Bioresource Technology

An automated, modular system for organic waste utilization using heterotrophic alga
Galdieria sulphuraria: design considerations and sustainability
--Manuscript Draft--

Manuscript Number: BITE-D-21-08879

Article Type: Case study

Keywords: algae; Galdieria sulphuraria; waste processing; food waste; modular technology

Corresponding Author: Maximilian Julius Pahmeyer, B.Eng.

German Institute of Food Technologies
Quakenbrück, Lower Saxony GERMANY

First Author: Maximilian Julius Pahmeyer, B.Eng.

Order of Authors: Maximilian Julius Pahmeyer, B.Eng.

Shahida Anusha Siddiqui

Daniel Pleissner, Dr.

Janusz Gołaszewski, Dr.

Volker Heinz, Dr.

Sergiy Smetana

Abstract: A large amount of food is wasted and valuable compounds in many agricultural side
streams are not utilized completely. Methods to process such wastes into biomass of
defined composition automatically and in decentralized locations are lacking. Thus, this
study presents a modular design for residue utilization and continuous production of
the heterotrophic alga Galdieria sulphuraria . A life cycle and economic assessment
are carried out on the hypothetical design out to define whether the proposed system
can be ecologically and economically viable. Producing one kg of dried biomass with
the proposed system would cost 4.38 € and will be associated with 3.8 kg CO 2 eq
emitted, 69.9 MJ of non-renewable energy use, and occupation of 0.09 m 2 of organic
arable land. Sustainability is comparable to some conventional protein sources, with
further improvement foreseen through avoidance of drying.

Suggested Reviewers: Jose Viriato Coelho Vargas, Doctor in chemical engineering

Professor, Universidade Federal do Paraná: Universidade Federal do Parana
jvargas@demec.ufpr.br
Expert on LCA

Rosana de Cassia de Souza Schneider

Universidade de Santa Cruz do Sul
rosana@unisc.br
Expert on LCA

Silvia Fiore
Associate Professor, Politecnico di Torino
silvia.fiore@polito.it
Expert in assessments

Niels Thomas Eriksen, Ph.D.

Associate Professor, Aalborg University: Aalborg Universitet
nte@bio.aau.dk
Expert on Bioprocess Engineering, including Galdieria sulphuraria

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Cover Letter

Dear Sir or Madam,

We hereby confirm that we mutually agree to submit the manuscript for the article

titled “An automated, modular system for organic waste utilization using

heterotrophic alga Galdieria sulphuraria: design considerations and sustainability”. It

is the original work of the authors listed and has not been submitted to bioresource

technology previously.

Prospects for cultivation of Galdieria sulphuraria, including on residues, have been

discussed in different publications. These include laboratory and modelling studies.

The novelty of this work lays in the description and detailed analysis of a modular

system from both an ecological and economic standpoint. A comparison to

conventional sources of protein is made as well, to put the results into perspective.

Most significantly, this work illustrates the importance of a well thought-through

process to ensure a lower environmental impact than for established sources of

biomass. The analysis of different process steps reveals where the biggest impacts

lay and how a more sustainable process can be implemented. The same applies to

the economic viability of the concept. Thus, the work highlights some necessary next

steps to make the alga Galdieria sulphuraria part of a more circular economy, which

also explains the choice of the subject classification “80.020: Circular bioeconomy

and energy & environmental sustainability”.

With kind regards,

The authors
Manuscript Click here to view linked References

An automated, modular system for organic waste

utilization using heterotrophic alga Galdieria

sulphuraria: design considerations and sustainability

Maximilian Julius Pahmeyera,*, Shahida Anusha Siddiquia,b*, Daniel Pleissnerc,d,

Janusz Gołaszewskie Volker Heinza, Sergiy Smetanaa

a
German Institute of Food Technologies (DIL e.V.), Prof.-von-Klitzing-Str. 7 49610

Quakenbrück, Germany

b
Technical University of Munich Campus Straubing for Biotechnology and

Sustainability, Essigberg 3, 94315 Straubing, Germany

c
Sustainable Chemistry (Resource Efficiency), Institute of Sustainable Chemistry,

Leuphana University of Lüneburg, Universitätsallee 1, C13.203, 21335 Lüneburg,

Germany

d
Institute for Food and Environmental Research e. V., Papendorfer Weg 3, 14806 Bad

Belzig, Germany

e
Center for Bioeconomy and Renewable Energies, Department of Genetics, Plant

Breeding and Bioresource Engineering, Faculty of Agriculture and Forestry, University

of Warmia and Mazury in Olsztyn, 10-719 Olsztyn, Poland

*Corresponding authors:

