IP4 Science Talks and AG Ecothermodynamics/Biocalorimetry

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Karim Fahmy

Prof. Karim Fahmy

Helmholtz-Zentrum Dresden Rossendorf (HZDR)

Institute of Resource Ecology

26.08.24, 2:30 pm in KUBUS 1CD

 

Abstract

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IP4 Science Talks and SOMA

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Jens Appel

Dr. Jens Appel

Universität Kassel

Bioenergetics in Photoautotrophs

15.05.24, 2:30 pm in Building 4.0, Room 101

 

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IP4 Science Talks and AG Biophotovoltaics

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Pablo Nickel

Prof. Pablo Nikel

Systems Environmental Biology
DTU Biosustain, Denmark

06.05.24, 09:30 am in KUBUS 1CD

 

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IP4 Science Talks and MIBITECH

'Electroactive biofilms and their applications for resource recovery'

Annemiek ter Heijnen

Prof. Annemiek ter Heijne

Environmental Technology
Wageningen University & Research

03.04.24, 2:30 pm in KUBUS 1A

 

Abstract

Electro-active microorganisms can play a role in many different conversions, by exchanging chemical energy with electrical energy (and vice versa) in so-called Microbial Electrochemical Systems. In this presentation, I will explain how microorganisms and electrodes can interact to facilitate different types of conversions, with applications in resource recovery and removal of nutrients. I will focus on different applications: how anaerobic ammonium oxidation at bioanodes can be enhanced by introducing oxygen, how we can influence storage processes in electro-active biofilms on anodes, and how we can use microorganisms at the cathode to produce methane from CO2. I will discuss the main limitations and routes for further improvement of rate and efficiency.


Short CV

Annemiek ter Heijne is professor in (Bio)Recovery Technology for Circular Economy at the Environmental Technology group of Wageningen University & Research (The Netherlands). She obtained her PhD degree on Microbial Fuel Cells and has continued her career in Wageningen working on different applications that involve the interaction between microorganisms and electrodes. She has been board member and president of ISMET, the International Society of Microbial Electrochemistry & Technology, and her teaching focuses on Renewable Energy Technologies and Microbial Electrochemistry. In her free time, she loves to spend time with her family, she likes spending time outdoor (hiking and running), and cooking (vegan) food.



IP4 Science Talks

'Tuning multi-enzyme catalysed processes – modular cascade set-up, activity regulation, and integration with microbial and chemical transformations'

Prof Doerthe Rother

Prof. Dörte Rother

Systems Biotechnology, AG Synthetic Enzyme Cascades
Forschungszentrum Jülich

28.02.24, 2:30 pm in KUBUS 1CD

 

Abstract

There is an urgent need for the development of greener syntheses procedures if we want to maintain an environment worth living in and keep a high standard in material comfort (or reach a higher one in developing
countries). The establishment of more biocatalytic steps in chemical syntheses is one possible solution, as enzymes and whole cells offer sustainable advantages, such as biodegradability, intoxicity, high selectivity, and many more. As a myriad of enzymatic reactions exist for almost any product, their potential is immense. Great scientific achievements and new techniques have enabled the design of economically and ecologically feasible one-step and multi-step enzyme catalysed reactions. This presentation will highlight the advantages of multi enzyme catalysed processes especially with respect to atom- and step efficiency, selectivity and modularity. However, with new opportunities, also new challenges arise. Two new possibilities, including our efforts to circumvent the associated disadvantages, will be subsequently presented: (A) avoiding cross-reactivity in (self-) regulated one-enzyme cascades and (B) the potential of using renewable resources as starting materials when microbial cell factories, enzymes and chemical catalysts are combined effectively. (A) The more enzyme steps are combined, the higher the risk of undesired cross-reactivity. Separation in space or on time can solve this issue. In the ‘LightCas’-project we investigate the possibilities to avoid cross-reactivity in one-pot systems by separation of reaction steps in time. With the help of sequential enzyme addition and on-demand light-induced enzyme inactivation 1 , a tight control of each biocatalytic step in a one-pot cascade is possible. This results in high product purity. Due to online analytics and automation, our ultimate goal is in close reach: setting up a one-pot multi-step light-controlled enzyme reactor yielding tetrahydroisoquinolines2 from cheap substrates with high selectivity and concentration in a technically self-controlled manner. (B) The combination of suitable chemical and biological catalysts with their intrinsic advantages holds the potential to develop processes that are truly superior to current production methods in terms of efficiency and sustainability. In such hybrid processes, e.g. microorganisms can use substrates from human waste streams from agriculture and the food industry to provide simple chemical building blocks. Then, enzymes are used to diversify these compounds in vitro, enabling the construction of product platforms of valuable fine chemicals, complex (chiral) building blocks and active pharmaceutical ingredients. In addition, chemical transformations can complement and diversify this product portfolio. This approach will be shown on the examples of (i) a hybrid process for the bio-based production of chiral amino alcohols from xylose and glucose 3,4 (Figure 1) and (ii) the adaptive use of renewable raw materials and the integration of CO2 in the chemo-enzymatic production of chiral diols and dioxolanes. Challenges, like the identification of suitable reaction conditions (including unconventional media) for each individual catalysts and the need of integrating downstream processing 6 to make these hybrid processes truly advantageous over other strategies will be discussed.

