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Trichoderma atroviride IMI 206040 synthesizes the coconut lactone 6-pentyl-α-pyrone (6-PAP) de novo and Aspergillus niger DSM 821 produces the rose-like flavour compound 2-phenylethanol (2-PE) from the precursor l-phenylalanine. Here, microparticles of different chemical composition and nominal particle diameter in the range 5-250 µm were added to shake-flask cultures of both fungi to investigate the particles' effect on product formation. Maximum 2-PE concentration increased by a factor of 1.3 to 1430 mg/l with the addition of 2% w/v talc (40 µm diameter). Maximum 6-PAP concentration increased by a factor of 2 to 40 mg/l with the addition of 2% w/v iron (II, III) oxide. The influence of ions leaching out of the particles was investigated by cultivating the fungi in leached particle medium. For the first time, the positive effect of the microparticle-enhanced cultivation (MPEC) technique on the microbial production of volatile metabolites, here flavour compounds from submerged fungal cultures, is demonstrated. The effect is strain- and particle-specific.

One of the most sensitive process characteristics in the cultivation of filamentous biological systems is their complex morphology. In submerged cultures, the observed macroscopic morphology of filamentous microorganisms varies from freely dispersed mycelium to dense spherical pellets consisting of a more or less dense, branched and partially intertwined network of hyphae. Recently, the freely dispersed mycelium form has been in high demand for submerged cultivation because this morphology enhances the growth and production of several valuable products. A distinct filamentous morphology and productivity are influenced by the environment and can be controlled by inoculum concentration, spore viability, pH value, cultivation temperature, dissolved oxygen concentration, medium composition, mechanical stress or process mode as well as through the addition of inorganic salts or microparticles, which provides the opportunity to tailor a filamentous morphology. The suitable morphology for a given bioprocess varies depending on the desired product. Therefore, the advantages and disadvantages of each morphological type should be carefully evaluated for every biological system. Because of the high industrial relevance of filamentous microorganisms, research in previous years has aimed at the development of tools and techniques to characterise their growth and obtain quantitative estimates of their morphological properties. The focus of this review is on current advances in the characterisation and control of filamentous morphology with a separation of eukaryotic and prokaryotic systems. Furthermore, recent strategies to tailor the morphology through classical biochemical process parameters, morphology and genetic engineering to optimise the productivity of these filamentous systems are discussed.

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Here we describe a PCR-based analysis system that allows the simple simultaneous assessment of murine interferons (IFN)-alpha and IFN-beta induction in a single reaction. In this analysis, the so-called early IFN-alpha4 can be distinguished from the so-called late IFN-nonalpha4 by employing a primer mixture that amplifies a part of the IFN-alpha genes in which IFN-alpha4 exhibits a deletion of 15 nucleotides compared to IFN-nonalpha4. By including a final denaturation and a slow cooling step at the end of the PCR procedure, hybrid formation was avoided that regularly occurred when standard protocols were used. Separation of the amplification products on 4.5% agarose gels allowed the comparative assessment of the classical type I IFNs. Using this analysis system, we could show that in immortalized adult fibroblasts, IFN-beta is induced first and the two types of IFN-alpha are induced later and simultaneously. When similar fibroblasts derived from mice that lack IFN-beta were tested, the IFN response was delayed. However, now IFN-alpha4 appeared first and apparently induced the cascade of IFN-nonalpha4. This confirms the role of IFN-beta as master regulator of the normal IFN response.

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