DOI: 10.3303/CET24109008 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Paper Received: 2 January 2024; Revised: 2 February 2024; Accepted: 8 April 2024 
Please cite this article as: Casella P., Loffredo R., Rao M.A., Balducchi R., Molino A., 2024, A Review on the Valorization of Lignocellulosic 
Biomass for Succinic Acid Production: Strengths and Weaknesses, Chemical Engineering Transactions, 109, 43-48  
DOI:10.3303/CET24109008 
  

 CHEMICAL ENGINEERING TRANSACTIONS  
 

VOL. 109, 2024 

A publication of 

 

The Italian Association 

of Chemical Engineering 

Online at www.cetjournal.it 

Guest Editors: Leonardo Tognotti, Rubens Maciel Filho, Viatcheslav Kafarov 

Copyright © 2024, AIDIC Servizi S.r.l. 

ISBN 979-12-81206-09-0; ISSN 2283-9216 

A Review on the Valorization of Lignocellulosic Biomass for 

Succinic Acid Production: Strengths and Weaknesses 

Patrizia Casellaa, Raffaele Loffredoa,b, Maria A. Raob, Roberto Balducchic, Antonio 

Molinoa,*  

aItalian National Agency for New Technologies, Energy and Sustainable Economic Development – sustainability, 

biotechnology and agroindustry division, Bioprocess and Bioproducts Laboratory (ENEA-SSPT-BIOAG-PROBIO), Piazz. le 

Enrico Fermi 1, 80055 Portici (NA), Italy 
bDepartment of Agricultural Sciences, University of Naples, Federico II, Via Università 100, 80055 Portici (NA), Italy 
c Italian National Agency for New Technologies, Energy and Sustainable Economic Development Energy and Sustainable 

Economic Development – sustainability, biotechnology and agroindustry division, Bioprocess and Bioproducts Laboratory 

(ENEA-SSPT-BIOAG-PROBIO), Strada Statale Jonica km 419 + 500, 75026 Rotondella (MT), Italy 

antonio.molino@enea.it 

Bio-based succinic acid production holds great promise for the process sustainability through the use of 

renewable resources such as lignocellulosic biomass and the use of microorganisms that can utilize CO2 in 

fermentation processes. A critical review of the latest findings on succinic acid production from lignocellulosic 

biomass was discussed, by highlighting all the key aspects of optimization processes at different levels of the 

process from pre-treatment to succinic acid production. Chemical-physical pretreatments seemed to be the 

most appropriate for the pretreatment of lignocellulosic biomass, but many strategies are still needed to solve 

the problem of variability in initial composition, the presence of post-treatment inhibitory compounds and the 

use of green solvent. In addition, the saccharization-fermentation process seemed to be the most suitable for 

the fermentative process of hydrolysates obtained from pretreatment to produce bio-based succinic acid. 

Certainly new studies and strategies are still needed to overcome the weaknesses of the whole process of 

succinic acid production from lignocellulosic biomass. 

1. Introduction 

The succinic acid is one of the most valuable bulk chemicals for many applications in food, chemical and 

pharmaceutical sectors. Nowadays, it is still synthetized chemically through the hydrogenation of petroleum-

based maleic acid or maleic anhydride. This route has been no longer sustainable for the use of fossil source 

and the environmental problems, and for the energy-intensive consumption. So, a sustainable bio-based 

production is necessary to substitute this highly impacting conventional process. 

The Bio-based production of succinic acid can be performed as intermediate in the tricarboxylic acid cycle 

(TCA) through fermentation process. Natural succinic acid producer, like the bacteria Actinobacillus 

succinogenes and Mannheimia succiniciproducens, and genetically engineered strain as Escherichia coli and 

Saccharomyces cerevisiae are able to use CO2 through fermentation for the production of succinic acid by 

reducing the carbon footprint of the process (Song & Lee, 2006). 