Figure 1. Enzymatic cascade towards metaraminol starting from renewables.

Figure 1. Enzymatic cascade towards metaraminol starting from renewables. The cascade comprises of a carboxylic acid reduction (1), a carboligation (2) and a transamination (3) step. The aliphatic precursors pyruvate and L -alanine can be produced microbially from second generation feedstocks (D-glucose, D-xylose) in suitable amounts. In situ product removal (ISPR) is used to shift the reaction equilibrium of the transamination step.


Short CV

Dörte Rother holds a diploma degree in biology from RWTH Aachen University. She conducted her PhD at the Heinrich-Heine University Düsseldorf (with research stays at Karolinska Institute Stockholm, University of Freiburg and University of Stuttgart). In 2008 she received a DFG-postdoc scholarship to work at Aachen University and became in 2009 postdoctoral student at the IBG-1: Biotechnology at Research Center Jülich. In 2012 she started her own Helmholtz-young investigators group in Jülich, additionally funded by a ERC-starting grant in 2017. In 2018 Dörte Rother was appointed as full professor for `synthetic enzyme cascades’ at Aachen University and leads the group `biocatalysis’ at Research Center Jülich. She has three children. She received the DECHEMA-Prize 2018 and the Biotrans Junior Award 2019 for her outstanding research activities in multi-step biocatalysis and sustainable process development.



IP4 Science Talks

'Closing cycles in the plastics bioeconomy through Pseudomonas biotechnology'

Prof Nick Wierckx

Prof. Nick Wiercx

Microbial Catalysis
Forschungszentrum Jülich

31.01.24, 2:30 pm in KUBUS, 1A

 

Abstract

Over 400 million tons of plastic are produced worldwide every year. These materials make up a cornerstone of our modern way of life, but the environmental impact of primarily fossil-based plastics has been broadly discussed. For the majority of plastics there are currently no viable end-of-life options except incineration, and the reality is that we are facing a major crisis of plastic pollution with widespread impacts on the environment and human well-being. There is thus an urgent need for new recycling technologies that can cope with the ever-increasing complexity of plastic materials.
In this context, we see great potential in the bio-upcycling of plastic waste, especially when combined with the development of new polymers and composites tailored for end-of-life functionalities such as biodegradability. Such polymers often contain biosimilar C-O and C-N heteroatom linkages, including polyesters, polyamides, and polyurethanes (1). Upon enzymatic or chemical hydrolysis of such polymers, a mixture of monomers is released containing alcohol, acid, or amine groups. Depending on the starting substrate the separation of these mixtures into their individual constituents may not be economical. In this case, we propose to use these plastic hydrolysates as carbon source for biotechnology (2). To enable this, we have developed Pseudomonas into a plastic monomer-metabolizing biotechnological workhorse, capable of metabolizing a wide range of aliphatic diols, dicarboxylates, and amines. Currently our main focus is to develop these monomer-metabolizing strains to not just grow on plastic monomers, but to convert plastic hydrolysates into value-added chemicals and biopolymers.
Besides addressing the end-of-life of plastics, it is also important to address their beginning. Currently plastics are primarily fossil-based which ultimately causes climate change, petrochemical pollution, and a dependence on politically unstable countries with poor humanitarian records. We therefore also develop Pseudomonas catalysts for the bio-based production of chemicals, including plastic building blocks. Both aspects will be discussed in the context of transitioning plastics into the circular bioeconomy.