E-mail address: max.pahmeyer@gmail.com postal address: Wittekindstraße 50,

33615 Bielefeld, Germany. (M. Pahmeyer)

E-mail address: s.siddiqui@dil-ev.de (S.Siddiqui)

1
Abstract

A large amount of food is wasted and valuable compounds in many agricultural

side streams are not utilized completely. Methods to process such wastes into

biomass of defined composition automatically and in decentralized locations are

lacking. Thus, this study presents a modular design for residue utilization and

continuous production of the heterotrophic alga Galdieria sulphuraria. A life

cycle and economic assessment are carried out on the hypothetical design out

to define whether the proposed system can be ecologically and economically

viable. Producing one kg of dried biomass with the proposed system would cost

4.38 € and will be associated with 3.8 kg CO2 eq emitted, 69.9 MJ of non-

renewable energy use, and occupation of 0.09 m2 of organic arable land.

Sustainability is comparable to some conventional protein sources, with further

improvement foreseen through avoidance of drying.

Keywords:

Algae, Galdieria sulphuraria, waste processing, food waste, modular technology

1 Introduction

Globally, about 1.3 billion tonnes of food, or one third of the total production, is

wasted every year (Gustavsson et al., 2011). Alongside large amounts of

agricultural side streams such as manure, digestate, straw or inedible plant

parts (ECN, 2019; Eurostat, 2018), these residues present a large source of

2
biomass. However, current practices, such as landfill, incineration, composting

or anaerobic digestion, only make use of the contained biological compounds

energetically or as plant nutrients, if at all (Gao et al., 2017). In principle, these

compounds could also be utilised as feedstock for the production of e.g. fine

chemicals, polymers or food and feed using chemical or biotechnological

processes (Ndubuisi Ezejiofor et al., 2014). However, the implementation of

such processes is hampered by unknown composition (Pleissner et al., 2017)

and decentralised production of these side streams (Favoino et al., 2020).

A much-discussed option for novel production processes is the cultivation of

microalgae, and in the context of residue utilisation, heterotrophic microalgae.

These could serve as a source of protein (Smetana et al., 2017), biodiesel (Li et

al., 2007) or fine chemicals (Kwan et al., 2015). In order to minimise the risk of

pathogen contamination, Galdieria sulphuraria, a red alga thriving at pH

between 1.5 and 2, may be a viable option (Albertano et al., 2000). This

organism is able to grow heterotrophically on a wide range of organic carbon

sources, e.g. food waste hydrolysate (Sloth et al., 2017). In addition, it has been

proposed as a food source or nutraceutical due to high content of antioxidants,

micronutrients, vitamins, fibre and protein (Graziani et al., 2013). At the same

time, little is known about the potential environmental impact and economic

costs of G. sulphuraria cultivation in heterotrophic conditions on food waste.

Recent LCA studies on cultivation of G. sulphuraria considered only carbon

footprint of autotrophic and heterotrophic cultivation systems (Thielemann et al.,

2021a) and G. sulphuraria production on food waste (Thielemann et al., 2021b).

3
Current research aims on the assessment of a hypothetical biorefinery system

which can transform organic residues into usable products of defined

properties, using heterotrophic G. sulphuraria cultivation. Modularity is a central

aspect of this system, housed within a standard 40 foot high cube shipping

container, enabling pre-assembly and location flexibility (Krasberg et al., 2014)

where residues are produced. Therefore, current manuscript presents a

sustainability assessment of the modular cultivation system from an

environmental (Life Cycle Assessment) and an economical perspective within

the geographical context of the European Union (EU).

2 Material and methods

2.1 Process modelling

The basis of the cultivation system was a 2 m3 fermenter, chosen to fit into the

container alongside other equipment. The biomass productivity and thus

required mass flow of substrate and required capacity of downstream and

upstream processing equipment were then calculated based on kinetic

equations (Takors and Weuster-Botz, 2018). Fermentation was planned to be

carried out continuously with an estimated cell density of 50 g L -1, which is

below the 83.3 g L-1 attainable in continuous fermentations in the laboratory

(Graverholt and Eriksen, 2007). For the calibration of this model, experimental

data on heterotrophic G. sulphuraria fermentation was used (Sloth et al., 2017).

At a dilution rate of 90% of the maximum specific growth rate of 0.96 d -1 (Sloth

et al., 2017), the biomass productivity could be calculated as in Eq. (A.1). is in

turn allowed calculating the glucose mass flow rate, as per Eq. (A. 2). An

4
enzymatic hydrolysis step needed for medium preparation was modelled

according to experimental data from a study assessing food waste hydrolysis

for continuous flow cultivation of heterotrophic algae (Pleissner et al., 2017).

