TY - CONF AB - Weeds pose severe threats to agricultural yields and areas used for settlement and transport. Until now, glyphosate is widely applied to reduce undesired plants worldwide [1]. Due to its negative impact on the environment, insects and humans [2-5] and likely prohibition in the EU until the end of 2023 [6], an effective as well as ecologically acceptable alternative is urgently needed. 7-deoxy-sedoheptulose (7dSh) is a novel herbicidal candidate. This sugar was isolated from culture supernatants of Synechococcus elongatus and has recently been reported to act as an inhibitor of a key enzyme of the shikimate pathway [7] analogous to glyphosate [1]. Yet the cyanobacterium S. elongatus produces very low amounts of 7dSh merely in the stationary phase of cultivation [7] and is therefore not a suitable candidate for an industrial production. Besides, chemoenzymatic synthesis of 7dSh has a low yield (20%) [7] and is not economically feasible due to high substrate costs such as 5-deoxy-D-ribose. Therefore, we aim to develop a microbial process with Streptomyces setonensis as a natural producer strain [7] for the large-scale production of 7dSh as an alternative herbicide. To evolve a scalable, well-characterized bioprocess for an unknown microbial system, firstly, parameters as media composition, phosphate and nitrogen limitation were examined using a high-throughput microbioreactor system (BioLector) which allows online-monitoring of growth. Moreover, a design of experiments (DOE) approach in shake flasks was conducted to investigate the influence of cultivation factors on each other. Results from the DOE approach suggest that elevated osmolarity through NaCl addition enables higher product titers whereas nitrogen starvation or addition has no influence. For other Streptomyces strains it is reported that the synthesis of antibiotics and other secondary metabolites is coupled to phosphate limitation [8]. Surprisingly, in contrast, 7dSh synthesis is enhanced by adding 150% more of both phosphate components than in the basic minimal medium [9] and decreased if phosphate starvation occurs. Therefore, the identification of the trigger for the 7dSh production is still ongoing, although the product titer was already increased by factor 100 from mg/L [7] to g/L. The identified beneficial culture conditions lay the foundation for the process scale-up to a stirred-tank reactor and to further improve product concentration by applying process control and defining the optimal operation regime. Moreover, organic residuals should be employed as substrates for a sustainable and cost-efficient process. REFERENCES [1] https://doi.org/10.1002/ps.1518 [2] https://doi.org/10.1038/s42003-021-02057-6 [3] https://doi.org/10.1371/journal.pbio.3001182 [4] DOI:10.3390/insects10100354 [5] DOI:10.1016/j.envpol.2020.114372 [6] https://www.bmuv.de/themen/wasser-ressourcen-abfall/boden-und-altlasten/bodenschutz-und-altlasten-worum-geht-es/faq-plan-zum-glyphosat-ausstieg [7] https://doi.org/10.1038/s41467-019-08476-8 [8] DOI: 10.1128/JB.186.16.5197-5201.2004 [9] DOI: 10.3389/fmicb.2018.02680 AU - Steurer, Xenia Ricarda AU - Jakobs-Schönwandt, Dèsirèe AU - Grünberger, Alexander AU - Patel, Anant ID - 4363 TI - Development of a bioprocess for the production of a herbicidal sugar as sustainable alternative to glyphosate ER - TY - CONF AB - Weeds pose severe threats to agricultural yields and areas used for settlement and transport. Until now, glyphosate is widely applied to reduce undesired plants worldwide [1]. Due to its negative impact on the environment, insects and humans [2-5] and likely prohibition in the EU until the end of 2023 [6], an effective as well as ecologically acceptable alternative is urgently needed. Repeated application of commercially available nontriazine herbicides caused resistant biotypes of weeds, which requires evolved agents for their control [7]. Promising alternatives to replace these harmful or inefficient products are novel herbicidal substances that target pathways in plants, that cannot be bypassed and arrest growth or cause cell death without harmful side effects on non-target organisms [8]. 