(1) Ellis et al. (2021) Chemical and biological catalysis for plastics recycling and upcycling. Nature Catalysis 4:539-556
(2) Tiso et al. (2022) The metabolic potential of plastics as biotechnological carbon sources – Review and targets for the future. Metabolic Engineering 71:77-98


IP4 Science Talks and SOMA

'Cyanobacterial growth and the productivity of phototrophic cultures: insights from computational models'

Ralf Steuer

Dr. Ralf Steuer

Institute for Theoretical Biology
Humboldt-University Berlin

29.11.23, 3:00 pm in Building 4.0, Room 101

 

Abstract

The evolution of oxygenic photosynthesis in the ancestors of modern-day cyanobacteria gave rise to perhaps the most important biological process within our biosphere: the assimilation of atmospheric CO2 into organic biomass and the release of molecular oxygen as a byproduct. While properties of cyanobacterial growth in axenic cultures are increasingly understood, there are still many open questions regarding the limits of cyanobacterial growth. In particular, the economic viability of cyanobacterial biotechnology is still constrained by low growth rates and low areal productivities.

In the talk, I will discuss computational models of cyanobacterial metabolism and growth, from genome-scale reconstructions to coarse-grained kinetic models of growth. A particular focus will be optimality and resource allocation. That is, how do microbial and, in particular, cyanobacterial cells allocate their finite cellular resources, such as a finite proteome or a finite cytosolic space, to ensure maximal evolutionary success in a given environment. A second theme will concern the implications of resource allocation for cyanobacterial productivity. It is shown that high maximal growth rates are not a sufficient or necessary property for high phototrophic productivity. Finally, I will discuss how the perspective from (optimal) resource allocation can help us to understand the evolution of interactions between phototrophic and heterotrophic microbes as part of microbial ecosystems.


Short CV

Ralf Steuer studied Physics in Munich and Berlin. Previous works include time-series analysis, information theory, analysis of metabolomic data, and computational model of cellular metabolism. Within the past 10 years a particular focus has been on computational descriptions of cyanobacterial metabolism and growth. The research group published genome-scale metabolic reconstructions of several cyanobacterial strains and pioneered the use of large-scale models of time-dependent resource-allocation in phototrophic microorganisms.


References

[1] Bruggeman, FJ, Teusink, B, Steuer, R (2023). Trade-offs between the instantaneous growth rate and long-term fitness: Consequences for microbial physiology and predictive computational models. BioEssays, 45, e2300015.
[2] Faizi M, Steuer, R (2019) Optimal proteome allocation strategies for phototrophic growth in a light-limited chemostat. Microb Cell Fact 18, 165
[3] Zavřel T, Faizi M, et al. (2019) Quantitative insights into the cyanobacterial cell economy. eLife 8:e42508


Lecture of AGs Plant Biogeochemistry and Ecothermodynamics/Biocalorimetry

'Cadmium in cacao crop: Using bioremediation as lead strategy in Colombia'

Daniel Bravo

Dr. Daniel Bravo

AGROSAVIA, Colombia

14.09.23, 9:00 am in Building 1.0, Room 255 and online via Zoom.

 

Cacao, dear to many European chocolate consumers, often contains the toxic heavy metal cadmium. In 2019, the European Commission regulated the amount of permissible cadmium in cacao products, putting immense pressure on the South American cacao industry. Bioremediation of cacao farm soil is necessary, but also merits in-depth research. However, only a few studies approach the bioremediation of cacao soils systematically, i.e. by limiting cadmium bioavailability in soils. This talk will explore two possible ways of cadmium bioremediation in cacao soils. The first one, a field-related technique based on thermodynamics, such as the isothermal microcalorimetry, to select a group of cadmium-tolerant bacteria. The second one, the application of bacterial inocula to increase the immobilization of cadmium under field conditions.