Although fermentation processes can be an effective alternative to chemical synthesis processes, efficient 

strategy are necessary since many difficulties needed to be overcome such as cost-effectiveness, yield and 

productivity of succinic acid (Gao et al., 2023). The valorization of lignocellulosic biomass can be a strategy to 

increase the sustainability of bio-based succinic production and to reduce waste production for new green 

products and materials but new insights are necessary to overcome the major weaknesses.  

The process of converting lignocellulosic biomass into succinic acid mainly consisted of several steps: 

pretreatments, hydrolysis, fermentation, and downstreaming process. The pretreatment and the enzymatic 

hydrolysis are fundamental preliminary steps for the availability of fermentative sugars for succinic acid 

producers strain but optimized operative conditions were necessary to convert cellulose and hemicellulose to 

43

mailto:antonio.molino@enea.it


glucose and xylose and by reducing potential unwanted compounds (Stefanidis et al., 2014). Also the 

performance of fermentation needed to be assessed and optimized to enhance succinic acid concentration, 

productivity, yield and to avoid the production of by-products. Furthermore, the downstreaming process is one 

of the most critical steps since it contributed about 50 % of the final cost of succinic acid. 

In this review the production of succinic acid from different lignocellulosic biomass were analyzed in the most 

recent works. A critical assessment was carried out on operative conditions and succinic acid production. 

Furthermore, the main challenge faced in pretreatments and processes have been discussed by highlighting 

news perspectives. 

2. Lignocellulosic biomass 

The employment of lignocellulosic biomass to produce fuels and chemicals was gaining interest over the last 

decades. These biomasses derived principally from residues/waste of various production chains in agro-

industrial sectors that were quite abundant with a production of 2 x 1011 tons per year worldwide (Kumar et al., 

2020). The most plentiful were rice, wheat, maize, and sugarcane, due to the intensive use in agriculture 

sector for food and livestock (Vivek et al., 2017).  

Since lignocellulosic biomass was potentially rich in fermentable sugars, these could be used as substrates for 

the biotransformation processes. Although lignocellulosic biomass was considered an ideal feedstock for the 

production of bio-based succinic acid, some critical issues limited their use. The principal critical issues 

concerned the variability in the composition of cellulose, hemicellulose and lignin and the consequent 

optimization of pretreatment processes.  

Before the fermentation process, a series of pre-treatments were needed to free cellulose, and hemicellulose, 

that were bounded in a complex matrix, together with lignin. Furthermore, after the pre-treatments 

carbohydrates (cellulose and hemicellulose) were broken down into their monomers, such as glucose and 

xylose, that could be used by microorganisms for the fermentation process.  

The harsh conditions of the pre-treatments, like high temperatures, and non-specificity of the process leaded 

to the formation of many by-products, such as organic acids, furans and phenolic compounds, identified as 

potential inhibitors of the growth of microorganism and consequently of the productivity of the fermentative 

processes. 

3. Pretreatments 

Lignocellulosic biomasses were a complex matrix in which the compounds contained in greatest quantities 

were cellulose (40-60% dw), hemicellulose (10-40% dw) and lignin (15-30% dw) (Lu et al., 2021). Based on 

the different feedstocks, the percentage composition of these three components may varied significantly as it 

possible to see in Table 1. 

Table 1: composition of lignocellulosic biomass from different sources (dw %). 

Lignocellulosic biomass Cellulose (%) Hemicellulose (%) Lignin (%) Reference 

Corn stalks 50 20 30 Christopher et al. (2017) 

Corn stover 38 23 20 Wan & Li (2010) 

Sugarcane bagasse 35 36 16 Sasaki et al. (2003) 

Rice straw 36 21 24 Imman et al. (2015) 

Wheat straw 40 21 23 Khan et al. (2012) 

Mischantus straw 42 25 23 Dąbkowska et al. (2019) 

 

It was possible to note that the composition of lignocellulosic biomass strongly depended on the source. The 

corn stalks had the major percentage of cellulose (50% dw) and lignin (30% dw), but the hemicellulose 

percentage (20% dw) was the lowest among the analyzed biomasses (Christopher et al., 2017). The highest 

content of hemicellulose was found in sugarcane bagasse (36% dw) (Sasaki et al., 2003). While, straw 

(wheat, rice and Mischantus) were the biomass that contained the highest percentage of lignin (> 23% dw.). 