7-deoxy-sedoheptulose (7dSh) is such a novel herbicidal candidate. This sugar was isolated from culture supernatants of Synechococcus elongatus and has recently been reported to act as an inhibitor of a key enzyme of the shikimate pathway [8] analogous to glyphosate [1]. Yet the cyanobacterium S. elongatus produces very low amounts of 7dSh merely in the stationary phase of cultivation [8] and is therefore not a suitable candidate for an industrial production. Besides, chemoenzymatic synthesis of 7dSh has a low yield (20%) [8] and is not economically feasible due to high substrate costs such as 5-deoxy-D-ribose. Therefore, we aim to develop a microbial process with Streptomyces setonensis as a natural producer strain [8] for the large-scale production of 7dSh as an alternative herbicide. To evolve a scalable, well-characterized bioprocess for an unknown microbial system, firstly, parameters as media composition, phosphate and nitrogen limitation were examined using a high-throughput microbioreactor system (BioLector) which allows online-monitoring of growth. Moreover, a design of experiments (DOE) approach in shake flasks was conducted to investigate the influence of cultivation factors on each other. To verify the influence of oxygen availability and gain a deeper understanding of the metabolism the oxygen transfer rate is measured with RAMOS® (Respiration Activity MOnitoring System). Results from the DOE approach suggest that elevated osmolarity through NaCl addition enables higher product titers whereas nitrogen starvation or addition has no influence. For other Streptomyces strains it is reported that the synthesis of antibiotics and other secondary metabolites is coupled to phosphate limitation [9]. Surprisingly, in contrast, 7dSh synthesis is enhanced by adding 150% more of both phosphate components than in the basic minimal medium [10] and decreased if phosphate starvation occurs. Therefore, the identification of the trigger for the 7dSh production is still ongoing, although the product titer was already increased by factor 100 from mg/L [8] to g/L. The identified beneficial culture conditions lay the foundation for the process scale-up to a stirred-tank reactor and to further improve product concentration by applying process control and defining the optimal operation regime. REFERENCES [1] Duke, S. O. and Powles, S. B. Glyphosate: a once-in-a-century herbicide. Pest. Manag. Sci. 64, 319–325 (2008). [2] Kiefer, J.S.T., Batsukh, S., Bauer, E. et al. Inhibition of a nutritional endosymbiont by glyphosate abolishes mutualistic benefit on cuticle synthesis in Oryzaephilus surinamensis. Commun Biol 4, 554 (2021). [3] Smith, D.F.Q., Camacho, E., Thakur, R. et al. Glyphosate inhibits melanization and increases susceptibility to infection in insects. PLOS Biology 19(5): e3001182 (2021). [4] Farina, W.M., Balbuena, M.S., Herbert, L.T. et al. Effects of the Herbicide Glyphosate on Honey Bee Sensory and Cognitive Abilities: Individual Impairments with Implications for the Hive. Insects 10, 354 (2019). [5] Meftaul, I.M., Venkateswarlu, K., Dharmarajan, R. et al. Controversies over human health and ecological impacts of glyphosate: Is it to be banned in modern agriculture? Environ Pollut. 263(Pt A):114372 (2020). [6] https://www.bmuv.de/themen/wasser-ressourcen-abfall/boden-und-altlasten/bodenschutz-und-altlasten- worum-geht-es/faq-plan-zum-glyphosat-ausstieg [7] LeBaron, H.M., Hill, E.R. Weeds resistant to nontriazine classes of herbicides. In: LeBaron HM, McFarland JE, Burnside OC (eds) The Triazine Herbicides. Elsevier, Chapter 11 (2008) [8] Brilisauer, K., Rapp, J., Rath, P. et al. Cyanobacterial antimetabolite 7-deoxy-sedoheptulose blocks the shikimate pathway to inhibit the growth of prototrophic organisms. Nat Commun 10, 545 (2019) [9] Martin, J.F. Phosphate control of the biosynthesis of antibiotics and other secondary metabolites is mediated by the PhoR-PhoP system: an unfinished story. J. Bacteriol. 186 (16), 5197–5201 (2004) [10] Koepff, J., Sachs, C.C., Wiechert, W. et al. Germination and Growth Analysis of Streptomyces lividans at the Single-Cell Level Under Varying Medium Compositions. Front. Microbiol. 