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    Batch and continuous lactic acid fermentation based on a multi-substrate approach
    (Basel : MDPI AG, 2020) Olszewska-Widdrat, Agata; Alexandri, Maria; López-Gómez, José Pablo; Schneider, Roland; Venus, Joachim
    The utilisation of waste materials and industrial residues became a priority within the bioeconomy concept and the production of biobased chemicals. The aim of this study was to evaluate the feasibility to continuously produce L-lactic acid from different renewable substrates, in a multi-substrate strategy mode. Based on batch experiments observations, Bacillus coagulans A534 strain was able to continuously metabolise acid whey, sugar beet molasses, sugar bread, alfalfa press green juice and tapioca starch. Additionally, reference experiments showed its behaviour in standard medium. Continuous fermentations indicated that the highest productivity was achieved when molasses was employed with a value of 10.34 g·L−1·h−1, while the lactic acid to sugar conversion yield was 0.86 g·g−1 . This study demonstrated that LA can be efficiently produced in continuous mode regardless the substrate, which is a huge advantage in comparison to other platform chemicals. © 2020 by the authors. Licensee MDPI, Basel, Switzerland.
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    Integration of Solid State and Submerged Fermentations for the Valorization of Organic Municipal Solid Waste
    (Basel : MDPI, 2021) Martău, Gheorghe-Adrian; Unger, Peter; Schneider, Roland; Venus, Joachim; Vodnar, Dan Cristian; López-Gómez, José Pablo
    Solid state fermentation (SsF) is recognized as a suitable process for the production of enzymes using organic residues as substrates. However, only a few studies have integrated an evaluation of the feasibility of applying enzymes produced by SsF into subsequent hydrolyses followed by the production of target compounds, e.g., lactic acid (LA), through submerged-liquid fermentations (SmF). In this study, wheat bran (WB) was used as the substrate for the production of enzymes via SsF by Aspergillus awamori DSM No. 63272. Following optimization, cellulase and glucoamylase activities were 73.63 ± 5.47 FPU/gds and 107.10 ± 2.63 U/gdb after 7 days and 5 days of fermentation, respectively. Enzymes were then used for the hydrolysis of the organic fraction of municipal solid waste (OFMSW). During hydrolysis, glucose increased considerably with a final value of 19.77 ± 1.56 g/L. Subsequently, hydrolysates were fermented in SmF by Bacillus coagulans A166 increasing the LA concentration by 15.59 g/L. The data reported in this study provides an example of how SsF and SmF technologies can be combined for the valorization of WB and OFMSW.
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    Limited life cycle andcost assessment for the bioconversion of lignin‐derived aromatics into adipic acid
    (New York, NY [u.a.] : Wiley, 2020) van Duuren, Jozef B.J.H.; de Wild, Paul J.; Starck, Sören; Bradtmöller, Christian; Selzer, Mirjam; Mehlmann, Kerstin; Schneider, Roland; Kohlstedt, Michael; Poblete‐Castr, Ignacio; Stolzenberger, Jessica; Barton, Nadja; Fritz, Michel; Scholl, Stephan; Venus, Joachim; Wittmann, Christoph
    Lignin is an abundant and heterogeneous waste byproduct of the cellulosic industry, which has the potential of being transformed into valuable biochemicals via microbial fermentation. In this study, we applied a fast-pyrolysis process using softwood lignin resulting in a two-phase bio-oil containing monomeric and oligomeric aromatics without syringol. We demonstrated that an additional hydrodeoxygenation step within the process leads to an enhanced thermochemical conversion of guaiacol into catechol and phenol. After steam bath distillation, Pseudomonas putida KT2440-BN6 achieved a percent yield of cis, cis-muconic acid of up to 95 mol% from catechol derived from the aqueous phase. We next established a downstream process for purifying cis, cis-muconic acid (39.9 g/L) produced in a 42.5 L fermenter using glucose and benzoate as carbon substrates. On the basis of the obtained values for each unit operation of the empirical processes, we next performed a limited life cycle and cost analysis of an integrated biotechnological and chemical process for producing adipic acid and then compared it with the conventional petrochemical route. The simulated scenarios estimate that by attaining a mixture of catechol, phenol, cresol, and guaiacol (1:0.34:0.18:0, mol ratio), a titer of 62.5 (g/L) cis, cis-muconic acid in the bioreactor, and a controlled cooling of pyrolysis gases to concentrate monomeric aromatics in the aqueous phase, the bio-based route results in a reduction of CO2-eq emission by 58% and energy demand by 23% with a contribution margin for the aqueous phase of up to 88.05 euro/ton. We conclude that the bio-based production of adipic acid from softwood lignins brings environmental benefits over the petrochemical procedure and is cost-effective at an industrial scale. Further research is essential to achieve the proposed cis, cis-muconic acid yield from true lignin-derived aromatics using whole-cell biocatalysts. © 2020 Wiley Periodicals, Inc.