Due to the complexity of lignocellulosic matrix, pretreatment processes were a crucial first step to make 

sugars available for the microorganisms involved in fermentation process. The pretreatment altered the 

lignocellulose structure, removed lignin, and made cellulose more accessible to enzymatic hydrolysis that 

converted the carbohydrates polymers into fermentable sugars, such as glucose and xylose. This is a 

multistep step through the synergistic action of cellulolytic and hemicellulolytic enzymes (Yang et al., 2011). 

There were developed numerous pretreatment techniques, that could be grouped into 3 different categories: 

physical, chemical, and chemical-physical.  

44



Physical methods included mechanical processes, such as grinding and ultrasound treatment that required 

high energy and high operational costs. Although another limitation to the use of physical methods was the 

low yields, Putri et al., 2023 tested peracetic and alkaline peroxide under assisted ultrasound pre-treatment on 

rice straw and bagasse by obtaining an increase of cellulose content of 107% and 68% respect to initial 

feedstock.  

Mineral acids (such as HCl, H2SO4, H2PO4, oxalic acid) and bases (calcium and NaOH and aqueous 

ammonia) and/or organic green solvent (organosolv methods) were mainly used in chemical pretreatments. 

Jampatesh et al. (2019) compared acid pretreatment on rice straw with hydrochloric, phosphoric, and sulfuric 

acid at 1M at different hydrolysis times (15, 30, 45, and 60 min) and obtained the best concentration of sugars 

(10 g/l glucose) and (18 g/L xylose) by using HCl for 60 min. Lo et al. 2020 optimized the acid pretreatment of 

sweet sorghum bagasse at 50 °C, 1 hour to obtain 29.2 cellulosic glucose after enzymatic hydrolysis. 

Chemical methods were widely used for the pretreatment of lignocellulosic biomass, the most important 

strength is the high obtained yield while the disposal of acids and bases can be a considered a weaknesses.  

The steam explosion was the most used chemical-physical pretreatment where lignocellulosic biomass is 

exposed at high temperature (160-260°C) and a pressure (5-50 atm) to a saturated steam for short times (Lee 

et al., 2003). Kuglarz et al. 2018 tested steam explosion on rapeseed straw by using H2SO4 (1% solution) in a 

batch reactor at 180 °C for 10 min to test several feedstock ratios (10, 15, 20, 25% w/v) and to test several 

enzymatic dosage (Cellulase activity). The authors obtained the best glucan recovery (98.7%) by using 20% 

w/v of feedstock and the best glucose concentration equal to 23.3 g/L and xylose concentration equal to 4.5 

g/L after the enzymatic hydrolysis (13% dosage). 

The steam explosion can be considered economically advantageous and environmental sustainable if 

compared to the techniques previously mentioned. However, despite these benefits, the harsh conditions 

(high temperature and pressure) during steam explosion generated several unwanted compounds, such 

organic acids, furans (furfural and HMF (Hydroxymethylfurfural)), and phenolic compounds that could 

negatively affect the following enzymatic hydrolysis and the succinic acid production during fermentation 

process. In fact, these compounds are known as inhibitors for the microbial growth during fermentation 

process. 

In Table 2 there were reported the major inhibitor compounds generated after the steam explosion and critical 

concentration that can compromise the succinic acid production. 

Table 2: Inhibitor compounds found in pre-treated lignocellulosic biomass and their critical concentration. 