Considering that the input hydrolysate would have to be centrifuged to separate

out solids resulted in some hydrolysate lost with the solids-rich fraction,

resulting in a food waste input needed to cover both the algae demand of

glucose and some losses with the solids fraction, as per Eq. (A. 3) the needed

volumetric flow rate of hydrolysate was calculated based on a glucose

concentration of 96 g L-1 in the hydrolysate, which also allowed calculating the

amount of water input needed for this hydrolysis including losses during

separation.

2.2 Life cycle assessment

The life cycle assessment (LCA) was carried out according to ISO 14044:2006

(ISO, 2006a) and ISO 14040:2006 (ISO, 2006b) using the SimaPro 8.0.1

software (PRé Sustainability B.V., Gouda, the Netherlands) with the background

life cycle inventory (LCI) data from ecoinvent 3.4 (ecoinvent, Zurich,

Switzerland) (Wernet et al., 2016). The life cycle impact assessment (LCIA)

method IMPACT 2002+ (Jolliet et al., 2003) was used to assess the

environmental impacts of this life cycle inventory. This approach aggregates the

impact from all input processes according to the life cycle inventory (LCI) across

fifteen impact categories. Wherever possible, inputs with the geographical

indication “Europe without Switzerland” were used. If no data for Europe

specifically was available, “GLO” for global averages or “RoW”, for global

5
averages without Switzerland was used. All impact processes taken from

ecoinvent 3 used the allocation at point of substitution (APOS) approach.

The goal of the LCA was to assess the environmental impact of the proposed

system in the context of protein production and comparing it to conventional

benchmark protein sources. The scope of the LCA was from cradle to gate, with

system boundaries shown in Error! Reference source not found.. Impacts

from all employed materials for construction and running of the module as well

as treatment of side streams such as wastewater or hydrolysis residues were

considered. The use phase of produced biomass after it leaves the module was

not considered. Under the allocational approach, 100% of the impact was

allocated to the dried algae powder (since it is considered the product and price

for economic allocation is uncertain), however, avoided impact through

substitution of waste treatment was considered. To that end, substitution of an

ecoinvent 3 process representing global average impacts of waste treatment

was considered. This process aggregates the methods incineration, composting

and anaerobic digestion according to their market share. The functional units

assessed were 1 kg of dried algae powder produced (FU 1), 1 kg of protein

produced (FU 2).

Following assumptions were included in the modelling the process:

 Maintenance and cleaning was performed twice a year for a week

(downtime of cultivation).

 System operated for 350 days of continuously per year.

 Servicing company is located 30 km distance from the module.

6
  The shredder has a capacity of 320 kg h-1

 The mass of the tanks is approximated as a hollow cylinder with 3 mm

wall thickness and a material density of 8000 kg/m3 (Thyssenkrupp

Materials (UK), 2021), with the same amount of mass added for

supports, stirrer, baffles etc. when referring to the process vessels.

 The environmental impact of production of smaller components (valves,

sensors, screws, dosing pumps etc.) is neglected

2.3 Economic assessment

The goal of the economic assessment was to estimate the specific cost of

production as well es the necessary break-even price (BEP) to make the

investment pay back within five years. The costs included capital and

operational investments as well as the treatment of waste with a profit of

0.07088 € kg-1 (including a 10% security deduction from the price used by ITES

et al. (2020)). Subtracting that profit from the specific variable cost gave the net

specific variable cost.

3 Results and Discussion

3.1 Design

The design is based on a 40 ft high cube sea container. A pipe and

instrumentation diagram is given in Fig. 2. Organic waste is put into a grinder,

which reduces its particle size. The waste is fed then into a continuously stirred

hydrolysis vessel. Water for dilution, amylolytic and proteolytic enzymes for

hydrolysis, and an appropriate amount of sulphuric acid to adjust the pH to the

7
optimum of 4.5 is added. The broth from this hydrolysis is continuously fed into

a continuous disc centrifuge, with the clear flow from this centrifuge containing

sugars and free amino acids fed into a 2 m3 bioreactor. The solids fraction is

discharged. The pH within the bioreactor is adjusted to 2 and the temperature

kept at 50° C, making contamination unlikely and enabling a continuous

process. Cultivation broth is continuously fed into a disc stack centrifuge as

well, producing a high-cell density paste. This paste is subsequently dried using

a spray dryer and can then be stored for later pick-up.