IP4 Science Talks

'Chemoproteomics to Identify Metabolite Regulation of Photosynthesis and Carbon Fixations'

Prof. Paul Hudson

Prof. Paul Hudson

School of Biotechnology
KTH Royal institute of Technology Stockholm, Sweden

13.09.23, 2 pm in KUBUS hall 1A

 

Metabolic engineering of microbes to produce compounds of interest often involves dramatic alteration of metabolite pools. An often-overlooked consequence is that accumulating metabolites may negatively affect enzyme activity through competitive or allosteric inhibition. Additionally, post-translational regulation of enzyme activity can complicate metabolic engineering, as pathway flux becomes less sensitive to artificial enzyme overexpression. Here I will describe our adaptation of two chemoproteomics methods to identify metabolite-protein interactions in the proteomes of cyanobacteria and chloroplasts. These techniques rely on changes in thermal stability or of proteolytic digestion susceptibility in the presence of an effector metabolite, and together allow us to map metabolite-protein binding surfaces. In an initial effort, we found widespread interaction of tested metabolites with enzymes in central carbon metabolism, but that only a fraction of these interactions affect catalysis. For example, the Calvin cycle intermediate glyceraldehyde phosphate (GAP) stimulated activity of the Calvin cycle enzyme F/SBPase in reducing conditions, representing a feed-forward activation of the cycle. In oxidizing conditions however, GAP inhibited the enzyme by promoting aggregation. I will also describe efforts in mutagenizing enzymes to reduce sensitivity to metabolite regulation, by high-throughput screening of the effects of enzyme mutation on both cell growth and product synthesis rates.

Paul Hudson is associate professor in the School of Biotechnology at KTH Royal Institute of Technology in Stockholm, Sweden. He studied chemical engineering at NC State and UC Berkeley, and did post-doctoral training at KTH on bacterial surface display (2012-2014). His research as a group leader is on metabolic engineering of CO2-fixing bacteria, with focus on Calvin cycle bacteria such as cyanobacteria. The research uses systems biology methods such as proteomics to study CO2 fixation metabolism, and to guide engineering strategies for enhancing CO2 uptake and conversion. His group also develops synthetic biology tools in these strains.



IP4 Science Talks

'Climate extremes alter controls and pathways of soil carbon loss from alpine floodplains'

Marco Keiluweit

Prof. Marco Keiluweit

Soil & Microbial Biogeochemistry

Institute of Earth Surface Dynamics at the University of Lausanne, Switzerland

28.06.23, 1 pm in KUBUS hall 1A

 

Floodplains within mountainous catchments are dynamic reservoirs of carbon and nutrients, and experience seasonal flooding due to snowmelt and drainage. Climate change is rapidly altering snowpack and melt dynamics, resulting in greater variability in flooding and droughts. The variable hydrology drives spatial and temporal redox gradients within floodplain soils, with largely unknown consequences for carbon storage in and export from such ecosystems. In this presentation, I will report on a high-resolution monitoring campaign across extremely low and high river discharge years, foreshadowing climate change predictions, in a headwater catchment in the Rocky Mountains (Colorado, US). Combining in-field biogeochemical measurements with molecular and geochemical measurements, I will show that such extreme hydrological extremes undermine mineral and metabolic constraints on soil carbon cycling, altering pathways and rates of carbon loss from mountainous floodplains. Global implications of our findings for predicting climate change impacts on carbon cycling within dynamic floodplains globally will be discussed.

Prof. Dr. Marco Keiluweit is interested in how biogeochemical mechanisms controlling carbon and nutrient cycles in soil and sediments respond to climate and land use change. He completed his PhD at Oregon State University, and worked as a postdoc at Stanford University and as Assistant Professor at the University of Massachusetts-Amherst. He is now Professor of Soil & Microbial Biogeochemistry in the Institute of Earth Surface Dynamics at the University of Lausanne, Switzerland. His research combines laboratory, greenhouse, and field experiments with advanced analytical tools such as synchrotron spectroscopy, chemical imaging, and molecular microbiology. His group’s work links fine-scale biogeochemical mechanisms to landscape-scale processes within natural and managed ecosystems. Prof. Keiluweit has acquired numerous grants and has received several prestigious awards, including US DOE Lawrence Scholar and NSF Early Career awards.


TUCHEM Lecture

'Emerging Two-Dimensional Materials for Environmental Applications'

Dr. Ali Shaygan Nia

Dr. Ali Shaygan Nia

Professur für Molekulare Funktionsmaterialien
TU Dresden

27.04.23, 1:30 pm in KUBUS hall 1A

 

2D materials are important building blocks for the next generation of electronic and energy devices due to their remarkable chemical and physical characteristics. They have recently shown promising performances for environmental applications such as water/air purification, anticorrosion/antiviral/antibacterial coatings, etc. To this end, large‐scale production of 2D materials with high purity and specific functionalities represents a key to advancing fundamental studies and industrial applications. Therefore, we are developing scalable wet chemistry methodologies to synthesized functionalized 2D materials.