Class Compound Microorganism employed Critical 

concentration (g/L) 

Reference 

Organic acids Acetic acid A. succinogenes 33.7 Lin et al. (2008) 

Piruvic acid  59.2  

Formic acid  10.8  

Furan derivates Hydroxymethylfurfural A. succinogenes 10 Dessie et al. (2019) 

 Furfural  6  

Phenolic 

compounds 

Vanillin Corynebacterium 

glutamicum 

0-2 Xu et al. (2015) 

 Syringaldehyde  0-2  

 

Among the inhibitors, the phenolic compounds had a strong effect at low concentration (till to 2 g/l) as 

observed by Xu et al. (2015) for vanillin and syringaldehyde that totally inhibited C. glutamicum activity. These 

inhibitors can downregulated some genes for the expression of pyruvate carboxylase and 

phosphofructokinase, that had crucial role in pathways for sugar uptake, CO2 fixation and reducing power 

supply (Xu et al., 2022). Organic acids seemed most tolerated at high concentrations till to 59.2 g/l for pyruvic 

acid and 33.7 g/l for acetic acid (Lin et al. 2008) than furan derivates as furfural and HMF. 

4. Succinic acid production starting from different lignocellulosic biomass. 

In table 3 some recent works were reported on the production of succinic acid by using lignocellulosic biomass 

as sugars feedstocks. Vallecilla-Yepez et al (2021) obtained the best SA concentration equal to 27.8 g/L at a 

yield of 0.61 g/g by using corn stover hydrolysate after liquid hot water and enzymatic hydrolysis treatment. 

The authors highlighted that the highest SA concentration was reached under the effect of sodium acetate 

buffer added in control and in corn stover hydrolysate medium. 

 

45



 

Table 3: lignocellulosic biomass used for the production of succinic acid through fermentation process. 

Pretreatments before fermentation and microorganisms employed for fermentation, sugar concentrations after 

hydrolysis (g/L), succinic acid concentration (g/L) and yield (g/gbiomass/sugars) were reported. SA = succinic acid; 

NR = not reported 

Biomass Pretreatment Microorganism Sugars 

concentration  

(g/L) 

SA 

concentration 

(g/L) 

SA 

yield 

(g/g) 

Reference 

Oil palm 

trunk 
Carboxylic acid (formic, 

oxalic, citric acids) and 

Sulphuric acid 

 

A. succinogenes 33.6 (total 

sugars) 

2.5 0.13 Bukhari et al. 

(2020) 

Oil palm 

trunk 

A. succinogenes 44.38 (total 

sugars) 

10.6 0.41 Bukhari et al. 

(2020) 

Sweet 

sorghum 

bagasse 

Phosphoric acid (5–99°C) A. succinogenes 32.75 (g x 100 

g of biomass) 

(glucose) 

17.8 0.56 Lo et al. 

(2020) 

Corn fiber Liquid hot water A. succinogenes NR 27.8 0.61 Vallecilla-

Yepez et al. 

(2021) 

Rice straw Peracetic acid (20ml/g 

biomass, 3 h 35°C), 

alkaline peroxide (20ml/g 

biomass 10 h 35°C) 

assisted by ultrasound 

 

Proteus vulgaris 20 (glucose) 3.6 0.18 Putri et al. 

(2023) 

Sugarcane 

bagasse 

Proteus vulgaris 20 (glucose) 5.1 0.25 Putri et al. 

(2023) 

Rice straw Sodium hydroxide (0.1–2.0 

M), 121°C 5 min 

Escherichia coli 100 (glucose) 32 0.32 Sawisit et al. 

(2018) 

Rice straw Acid pretreatment (4 h 

room temp.) and 

autoclaving  

E. coli 100 (glucose) 14 0.14 Jampatesh et 

al. (2019) 

Sugarcane 

bagasse 

Sodium hydroxide A. succinogenes 130 (glucose) 41 0.32 Chen et al. 

(2021) 

Mischantus 

straw 

(glycerol:water mixture and 

H2SO4) Organosolv 

A.succinogenes 44.2 (total 

sugars) 

23.5 NR Dąbkowska et 

al. (2019) 

Rapeseed 

straw 

H2SO4 (1% solution) Steam 

explosion  

S cerevisiae 23.3 g/l 

(glucose) 

9.3 0.6 Kuglarz et al. 