9:2680 (2018) AU - Steurer, Xenia Ricarda AU - Jakobs-Schönwandt, Dèsirèe AU - Forchhammer, Karl AU - Grünberger, Alexander AU - Patel, Anant ID - 4362 TI - Development of a bioprocess for the production of a herbicidal sugar as sustainable alternative to glyphosate ER - TY - JOUR AB - Polyvinyl alcohol (PVA) is a biodegradable, water-soluble polymer with excellent film forming properties, commonly studied or used as tablet coating, food packaging or controlled release fertilizers. Attract-and-kill (AK) beads are sustainable, microbial alternatives to synthetic soil insecticides, whose onset of lethal effect largely depend on how fast the encapsulated entomopathogenic fungus forms virulent conidia. Therefore, the objective of this study was to develop a water-soluble coating accelerating the kill effect of AK beads by immediately releasing virulent Metarhizium brunneum CB15-III blastospores. We assessed three PVA types (PVA 4-88, 8-88, 10-98) which differed in their degree of hydrolysis or molecular weight for their ability to release viable blastospores from thin films after drying at 60–40 °C, and examined how polyethylene glycol and soy-lecithin impact the blastospore survival. Finally, we evaluated the effectiveness of coated AK beads in a bioassay against Tenebrio molitor larvae. The blastospore release rate quadrupled within the first 5 min with decreasing molecular weight and degree of hydrolysis, with PVA 4-88 releasing 79 ± 19% blastospores. Polyethylene glycol and soy-lecithin significantly increased the blastospore survival to 18–28% for all three PVA types. Coated beads exhibited a uniform, 22.4 ± 7.3 µm thin coating layer, with embedded blastospores, as confirmed by scanning electron microscopy. The blastospore coating increased the mortality rate of T. molitor larvae over uncoated AK beads, decreasing the median lethal time from 10 to 6 days. Consequently, the blastospore coating accelerated the kill effect of regular AK beads. These findings pave the way to enhanced pest control efficacy from coated systems such as beads or seeds. AU - Hermann, Katharina M. AU - Grünberger, Alexander AU - Patel, Anant ID - 3654 IS - 1 JF - AMB Express KW - formulation KW - coating KW - sustainable crop protection KW - PVA TI - Polyvinyl alcohol coating releasing fungal blastospores improves kill effect of attract-and-kill beads VL - 13 ER - TY - JOUR AB - Despite being a promising feedstock for food, feed, chemicals, and biofuels, microalgal production processes are still uneconomical due to slow growth rates, costly media, problematic downstreaming processes, and rather low cell densities. Immobilization via entrapment constitutes a promising tool to overcome these drawbacks of microalgal production and enables continuous processes with protection against shear forces and contaminations. In contrast to biopolymer gels, inorganic silica hydrogels are highly transparent and chemically, mechanically, thermally, and biologically stable. Since the first report on entrapment of living cells in silica hydrogels in 1989, efforts were made to increase the biocompatibility by omitting organic solvents during hydrolysis, removing toxic by-products, and replacing detrimental mineral acids or bases for pH adjustment. Furthermore, methods were developed to decrease the stiffness in order to enable proliferation of entrapped cells. This review aims to provide an overview of studied entrapment methods in silica hydrogels, specifically for rather sensitive microalgae. AU - Homburg, Sarah Vanessa AU - Patel, Anant ID - 1732 IS - 7 JF - Polymers TI - Silica Hydrogels as Entrapment Material for Microalgae VL - 14 ER - TY - GEN AU - Fladung, Laura AU - Homburg, Sarah Vanessa AU - Kruse, Olaf AU - Patel, Anant ID - 3165 IS - 9 T2 - Chemie Ingenieur Technik TI - Development of novel silica hydrogels with improved structure properties to support growth of entrapped microalgae VL - 94 ER - TY - CONF AU - Fladung, Laura AU - Homburg, Sarah Vanessa AU - Kruse, Olaf AU - Patel, Anant ID - 3164 IS - 9 T2 - Chemie Ingenieur Technik TI - A novel method to measure diffusion of dissolved CO2 in hydrogels VL - 94 ER - TY - GEN AU - Fladung, Laura AU - Homburg, Sarah Vanessa AU - Kruse, Olaf AU - Patel, Anant ID - 3155 TI - Development of novel silica hydrogels for the encapsulation of photosynthetic microalgae ER - TY - CONF AU - Moorlach, Benjamin Willem AU - Jakobs-Schönwandt, Désirée AU - Poranen, Minna AU - Kogel, Karl-Heinz AU - Heinlein, Manfred AU - Patel, Anant ID - 3200 TI - Kationische Nanopartikelformulierungen mit dsRNA als zielspezifisches Pflanzenschutzmittel ER - TY - GEN AU - Dietsch, Robin AU - Patel, Anant AU - Jakobs-Schönwandt, Desiree ID - 3142 TI - Beschichtete Pilzsporen und deren Verwendung als Pflanzenbehandlungsmittel ER - TY - GEN AU - Joshi, Jnanada Shrikant AU - Homburg, Sarah Vanessa AU - Kruse, Olaf AU - Patel, Anant ID - 2871 TI - Robust microalgal production processes in co-cultivation with immobilized plant growth promoting bacteria ER - TY - GEN AU - Joshi, Jnanada Shrikant AU - Homburg, Sarah Vanessa AU - Kruse, Olaf AU - Patel, Anant ID - 2864 IS - 9 SN - 0009-286X T2 - Chemie Ingenieur Technik TI - High‐resolution microscopy techniques for characterization of immobilized bacteria VL - 94 ER - TY - CONF AU - Joshi, Jnanada Shrikant AU - Homburg, Sarah Vanessa AU - Kruse, Olaf AU - Patel, Anant ID - 2862 IS - 9 SN - 0009-286X T2 - Chemie Ingenieur Technik TI - Co‐cultivation of immobilized plant growth promoting bacteria for robust microalgal production processes VL - 94 ER - TY - JOUR AU - Muskat, Linda Claire AU - Przyklenk, Michael AU - Humbert, Pascal AU - Eilenberg, Jørgen AU - Patel, Anant V. ID - 2172 IS - 5 JF - Biocontrol Science and Technology SN - 0958-3157 TI - Fermentation of the psyllid-pathogenic fungus sp. nov. inedit. (Entomophthorales: Entomophthoraceae) VL - 32 ER - TY - JOUR AU - Muskat, Linda Claire AU - Jiang, Lin AU - Brikmann, Johannes AU - Rostás, Michael AU - Patel, Anant V. ID - 2171 IS - 10 JF - Macromolecular Materials and Engineering SN - 1438-7492 TI - Development of a Self‐Adhesive Oleogel Formulation Designed for the Slow Release of Semiochemicals VL - 307 ER - TY - JOUR AB - The potential of semiochemicals for the targeted behavior manipulation of insects has been known for a long time. Their low impact on non-target organisms makes them interesting candidates for use in insect control for various applications such as agricultural and forestry pest control, stored-product protection and protection against hematophagous insects. Due to their high volatility and chemical instability against UV light and oxidation, their use often remains limited. Tailor-made formulations can protect semiochemicals from environmental factors and can improve release performance and duration triggering a desired reaction in the target insect at the place of application. This review provides an overview of recent formulation types developed for insect behavior manipulation by semiochemicals, with emphasis on formulation aspects and formulation potential for slow and controlled release. The focus was set on inventions and studies aiming to elucidate material and formulation properties that affect the semiochemical release and, with specific attention, enable targeted release manipulation. AU - Muskat, Linda Claire AU - Patel, Anant ID - 1550 JF - Entomologia Generalis SN - 0171-8177 TI - Innovations in semiochemical formulations ER - TY - CONF AB - Establishing production of a herbicidal sugar as sustainable alternative to glyphosate by a microbial chassis Steurer, X.1, Jakobs-Schönwandt, D.1, Forchhammer, K.2 and Patel, A.V.1 1University of Applied Sciences Bielefeld, Germany, 2University of Tübingen, Germany Until now, glyphosate is widely applied to reduce weeds worldwide. Due to its negative impact on the environment, insects and humans [1] and likely prohibition in the EU until the end of 2023, an effective as well as ecologically acceptable alternative is urgently needed. 7-deoxy-sedoheptulose (7dSh) might be such an alternative. This sugar is a novel herbicidal substance and was isolated from culture supernatants of Synechococcus elongatus [2]. Yet this cyanobacterium produces very low amounts of 7dSh in the stationary phase of cultivation [2] and is therefore not an adequate candidate for an industrial production. Unfortunately, chemoenzymatic synthesis of 7dSh has a low yield (20%) [2] and is not economically feasible because of the high costs of the substrate 5-deoxy-D-ribose. Therefore, we aim to provide a microbial chassis for large scale production of 7dSh. A promising option is 7dSh synthesis with Streptomyces setonensis as a natural producer strain [2]. First results indicate that production can be improved by mainly optimizing process control and culture conditions. To establish a continuous process in the bioreactor, cell immobilization will be explored. REFERENCES [1] Meftaul, I.M., Venkateswarlu, K., Dharmarajan, R. et al. Controversies over human health and ecological impacts of glyphosate: Is it to be banned in modern agriculture? Environ Pollut. 