3.2 Material and energy balances

For the algal process, the proposed bioreactor concept results in algal dry

biomass production of 3.6 kg h-1. This amount requires 4.55 kg h-1 of glucose

input which, corrected for some loss of glucose with the discharged broth and

some fraction of that reused, results in 5.45 kg h-1 of glucose input needed from

the hydrolysis. Producing this amount of glucose, accounting for the losses

during separation, requires a food waste input of 16.50 kg h-1 dry mass or 55.94

kg h-1 wet mass, which means a daily input of 1342.66 kg(wm) d-1. This requires

a hydrolysis reactor with a liquid volume of at least 206.48 L. As side-streams,

22.68 L h-1 retentate slurry from the hydrolysis and 56.45 L h-1 of residual algae

culture broth from separation is produced. Furthermore, 0.1159 g h -1 of sulfuric

acid, 4.78 ml h-1 amylolytic and 2.97 L h-1 proteolytic enzyme preparation are

needed for the hydrolysis. For the bioreactor, 36.63 g h-1 H2SO4 are required to

adjust the pH and 5.32 m3 h-1 of air are necessary to keep it aerated. The stirrer

requires a power of 1 kW el to ensure sufficient aeration. The detailed life cycle

8
inventory resulting from these material flows is given in Table A. 1: life cycle

inventory for operating the module for one year, producing 19925 kg a -1 of dried

algae powder and consuming 469931.3 kg a-1 of food waste (wet mass).Table

A. 1

3.3 Life cycle assessment

The results of LCA in different impact categories are presented in Table 1. The

production of 1 kg of dried algal biomass is associated with impact on global

warming potential (GWP) in the scope of 3.48 kg CO2 eq, consumption of 69.19

MJ of non-renewable energy (NRE) and occupation of 0.09 m2 of arable land

(LU). When correcting for a protein content of 27.84% (Graziani et al., 2013) in

the dried powder to make it more comparable to other protein sources, the

impact is 12.49 kg CO2 eq, 242.72 MJ and 0.32 m2 of land respectively.

Comparing to benchmarks in the ecoinvent 3 database, this puts the

greenhouse effect below that of red meat (40.8 kg CO2 eq per kg protein) and

whey (13.47 kg CO2 eq per kg protein) but above that of tofu and chicken (7.78

and 9.1 kg CO2 eq per kg protein respectively), while energy use is higher and

land use lower than those of compared benchmarks. When considering the

consequence of avoiding waste treatment, the impact of producing one kg of

protein with the proposed system is 8.73 kg CO2 eq (GWP), 202.83 MJ NRE

used and 0.25 m2 of arable land. GWP in such case is lower than the one

outlined for chicken meat (ecoinvent, Zurich, Switzerland) (Wernet et al., 2016).

Error! Reference source not found. shows the integrated environmental

impact across impact categories, in comparison to different protein sources in

9
the ecoinvent 3 database. The aggregated impact of all sub processes taken

together (4.76 mPt) is much lower than for red meat (14.63 mPt) but slightly

higher than for whey (4.65 mPt), tofu (3.63 mPt) or chicken (4.12 mPt).

However, if the avoided impact is considered through the substitution of waste

treatment, the aggregated impact (3.34 mPt) is lower than for tofu. It should be

noted that over half the impact originates from the drying step at 2.63 mPt, due

to its high energy consumption.

Comparing the results of this study to others assessing the environmental

impact of heterotrophic algal biomass are favourable. Literature LCA results for

heterotrophic C. vulgaris cultivation on glucose using the same methodology

and database found environmental impacts of around 5 mPt kg-1 for dried algal

biomass (Smetana et al., 2017) as compared to 1.34 mPt kg-1 (not considering

substitution of waste treatment) or 0.93 mPt kg -1 (considering substitution) for

this study, suggesting that the use of food waste hydrolysates can significantly

reduce environmental impact. The same study also found a higher global

warming potential for dried algae with 4% moisture (14.7 kg CO2 eq kg-1) than

for non-meat conventional benchmark sources of protein powder, whilst this

study found a lower global warming potential per kg of dried algae of only 3.47

kg CO2 eq kg-1. In the cited study, hydrolysates as carbon source and solar

energy as electricity source were found to lead to much lower impacts (2.25 kg

CO2 eq kg-1, 0.98 mPt), illustrating the importance of carbon source and

electricity supply for sustainability (Smetana et al., 2017). A study conducting an

LCA of G. sulphuraria in different production systems also assessed

heterotrophic cultivation on glucose and fertilizer. It found a global warming

10
potential of 0.45 kg CO2 eq kg-1 for a centrifuged concentrate containing 10%

dry biomass (with higher impacts for phototrophic and mixed

photo/heterotrophic cultivation) (Thielemann et al., 2021a). When correcting to

dry mass, that amounts to about 4.5 kg CO2 eq kg-1, which is more than the

result of this study even for dried biomass (3.63 kg CO2 eq kg-1 when correcting

to dry mass). When not including impacts for drying, as if using the centrifuged

concentrate, it only amounts to 1.52 kg CO2 eq kg(dm)-1. The lower impact

found by this study is unsurprising, since the results from Thielemann et al.