Among different synthetic protocols, electrochemical exfoliation [1] of layered materials is a very promising approach that offers high yield, great efficiency, low cost, simple instrumentation, and excellent up‐scalability. Remarkably, playing with electrochemical parameters enables functionali-zation and tunable material properties and increases the material diversities from graphene to a broad spectrum of 2D semiconductors [2].

High solution processability of 2D materials via electrochemistry also offers hybridization/ functionalization of 2D materials and improve their processabilities into functionalized inks/paste/formulations. Wet chemistry also enables us to develop transition metal carbides/nitrides (Mxene) via safe and sustainable flouride-free approaches.


abstract fig 1 Figure 1. Schematic illustration of electrochemical exfoliation methodology and different 2D
materials which could be produced via this method

References:
[1] Adv. Mater. 2020, 32, 1907857.
[2] Adv. Mater. 2020, 32, 1907244; Small 2019, 15, 1901265; Angew. Chem. Int. Ed. 2018, 57, 4677-4681; Angew. Chem. Int. Ed. 2018, 57, 15491-15495.


AG Biophotovoltaics Open Lecture

'Engineering cyanobacteria for direct solar chemical and fuel production'

Peter Lindblad

Prof. Peter Lindblad

Microbial Chemistry
Uppsala University, Sweden

24.04.23, 10:00 am in KUBUS hall 1CD

 

Cyanobacteria, prokaryotic microorganisms with basically the same oxygenic photosynthesis as higher plants, are excellent green cell microfactories for sustainable generation of renewable chemicals and fuels from solar energy and CO2. In the presentation I will visualize the concept green cell factories by discussing two examples: (i) Generation of functional semisynthetic [FeFe]-hydrogenases linking to the native metabolism in living cells of cyanobacteria cells, and (ii) Engineering cyanobacteria to produce the bulk chemical and blend-in molecule butanol from sunlight and CO2.
[FeFe] hydrogenases are the most efficient molecular catalysts known to date with regards to H2 production. Cyanobacteria harbor native [NiFe] hydrogenase(s) but no [FeFe] hydrogenase. A heterologously expressed [FeFe] hydrogenase was synthetically activated in a hydrogenase deficient strain of the cyanobacterium Synechocystis PCC 6803 (1). H2-evolution assays showed that the non-native, semi-synthetic enzyme links to the native metabolism in living cells. The artificial activation technology was expanded to a newly discovered [FeFe]-hydrogenase which when expressed in Synechocystis showed stable expression and significant H2 production under different environmental conditions (2). This semisynthetic [FeFe]-hydrogenase remained functional in vivo in cells of Synechocystis for several days. The developed technology opens up unique possibilities to investigate not only [FeFe]-hydrogenases but also other metalloenzymes in a photosynthetic microbial cell environment, completely bypassing the many challenges of e.g. biological maturation and regulations.

Cyanobacteria lack the butanol biosynthetic pathways and corresponding relevant gene(s). Introduction of a single gene encoding KivD results in isobutanol producing strains. Further engineering and long-term experiments demonstrated a maximal cumulative isobutanol titer of 1.2 gram per liter (3, unpublished). Similarly, introducing a complete 1-butanol pathway, together with additional enzymes to increase the pool of acetyl-CoA, resulted in cells with a cumulative titer of 4.8 g per L, and a maximal rate of 600 mg 1-butanol per L and day and carbon partitioning of 60% (4-6). Present progress towards a practical system will be presented and discussed.

(1) Wegelius et al 2018. Generation of a functional, semisynthetic [FeFe]-hydrogenase in a photosynthetic microorganism. Energy & Environmental Science 11: 3163-3167

(2) Wegelius et al 2021. Semisynthetic [FeFe]-hydrogenase with stable expression and H2 production capacity in a photosynthetic microbe. Cell Reports Physical Science 2: 100376

(3) Xie & Lindblad. 2022. Effects of expressing 2-keto acid pathway enzymes on photosynthetic isobutanol production. Microbial Cell Factories 21: 17

(4) Wichmann et al 2021. Engineering Biocatalytic Solar Fuel Production: The PHOTOFUEL Consortium. Trends in Biotechnology 39: 323-327