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    Pilot Scale for Production and Purification of Lactic Acid from Ceratonia siliqua L. (Carob) Bagasse
    (Basel : MDPI, 2022) Azaizeh, Hassan; Abu Tayeh, Hiba Nazmi; Schneider, Roland; Venus, Joachim
    The bioconversion of lignocellulose and organic waste bagasse to lactic acid (LA) is an important alternative process requiring valorization as a potentially viable method in the production of pure LA, to be utilized for various purposes. Carob (Ceratonia siliqua L.) biomass was used for the production of LA, using a thermophilic Bacillus coagulans isolate, cultivated in a batch pilot scale of 35 L fermenters without yeast extract supplementation, and operated for 50 h. During the fermentation process, most of the degradable sugar was consumed within 35 h and resulted in the production of 46.9 g/L LA, with a calculated LA yield of 0.72 g/g sugars and productivity at the log phase of 1.69 g/L/h. The use of LA for different industrial applications requires high purity; therefore, a downstream process (DSP) consisting of different purification stages was used, enabling us to reach up to 99.9% (w/w) product purity, which indicates that the process was very effective. The overall almost pure L-LA yield of the DSP was 56%, which indicates that a considerable amount of LA (46%) was lost during the different DSP stages. This is the first study in which carob biomass bagasse has been tested on a pilot scale for LA production, showing the industrial feasibility of the fermentation process.
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    Assessing the organic fraction of municipal solid wastes for the production of lactic acid
    (Amsterdam [u.a.] : Elsevier, 2019) López-Gómez, J. Pablo; Latorre-Sánchez, Marcos; Unger, Peter; Schneider, Roland; Coll Lozano, Caterina; Venus, Joachim
    With an estimated yearly production of about 140 Mt in the EU, conventionally, the organic fraction of municipal solid wastes (OFMSW) has been disposed in landfills with negative environmental effects. Nonetheless, the chemical composition of this residue make it a substrate with great bioconversion potential. In this study, OFMSW from Spanish municipal treatment plants, were evaluated for the production of LA. Samples were identified according to the sorting mechanisms employed for their collection in: (A) separately collected, (B) non-separately collected and (C) separately collected+paper/cardboard. Enzymatic hydrolysis was used to produce hydrolysates A, B and C accordingly. Hydrolysate A showed the highest total sugars and glucose content with values of 70 and 55 g·L−1, respectively. Following the characterisation, a screening showed that growth of B. coagulans was possible in all three hydrolysates. Furthermore, lab scale fermentations showed that LA final concentrations could reach around 60 g·L−1, with yields from total sugars of above 0.60 g·g−1. A technical scale fermentation of the hydrolysate A resulted in a final LA concentration of 60.7 g·L−1, a yield of 0.71 g·g−1 with a productivity of 2.68 g·L−1·h−1. Overall, it was estimated that 0.23 g of LA could be produced from one g of dry OFMSW.