(2021) indicate that 53.33% of global warming potential stems from the

upstream production of Glucose, with another 4.44% related to fertilizer

production, supporting the theory that the use of residues can substantially

lower the environmental impact of heterotrophic cultivation. Furthermore, the

results agree that energy use has a substantial impact (31.11% of total global

warming potential, second-most relevant after glucose) (Thielemann et al.,

2021a). In Table 2, a comparison of some literature results and the results of

this study is summarized.

Overall, the substantial reduction in CO2 emissions by not drying the algae of

over 50% shows that using the concentrate without drying substantially

improves the sustainability of the process. Even when not accounting for the

benefits of not having to use a different waste treatment process, The global

warming potential would be at 5.17 kg CO2 eq and thus below that of protein in

tofu (7.78 kg CO2 eq per kg protein) and energy use would be reduced to 84.52

MJ primary per kg protein and thus be below that of most benchmarks, except

chicken meat (75. 86 MJ primary)

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3.4 Economic assessment

The cost of module construction and operation are summarized in Table 3. The

investment cost of 964044.28 € results in annual fixed costs of 48202.21 €,

whilst the variable cost resulting from use of acid, enzymes, electricity, water

and wastewater as well as labor for maintenance comes to 83947.96 € a-1. To

cover these costs, a price of 4.38 € kg-1 of dried biomass would be required. If

the income from waste treatment of 70.88 € t-1 is considered, 3.27 € kg-1 would

be needed to cover the cost. However, for the module to be paid back after 5

years, 8.10 € kg-1 plus the projected revenue from waste treatment would be

needed.

It should be kept in mind that these prices are for dried biomass and when, for

instance, using the biomass in extrusion process, up to 40% water may be

added (Neder-Suárez et al., 2016), reducing the price related to the input of

biomass from the modules to 4.94 € kg-1 of end product. When thinking about

other utilisation options, these prices, albeit high, may not be an issue. One

example is phycocyanin production; food grade phycocyanin produced from

spirulina exceeds prices of 200 € kg-1 even for large orders (Gewürzland GmbH,

2021) with analytical grade phycocyanin selling at prices above 100 € mg-1

(AAT Bioquest Inc, 2021; Merck KGaA, 2021). Assuming a phycocyanin

concentration of 2 mg g(dw)-1 (Sloth et al., 2006), the concentration in the end

product would be 1.92 g per kg dried algal biomass. Per kg of dried algae

powder, this would result in a value of 3.84 € kg-1 for food grade and 192000 €

kg-1 for analytical grade phycocyanin. Under optimal conditions, phycocyanin

12
content can reach up to 11.52 mg g(dw)-1 in heterotrophically grown G.

sulphuraria (Sloth et al., 2006). In this case, one kg dried algal biomass would

contain 26.88 g kg-1 of phycocyanin. This would result in a value of 5.38 € kg-1

for food grade and 2688000 € kg-1 for analytical grade phycocyanin. It should be

kept in mind that especially the production of analytical grade phycocyanin

involves a lot of costly processing steps, even more so for heterotrophic algae

with a lower total phycocyanin content than phototrophic ones (Sørensen et al.,

2013). One possibility may be to increase the phycocyanin content using

downstream phototrophic cultivation (Wan et al., 2016).

Utilisation of the hydrolysation residue, which is a lipid-rich biomass, may also

be useful. Prospectively, further amylolytic and proteolytic hydrolysis may gain a

lipid-rich biomass suitable for biodiesel production. Further additional utilisation

options include co-production of e.g. plasticizer (Kwan et al., 2015; Pleissner

and Smetana, 2020), which has been demonstrated to enhance profitability.

Using all the proteins and carbohydrates in G. sulphuraria biomass may thus

deliver a perspective for an economically viable concept.

On the other hand, phycocyanin extraction (just like some other utilisation

options) does not require drying. It may be attractive to leave out the drying

process altogether. The cost savings from the dryer investment as well as the

energy savings would reduce the necessary BEP to 4.75 € kg-1 (for the same

production volume as for dried biomass). This would be lower than the value of

the phycocyanin in the biomass even for low price estimates if the process were

optimised for its production, which may require the addition of lights to stimulate

phycobiliprotein production. Considering that the paste produced from

13
centrifugation, which has a dry biomass concentration of 22% and thus more

water, would now be in the end product (of which a total of 130581 kg is

produced in a year). The BEP would then be 1.09 €/kg wet mass.