(5) Liu et al 2021. Engineering cyanobacteria for photosynthetic butanol production. In: Photosynthesis. Biotechnological Applications with Microalgae (Ed: Rögner) 2: 33-56. Walter de Gruyter GmbH, Berlin/Boston (6) Liu et al 2022. Current advances in engineering cyanobacteria and their applications for photosynthetic butanol production. Current Opinion in Biotechnology 73: 143-150


AG Biophotovoltaics Open Lecture

'Design meets evolution: Theory and practice for fine-tuned bioengineering'

Prof. Víctor de Lorenzo

Prof. Víctor de Lorenzo

Systems Biology Department
Centro Nacional de Biotecnología, CSIC

24.04.23, 9:00 am in KUBUS hall 1CD

 

The prevailing view of biological evolution is not unlike bricolage/pastiche/tinkering—in sharp contrast with rational engineering. Yet, different paths often lead to solutions that coincide or converge whether they emerge from naturally-occurring evolution or rationally designed. Such a conjunction—often presented as a mere anecdote— in fact reveals the ability of biological systems to physically explore solution spaces and gravitate towards information-rich attractors, which can be found through different routes. This scenario evokes one of heterotic computing, a non-conventional type of data processing in which the solution to a problem is not delivered through numerical calculations but through its embodiment in a material object. Once left to undergo a physical process the object manages a large number of parameters for reaching a multi objective optimum. The course of information is thus a physical flow and the outcome is a physical currency. The consequences of this notion for bioengineering are remarkable, as it enables solutions to multi-objective optimization challenges not yet amenable to all-rational approaches. The ensuing technical question is how to bring about hyper-diversification not only of genomic sequences but also environmental and context-dependent parameters for securing the desired performance of a given synthetic device. This issue will be illustrated with a number of practical cases where naturally-occurring or artificially enhanced variability was key to find ideal outcomes to otherwise intractable design hitches of interest for industrial and environmental biotechnology. (1) Al-ramahi et al. (2021) ssDNA recombineering boosts in vivo evolution of nanobodies displayed on bacterial surfaces. Comms Biology 4: 1169. (2) Tas et al. (2020) Contextual dependencies expand the re-usability of genetic inverters. Nature Comms 12: 355.

(3) Espeso et al. (2020) An automated DIY framework for experimental evolution of Pseudomonas putida. Microb Biotechnol. 14: 2679-2685
(4) Hueso-Gil et al. (2019) Multiple-site diversification of regulatory sequences enables inter-species operability of genetic devices. ACS Synth Bio 9: 104–114.
(5) Akkaya et al. (2019) Evolving metabolism of 2,4-dinitrotoluene triggers SOS-independent diversification of host cells. Env Microbiol 21: 314-326
(6) Pérez-Pantoja et al. (2013) Endogenous stress caused by faulty oxidation reactions fosters evolution of 2,4-dinitrotoluene-degrading bacteria. PLoS Genetics 9:e1003764

IP4 Science Talks

'Achieving sustainable wastewater treatment using a green technology'

Qilin Wang

Prof. Qilin Wang

School of Civil and Environmental Engineering
University of Technology Sydney, Australia

19.04.23, 2 pm in KUBUS hall 1A

 

Wastewater treatment is essential in protecting humans and ecosystems from harmful and toxic substances in wastewater. However, the existing wastewater treatment plants (WWTPs) produce large amounts of waste sludge, accumulate antibiotic resistance genes, disperse pathogens, suffer from unsatisfactory nitrogen removal, release greenhouse gases, consume energy, and destroy the energy embedded in wastewater. This talk proposes a green free ammonia (FA) technology that could solve these issues and achieve sustainable wastewater treatment based on our ground-breaking discoveries.
The FA technology increases the volatile solids destruction of anaerobic digestion by 12-26% and improves dewaterability of anaerobically digested sludge by 9-13%, which collectively cause a volume reduction of waste sludge by 15-21%. This reduces the cost of sludge disposal. The FA technology expands the treatment capacity of anaerobic digesters by 50%, which enables the WWTPs to accommodate population growth (i.e. increasing sludge production) and save the cost for the upgrade of WWTPs. In addition, the FA technology could enhance nitrogen removal from wastewater by 30%, facilitating treated wastewater to meet regulatory discharge standards.