issue 3 , https://doi.org/10.1002/bit.26490
Rights
This item is likely protected under Title 17 of the U.S. Copyright Law. Unless on a Creative Commons license, for uses protected by Copyright Law, contact the copyright holder or the author.
This is the peer reviewed version of the following article: Daniela Quintanilla, Cynthia Chelius, Sirasa Iambamrung, Sidney Nelson, Donnel Thomas, Krist V. Gernaey, Mark R. Marten, A fast and simple method to estimate relative, hyphal tensile‐strength of filamentous fungi used to assess the effect of autophagy, Biotechnology and Bioengineering, Volume 115, Issue 3 , https://doi.org/10.1002/bit.26490, which has been published in final form at https://doi.org/10.1002/bit.26490 . This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.
This is the peer reviewed version of the following article: Daniela Quintanilla, Cynthia Chelius, Sirasa Iambamrung, Sidney Nelson, Donnel Thomas, Krist V. Gernaey, Mark R. Marten, A fast and simple method to estimate relative, hyphal tensile‐strength of filamentous fungi used to assess the effect of autophagy, Biotechnology and Bioengineering, Volume 115, Issue 3 , https://doi.org/10.1002/bit.26490, which has been published in final form at https://doi.org/10.1002/bit.26490 . This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.
Abstract
Fungal hyphal strength is an important phenotype which can have a profound impact on bioprocess behavior. Until now, there is not an efficient method which allows its characterization. Currently available methods are very time consuming, thus, compromising their applicability in strain selection and process development. To overcome this issue, a method for fast and easy, statistically verified quantification of relative hyphal tensile strength was developed. It involves off‐line fragmentation in a high shear mixer followed by quantification of fragment size using laser diffraction. Particle size distribution (PSD) is determined, with analysis time on the order of minutes. Plots of PSD 90th percentile versus time allow estimation of the specific fragmentation rate. This novel method is demonstrated by estimating relative hyphal strength during growth in control conditions and rapamycin‐induced autophagy for Aspergillus nidulans (parental strain) and a mutant strain (ΔAnatg8) lacking an important autophagy gene. Both strains were grown in shake flasks and relative hyphal tensile strength was compared. The mutant strain grown in control conditions appears to be weaker than the parental strain, suggesting that Anatg8 may play a role in other processes involving cell wall biosynthesis. Furthermore, rapamycin‐induced autophagy resulted in apparently weaker cells even for the mutant strain. These findings confirm the utility of the developed method in strain selection and process development.