2018 

Wheat straw No pretreatments Fibrobacter 

succinogenes 

NR 1.5 0.05 Li et al. 

(2010) 

 

Dąbkowska et al. (2019) reached the concentration of 23.5 g/L of succinic acid after the optimization of the 

ratio of Mischantus straw hydrolysate at 75:25 v/v hydrolysate to medium mixing ratio. Sawisit et al. (2018) 

investigated the simultaneous saccharification and fermentation (SSF) at batch and fed-batch by using 

alkaline pre-treated and hydrolysed rice straw treated by using cellulose and xylanase enzymes. The authors 

achieved the highest succinic acid production under fed-batch SSF than batch SSF by using the same 

hydrolysate feedstock and by using the engineered E. coli KJ122. A novel in-situ semi- simultaneous 

saccharification and co-fermentation (SSSCF) was developed by Chen et al. 2021 that reached the SA 

concentration of 41 g/L by using undried alkali-pretreated sugarcane bagasse residuals that was enzymatically 

hydrolized and progressively in the same bottles, SA fermentation was started by adding growth medium and 

A. succinogenes inoculum (10% v/v). Lo et al. 2020 after the optimization of the acid pretreatment of sweet 

sorghum bagasse (at 50 °C, 1 hour) and starting from 29.2 g/L cellulosic glucose after enzymatic hydrolysis, 

reached the concentration of 17.8 g/L of succinic acid under strictly anaerobic condition by suppling 2.5 M 

Na2CO3 and CO2 (0.5 vvm) in a 3–L reactor. Jampatesh et al. (2019) obtained the best result on SA 

concentration 14 g/L at the yield of 0.14 g/g by using 70% of hydrolysate as feedstock for the fermentation 

process. Bukhari et al. (2020) tested different organic acid (oxalic, citric and formic acids) to pretreat oil palm 

trunk biomass. The authors observed that the pre-treatment by using oxalic acid was the best to reach the 

46



highest recovery of glucose >60% but the best results on succinic acid production was obtained when citric 

acid was used to treat the biomass.  

These recent findings on the production of succinic acid from lignocellulosic biomass have shown how 

process optimization started from characterization of the biomass composition to the optimization of the 

pretreatment process, which almost always required double pre-treatment (chemical or physical/chemical and 

enzymatic hydrolysis) except when the saccharification and co-fermentation approach was experimented. On 

the fermentation process, the aspects that seemed to have the greatest influence on the performance of 

succinic acid production were: the appropriate amount of hydrolysate as feedstock, the use of CO2 or indirect 

sources of CO2, the use of pH control buffers, and the use of engineered strains such as S. cerevisae and E. 

coli that could optimize the process compared with the use of natural SA producer strains such as A. 

succinogenes. Furthermore, among fermentation methodologies, simultaneous saccharification-fermentation 

appeared to be the most promising for succinic acid production from lignocellulosic biomass. Another critical 

aspect that requires further studies is the optimization of biomass pretreatment processes to reduce the 

production of inhibitory compounds for a better valorization of lignocellulosic biomasses through the 

optimization of the pretreatment process or by developing detoxification technologies of the pretreated. 

5. Conclusions 

The evaluations on the most recent findings highlighted how several optimizations and new strategies are still 

necessary on the production of succinic acid from lignocellulosic biomass, starting from the pre-treatment of 

the biomass due to the variable initial composition of the matrix till to the fermentation process. 

Acknowledgments 

This work was supported by The National Recovery and Resilience Plan (NRRP) - Next Generation EU – 

Mission 4, Component 2, Investiment 1.4, under the “National Research Centre for Agricultural Technologies”, 

Agritech, [grant number I63C22000350007]. 

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	A Review on the Valorization of Lignocellulosic Biomass for Succinic Acid Production: Strengths and Weaknesses