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    Valorising agro-industrial wastes within the circular bioeconomy concept: The case of defatted rice bran with emphasis on bioconversion strategies
    (Basel : MDPI AG, 2020) Alexandri, Maria; López-Gómez, José Pablo; Olszewska-Widdrat, Agata; Venus, Joachim
    The numerous environmental problems caused by the extensive use of fossil resources have led to the formation of the circular bioeconomy concept. Renewable resources will constitute the cornerstone of this new, sustainable model, with biomass presenting a huge potential for the production of fuels and chemicals. In this context, waste and by-product streams from the food industry will be treated not as "wastes" but as resources. Rice production generates various by-product streams which currently are highly unexploited, leading to environmental problems especially in the countries that are the main producers. The main by-product streams include the straw, the husks, and the rice bran. Among these streams, rice bran finds applications in the food industry and cosmetics, mainly due to its high oil content. The high demand for rice bran oil generates huge amounts of defatted rice bran (DRB), the main by-product of the oil extraction process. The sustainable utilisation of this by-product has been a topic of research, either as a food additive or via its bioconversion into value-added products and chemicals. This review describes all the processes involved in the efficient bioconversion of DRB into biotechnological products. The detailed description of the production process, yields and productivities, as well as strains used for the production of bioethanol, lactic acid and biobutanol, among others, are discussed. © 2020 by the authors. Licensee MDPI, Basel, Switzerland.
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    Towards efficient production of highly optically pure d-lactic acid from lignocellulosic hydrolysates using newly isolated lactic acid bacteria
    (New York, NY [u.a.] : Elsevier, 2022) Alexandri, Maria; Hübner, Dennis; Schneider, Roland; Fröhling, Antje; Venus, Joachim
    This study presents the production of D-lactic acid with high enantiomeric purity using lignocellulosic hydrolysates from newly isolated lactic acid bacterial (LAB) strains. Six strains, 4 heterofermentative and 2 homofermentative, were investigated for their ability to grow and produce lactic acid on sugar beet pulp (SBP) hydrolysates, containing a mixture of hexose and pentose sugars. Among the strains tested, three were isolates designated as A250, A257 and A15, all of which belonged to the genus Leuconostoc. Only strain A250 could be reliably identified as Leuconostoc pseudomesenteroides based on cluster analysis of Maldi-ToF spectra. All strains produced D-lactic acid in the presence of SBP hydrolysates, but with varying optical purities. The homofermentative strains achieved higher D-lactic acid optical purities, but without assimilating the pentose sugars. Co-cultivation of the homofermentative strain Lactobacillus coryniformis subsp. torquens DSM 20005 together with the heterofermentative isolate A250 led to the production of 21.7 g/L D-lactic acid with 99.3 % optical purity. This strategy enabled the complete sugar utilization of the substrate. Nanofiltration of the SBP hydrolysate enhanced the enantiomeric purity of the D-lactic acid produced from the isolates A250 and A15 by about 5 %. The highest D-lactic acid concentration (40 g/L) was achieved in fed-batch cultures of A250 isolate with nanofiltered SBP, where optical purity was 99.4 %. The results of this study underline the feasibility of a novel isolate as an efficient D-lactic acid producer using lignocellulosic hydrolysates.