4 Conclusions

The modular production of protein from food waste using Galdieria sulphuraria

can be more sustainable than established methods, depending on the

processing concept. Whilst dried algae powder is comparable in terms of

sustainability to some plant- and animal-based benchmarks, a real improvement

can best be achieved when aiming for a high-moisture product with avoidance

of energy-consuming drying. However, profitability is challenging due to

significant investment and operational costs. Outstanding research includes

development and assessment of a complete process from hydrolyzation and

fermentation up to extraction of high-value products.

Funding

This research is funded by the German Federal Ministry of Education and

Research (BMBF), in the frame of FACCE-SURPLUS/FACCE-JPI project

UpWaste, grant number 031B0934A and 031B0934B and by the National

(Polish) Centre for Research and Development (NCBiR) (Project FACCE

SURPLUS/III/UpWaste/02/2020).

14
Figure captions

Fig. 1. Scheme of system boundaries of heterotrophic cultivation of G.

sulphuraria on food waste; Process steps are represented with rectangles,
inputs with white circles, outputs with black circles and the system
boundary is outlined with a dashed line
Fig. 2. Pipe and instrumentation diagram of G. sulphuraria cultivation.
Prefix M: major equipment pieces, prefix E: other devices, prefix I:
measuring points, prefix P: pipes.
Fig. 3. Integrated impacts of production of 1 kg of protein using the
proposed module in comparison to conventional products corrected for
protein content. Sub-processes: Impacts according to material and energy
consumption of process steps. Substitution: avoided impact through
avoidance of waste treatment (global average). 1 kPt equal to average
annual impact of a European citizen. GS – G. sulphuraria cultivation and
processing Methodology: IMPACT 2002+.

15
Tables

Table 1. Environmental impact of 1 kg dried G. sulphuraria biomass

production Methodology: IMPACT 2002+.
Impact category Unit FU1: 1 kg of dried FU2: 1 kg of
biomass protein
No sub Sub No sub Sub

Carcinogens g C2H3Cl eq 24.97 8.59 89.69 30.87

±4.76 ±4.68 ±17.10 ±16.80
Non-carcinogens kg C2H3Cl eq 0.09 -0.01 0.31 -0.03
±0.04 ±0.04 ±0.15 ±0.16
Respiratory inorganics g PM2.5 eq 4.16 2.60 14.95 9.35
0.19 0.21 0.67 0.76
Ionizing radiation Bq C-14 eq 126.81 172.14 455.49 618.34
±88.91 ±248.78 ±319.37 ±893.59
Ozone layer depletion mg CFC-11 eq 0.36 0.27 1.30 0.97
±0.05 ±0.03 ±0.17 ±0.12
Respiratory organics g C2H4 eq 0.53 0.02 1.91 0.06
±0.05 ±0.04 ±0.16 ±0.15
Aquatic ecotoxicity kg TEG water 298.69 199.59 1072.88 716.93
±42.84 ±36.32 ±153.88 ±130.45
Terrestrial ecotoxicity kg TEG soil 87.91 68.03 315.78 244.38
±10.17 ±9.65 ±36.52 ±34.65
Terrestrial acid/nutri kg SO2 eq 0.14 0.07 0.51 0.24
±0.01 ±0.01 ±0.04 ±0.03
Land occupation m2org.arable 0.09 0.07 0.32 0.25
±0.01 ±0.01 ±0.05 ±0.04
Aquatic acidification g SO2 eq 36.36 19.61 130.59 70.43
±2.21 ±1.56 ±7.94 ±5.61
Aquatic eutrophication g PO4 P-lim 1.21 0.96 4.35 3.44
±0.69 ±0.87 ±2.49 ±3.13
Global warming kg CO2 eq 3.48 2.43 12.49 8.73
±0.10 ±0.09 ±0.35 ±0.32
Non-renewable energy MJ primary 69.19 56.47 248.54 202.83
±8.51 ±6.97 ±30.58 ±25.04
Mineral extraction MJ surplus 0.13 0.12 0.47 0.42
±0.02 ±0.02 ±0.06 ±0.06
Note: No sub - all impact allocated to dried algae powder as a main product,
no avoidance of waste treatment is considered; Sub - substitution of waste
treatment is included ; FU – functional unit; ± 4.76 – standard deviation.