263(Pt A):114372 (2020). [2] Brilisauer, K., Rapp, J., Rath, P. et al. Cyanobacterial antimetabolite 7-deoxy-sedoheptulose blocks the shikimate pathway to inhibit the growth of prototrophic organisms. Nat Commun 10, 545 (2019). AU - Steurer, Xenia Ricarda AU - Jakobs-Schönwandt, Desiree AU - Forchhammer, Karl AU - Patel, Anant ID - 3199 IS - 9 SN - 0009-286 X T2 - Special Issue: (Bio)Process Engineering – a Key to Sustainable Development: ProcessNet and DECHEMA‐BioTechNet Jahrestagungen 2022 together with 13th ESBES Symposium TI - Establishing production of a herbicidal sugar as sustainable alternative to glyphosate by a microbial chassis VL - 94 ER - TY - JOUR AB - The control of root-feeding wireworms has become more challenging as synthetic soil insecticides have been progressively phased out due to environmental risk concerns. Innovative microbial control alternatives such as the so-called attract-and-kill strategy depend on the rapid and successful development of dried encapsulated microorganisms, which is initiated by rehydration. Casein is a functional additive that is already used in food or pharmaceutical industry due to its water binding capacity. Cross-linked forms such as formalin-casein (FC), exhibit altered network structures. To determine whether FC influences the rehydration of alginate beads in order to increase the efficacy of an attract-and-kill formulation for wireworm pest control, we incorporated either casein or FC in different alginate/starch formulations. We investigated the porous properties of alginate/ starch beads and subsequently evaluated the activities of the encapsulated entomopathogenic fungus Metarhizium brunneum and the CO2 producing yeast Saccharomyces cerevisiae. Adding caseins altered the porous structure of beads. FC decreased the bead density from (1.0197 ± 0.0008) g/mL to (1.0144 ± 0.0008) g/mL and the pore diameter by 31%. In contrast to casein, FC enhanced the water absorbency of alginate/starch beads by 40%. Furthermore, incorporating FC quadrupled the spore density on beads containing M. brunneum and S. cerevisiae, and simultaneous venting increased the spore density even by a factor of 18. Moreover, FC increased the total CO2 produced by M. brunneum and S. cerevisiae by 29%. Thus, our findings suggest that rehydration is enhanced by larger capillaries, resulting in an increased water absorption capacity. Our data further suggest that gas exchange is improved by FC. Therefore, our results indicate that FC enhances the fungal activity of both fungi M. brunneum and S. cerevisiae, presumably leading to an enhanced attract-and-kill efficacy for pest control. AU - Hermann, Katharina M. AU - Grünberger, Alexander AU - Patel, Anant ID - 2875 IS - 9 JF - World Journal of Microbiology and Biotechnology SN - 0959-3993 TI - Formalin-casein enhances water absorbency of calcium alginate beads and activity of encapsulated Metarhizium brunneum and Saccharomyces cerevisiae VL - 37 ER - TY - CONF AU - Patel, Anant AU - Jakobs-Schönwandt, Désirée AU - Moorlach, Benjamin Willem AU - Poranen, Minna AU - Kogel, Karl-Heinz AU - Heinlein, Manfred ID - 3203 TI - BioProtect -Target specific bioprotectants for sustainable crop production in a changing climate ER - TY - GEN AU - Fladung, Laura AU - Homburg, Sarah Vanessa AU - Kruse, Olaf AU - Patel, Anant ID - 3161 TI - Development of novel silica hydrogels with improved structure properties to support growth of entrapped diatoms ER - TY - CONF AU - Fladung, Laura AU - Homburg, Sarah Vanessa AU - Kruse, Olaf AU - Patel, Anant ID - 3159 TI - Development of novel silica hydrogels for the production of extracellular compounds with encapsulated diatoms ER -