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    Production of lactic acid from pasta wastes using a biorefinery approach
    (London : BioMed Central, 2022) Marzo-Gago, Cristina; Venus, Joachim; López-Gómez, José Pablo
    A total of 398 kt of pasta waste (PW), generated during the production process of pasta, were produced in 2021. Due to its chemical composition and practically zero cost, PW has already been studied as a raw material for the production of lactic acid (LA) through fermentations. The main objective of this article was to improve the economic viability of the process by replacing commercial enzymes, necessary for starch hydrolysis in PW, with raw enzymes also produced from wastes. Enzyme synthesis was achieved through solid-state fermentation (SsF) of wheat bran by Aspergillus awamori or Aspergillus oryzae at various moisture contents. The maximum amylase activity (52 U/g dry solid) was achieved after 2 days of fermentation with A. awamori at 60% of moisture content. After that, the enzymes were used to hydrolyse PW, reaching 76 g/L of total sugars, 65 g/L of glucose and a yield of 0.72 gglu/gds with the enzymes produced by A. awamori. Subsequently, the hydrolysate was fermented into LA using Bacillus coagulans A559, yielding 52 g/L and 49 g/L with and without yeast extract, respectively. Remarkably, compared to the process with commercial enzymes, a higher LA yield was reached when enzymes produced by SsF were added (0.80 gLA/gglu). Furthermore, the productivities between the two processes were similar (around 3.9 g/L/h) which highlights that yeast extract is not necessary when using enzymes produced by SsF.
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    Investigation of spiral-wound membrane modules for the cross-flow nanofiltration of fermentation broth obtained from a pilot plant fermentation reactor for the continuous production of lactic acid
    (Berlin ; Heidelberg [u.a.] : SpringerOpen, 2017-1-4) Laube, Hendrik; Schneider, Roland; Venus, Joachim
    Background: The separation performance of seven polymer membranes for the nanofiltration of sodium lactate in fermentation broth was investigated. Each module was introduced into the test stand, and the system curve was obtained by recording the permeate flow velocity at different pump head levels. Performance benchmarks were good permeate quality, as determined by high permeate flow velocity, high sodium lactic concentration, low ion impurity concentration, and low organic impurity concentration. Market research has shown that three companies, DOW (TW30, SW30, NF45), General Electric (DK73, DL73), and Microdyn-Nadir (NP30), distributed spiral-wound membrane modules for cross-flow filtration in a 2.5 by 40-in. module size, suitable for operation in the filtration test stand. Results: The measured permeate flow velocity was found to vary widely between the membranes. At a pump head of 250 m, DK73, NP30, and DL73 generated more than 200, 300, and 400% higher permeate flow velocities, respectively, than TW30 and NF45. A key benchmark, lactate rejection, was also highly dependent upon membrane type. The NP30, NF45, and TW30 membranes showed a decrease in lactate permeate flow velocity of 117, 83, and 348% starting at 205, 250, and 300 m, respectively. Conclusions: The DL73 had the overall best performance according to the measured fermentation broth and lactic acid permeability. The presented method for the graphical analysis of the membrane performance proofed to be a useful tool for the filtration engineer. © 2017, The Author(s).
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    Potential Role of Sequential Solid-State and Submerged-Liquid Fermentations in a Circular Bioeconomy
    (Basel : MDPI, 2021) López-Gómez, José Pablo; Venus, Joachim
    An efficient processing of organic solid residues will be pivotal in the development of the circular bioeconomy. Due to their composition, such residues comprise a great biochemical conversion potential through fermentations. Generally, the carbohydrates and proteins present in the organic wastes cannot be directly metabolized by microorganisms. Thus, before fermentation, enzymes are used in a hydrolysis step to release digestible sugars and nitrogen. Although enzymes can be efficiently produced from organic solid residues in solid-state fermentations (SsF), challenges in the development and scale-up of SsF technologies, especially bioreactors, have hindered a wider application of such systems. Therefore, most of the commercial enzymes are produced in submerged-liquid fermentations (SmF) from expensive simple sugars. Instead of independently evaluating SsF and SmF, the review covers the option of combining them in a sequential process in which, enzymes are firstly produced in SsF and then used for hydrolysis, yielding a suitable medium for SmF. The article reviews experimental work that has demonstrated the feasibility of the process and underlines the benefits that such combination has. Finally, a discussion is included which highlights that, unlike typically perceived, SsF should not be considered a counterpart of SmF but, in contrast, the main advantages of each type of fermentation are accentuated in a synergistic sequential SsF-SmF.