Filamentous fungi are widely used in the biotechnology industry for the production of industrial enzymes. Thus, considerable work has been done with the purpose of characterizing these processes. The ultimate goal of these efforts is to be able to control and predict fermentation performance on the basis of "standardized" measurements in terms of morphology, rheology, viscosity, mass transfer and productivity. However, because the variables are connected or dependent on each other, this task is not trivial. The aim of this review article is to gather available information in order to explain the interconnectivity between the different variables in submerged fermentations. An additional factor which makes the characterization of a fermentation broth even more challenging is that the data obtained are also dependent on the way they have been collected-meaning which technologies or probes have been used, and on the way the data is interpreted-i.e. which models were applied. The main filamentous fungi used in industrial fermentation are introduced, ranging from Trichoderma reesei to Aspergillus species. Due to the fact that secondary metabolites, like antibiotics, are not to be considered bulk products, organisms like e.g. Penicillium chrysogenum are just briefly touched upon for the description of some characterization techniques. The potential for development of different morphological phenotypes is discussed as well, also in view of what this could mean to productivity and-equally important-the collection of the data. An overview of the state of the art techniques for morphology characterization is provided, discussing methods that finally can be employed as the computational power has grown sufficiently in the recent years. Image analysis (IA) clearly benefits most but it also means that methods like near infrared measurement (NIR), capacitance and on-line viscosity now provide potential alternatives as powerful tools for characterizing morphology. These measuring techniques, and to some extent their combination, allow obtaining the data necessary for supporting the creation of mathematical models describing the fermentation process. An important part of this article will indeed focus on describing the different models, and on discussing their importance to fermentations of filamentous fungi in general. The main conclusion is that it has not yet been attempted to develop an overarching model that spans across strains and scales, as most studies indeed conclude that their respective results might be strain specific and not necessarily valid across scales.
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.
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