16
Table 2. Comparison of integrated environmental impact and global
warming potential to benchmark scores

Organism Functional Integrated Global Reference

unit impact d, warming
mPt kg-1 potential,
kg CO2 eq
kg-1
C. vulgaris 1 kg dried 5a 14.7 a (Smetana et
biomass 0.98 b, c 2.25 b, c al., 2017)
G. sulphuraria 1.34 b 3.47 b this study
1 kg dry - 4.5 a (Thielemann
biomass in et al., 2021a)
centrifuged - 1.52 b this study
algae paste
Note: a using Glucose, fertiliser and network electricity, b using residues as

substrate, c using photovoltaic electricity, d methodology: IMPACT 2002+.

17
Table 3. Results of the economic assessment of the proposed module

Unit G. sulphuraria
Investment cost € 964,044.28
Total fixed cost (discount) € a-1 48,202.21
Total variable cost € a-1 82,947.96
Specific fixed cost € kg-1 1.61
Specific variable cost € kg-1 2.77
Specific cost € kg-1 4.38
Net specific variable cost € kg-1 1.66
Net total specific cost € kg-1 3.27
Break-even price € kg-1 8.10

18
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Appendices

Eq. (A. 1): 𝒎̇𝒙 = 𝑫 ∙ 𝑽𝑳 ∙ 𝒄𝒙

𝑚̇𝑥 dry biomass flow rate over time 𝑘𝑔 ∙ 𝑠 −1

𝑉𝐿 total liquid reactor volume 𝑚3

𝑐𝑥 dry biomass concentration in broth 𝑘𝑔 ∙ 𝑚−3

𝟏
Eq. (A. 2): 𝒎̇𝑮𝒍𝒄,𝒙 = 𝒎̇𝑫𝑩𝑴 ∙ 𝒀𝒙/𝑮𝒍𝒄

𝑚̇𝐺𝑙𝑐,𝑥 Glucose mass flow rate needed for algae growth 𝑘𝑔 ∙ 𝑠 −1

𝑌𝑥/𝐺𝑙𝑐 Biomass yield on the mass of glucose consumed 𝑘𝑔 ∙ 𝑘𝑔−1

𝒎̇𝑮𝒍𝒄,𝒉𝒚𝒅 + (𝒄𝑮𝒍𝒄,𝒉𝒚𝒅 ∙ 𝑽̇𝒓𝒆𝒕 ∙ 𝜽𝒓𝒆𝒕 )

Eq. (A. 3): 𝒎̇𝒔,𝒅𝒎 = 𝒀𝑮𝒍𝒄/𝑭𝑾

𝑚̇𝐺𝑙𝑐,ℎ𝑦𝑑 Glucose mass flow rate from hydrolysate 𝑘𝑔 ∙ 𝑠 −1

𝑚̇𝑠,𝑑𝑚 dry mass consumption rate of substrate 𝑘𝑔 ∙ 𝑠 −1

𝑐𝐺𝑙𝑐,ℎ𝑦𝑑 Glucose concentration in hydrolysate 𝑘𝑔 ∙ 𝑚−3

𝑉̇𝑟𝑒𝑡 Volumetric flow rate of retentate 𝑚3 ∙ 𝑠 −1

𝜃𝑟𝑒𝑡 liquid fraction of retentate 𝑚3 ∙ 𝑚−3

25
Table A. 1: life cycle inventory for operating the module for one year,
producing 19925 kg a-1 of dried algae powder and consuming 469931.3 kg
a-1 of food waste (wet mass).

Materials/fuels amount unit comment

a. cleaning
Tap water {Europe without Switzerland}| market
for | APOS, U 13238,89 kg
Sodium hydroxide, without water, in 50% solution in alkaline
state {GLO}| market for | APOS, U 242,92 kg detergent
Potassium hydroxide {GLO}| market for | APOS, in alkaline
U 48,584 kg detergent
Phosphoric acid, industrial grade, without water,
in 85% solution state {GLO}| market for | APOS, in acidic detergent,
U 288,42 kg 50% content
in acidic detergent,
Citric acid {GLO}| market for | APOS, U 24,51 kg 5% content
Polyethylene, high density, granulate {GLO}|
market for | APOS, U 43,15 kg
Wastewater, average {Europe without
Switzerland}| market for wastewater, average |
APOS, U 13,58 m3
b. container and land use
Occupation, industrial area 59,44 m2a in water
Steel, low-alloyed, hot rolled {GLO}| market for |
APOS, U 191 kg
Polyurethane, rigid foam {GLO}| market for |
APOS, U 74 kg
c. pumping, process control and lights
Chromium steel pipe {GLO}| market for | APOS,
U 7,2 kg pumps
Chromium steel pipe {GLO}| market for | APOS,
U 6,25 kg pipes
Electronics, for control units {GLO}| market for |
APOS, U 0,2 kg computer
Electronics, for control units {GLO}| market for |
APOS, U 0,05 kg lights
Electricity, low voltage {Europe without
Switzerland}| market group for | APOS, U 429,65 kWh pumps
Electricity, low voltage {Europe without
Switzerland}| market group for | APOS, U 1,58 kWh lamps
Electricity, low voltage {Europe without
Switzerland}| market group for | APOS, U 1260 kWh computer
d. storage
Chromium steel pipe {GLO}| market for | APOS,
U 7,57 kg paste tank
Polyethylene, low density, granulate {GLO}|
market for | APOS, U 5,45 kg paloxes
1. grinding, dilution, feeding
Tap water {Europe without Switzerland}| market density of 1000
for | APOS, U 135473,56 kg kg/m3
Chromium steel pipe {GLO}| market for | APOS,
U 16 kg shredder

26
Chromium steel pipe {GLO}| market for | APOS,
U 6,5 kg screw feeder
Electricity, low voltage {Europe without
Switzerland}| market group for | APOS, U 4405,61 kWh shredder
Electricity, low voltage {Europe without
Switzerland}| market group for | APOS, U 160,33 kWh screw feeder
2. hydrolysis
Chromium steel pipe {GLO}| market for | APOS,
U 3,92 kg vessel
Steel, low-alloyed, hot rolled {GLO}| market for |
APOS, U 6,4 kg heat exchanger
Sulfuric acid {GLO}| market for | APOS, U 0,97365 kg
sulfuric acid
Polyethylene, low density, granulate {GLO}| packaging (0.3% of
market for | APOS, U 0,02328 kg total)
Polyethylene, low density, granulate {GLO}| enyzmes
market for | APOS, U 2,87 kg packaging
Enzymes {GLO}| market for enzymes | APOS, U 40,19 kg Glucoamylase AN
Enzymes {GLO}| market for enzymes | APOS, U 24,94 kg Protease S-02
Electricity, low voltage {Europe without
Switzerland}| market group for | APOS, U 1253,19 kWh
Electricity, low voltage {Europe without
Switzerland}| market group for | APOS, U 6720 kWh
3. hydrolysate separation
Chromium steel pipe {GLO}| market for | APOS,
U 3,75 kg
Electricity, low voltage {Europe without
Switzerland}| market group for | APOS, U 1259,21 kWh
Biowaste {RoW}| treatment of biowaste, industrial
composting | APOS, U 190551,3 kg
Biowaste {RoW}| market for | APOS, U 190551,3 kg
4. fermentation
Air 53797 kg in air
Chromium steel pipe {GLO}| market for | APOS,
U 22 kg bioreactor
Steel, low-alloyed {GLO}| market for | APOS, U 19 kg
Steel, low-alloyed {GLO}| market for | APOS, U 0,3 kg
Polyethylene, low density, granulate {GLO}|
market for | APOS, U 7,79 kg
Sulfuric acid {GLO}| market for | APOS, U 307,723 kg
Electricity, low voltage {Europe without
Switzerland}| market group for | APOS, U 8400 kWh stirrer
Electricity, low voltage {Europe without
Switzerland}| market group for | APOS, U 12600 kWh heat exchanger
Electricity, low voltage {Europe without cooling circulation
Switzerland}| market group for | APOS, U 6366 kWh pump
Electricity, low voltage {Europe without
Switzerland}| market group for | APOS, U 546 kWh compressor
Carbon dioxide, biogenic 7046,83 kg

27
 5. broth separation
Chromium steel pipe {GLO}| market for | APOS,
U 3,75 kg
51.38 percent of
energy
consumption for
Electricity, low voltage {Europe without both centrifuges in
Switzerland}| market group for | APOS, U 1330,52 kWh total
Wastewater, average {Europe without
Switzerland}| market for wastewater, average |
APOS, U 251,236 m3
6. spray drying
Chromium steel pipe {GLO}| market for | APOS,
U 58,07 kg
Electricity, low voltage {Europe without
Switzerland}| market group for | APOS, U 134223,16 kWh
steam from
Water 7612,92 kg evaporation
f. transport
Transport, passenger car, large size, diesel,
EURO 5 {GLO}| market for | APOS, U 1560 km

28
Figure 1 Click here to access/download;Figure;Fig_1.jpg
Figure 2 Click here to access/download;Figure;Fig_2.jpg
Figure 3 Click here to access/download;Figure;Fig_3.jpg
Declaration of Interest Statement

Declaration of competing interest

The authors declare no competing interests.