Lignocellulosic Hydrolysates Research Articles

Acetate production from corn stover hydrolysate using recombinant Escherichia coli BL21 (DE3) with an EP-bifido pathway

BackgroundAcetate is an important chemical feedstock widely applied in the food, chemical and textile industries. It is now mainly produced from petrochemical materials through chemical processes. Conversion of lignocellulose biomass to acetate by biotechnological pathways is both environmentally beneficial and cost-effective. However, acetate production from carbohydrate in lignocellulose hydrolysate via glycolytic pathways involving pyruvate decarboxylation often suffers from the carbon loss and results in low acetate yield.ResultsEscherichia coli BL21 (DE3) was confirmed to have high tolerance to acetate in this work. Thus, it was selected from seven laboratory E. coli strains for acetate production from lignocellulose hydrolysate. The byproduct-producing genes frdA, ldhA, and adhE in E. coli BL21 (DE3) were firstly knocked out to decrease the generation of succinate, lactate, and ethanol. Then, the genes pfkA and edd were also deleted and bifunctional phosphoketolase and fructose-1,6-bisphosphatase were overexpressed to construct an EP-bifido pathway in E. coli BL21 (DE3) to increase the generation of acetate from glucose. The obtained strain E. coli 5K/pFF can produce 22.89 g/L acetate from 37.5 g/L glucose with a yield of 0.61 g/g glucose. Finally, the ptsG gene in E. coli 5K/pFF was also deleted to make the engineered strain E. coli 6K/pFF to simultaneously utilize glucose and xylose in lignocellulosic hydrolysates. E. coli 6K/pFF can produce 20.09 g/L acetate from corn stover hydrolysate with a yield of 0.52 g/g sugar.ConclusionThe results presented here provide a promising alternative for acetate production with low cost substrate. Besides acetate production, other biotechnological processes might also be developed for other acetyl-CoA derivatives production with lignocellulose hydrolysate through further metabolic engineering of E. coli 6K/pFF.

Engineering a xylose fermenting yeast for lignocellulosic ethanol production.

Lignocellulosic ethanol is produced by yeast fermentation of lignocellulosic hydrolysates generated by chemical pretreatment and enzymatic hydrolysis of plant cell walls. The conversion of xylose into ethanol in hydrolysates containing microbial inhibitors is a major bottleneck in biofuel production. We identified sodium salts as the primary yeast inhibitors, and evolved a Saccharomyces cerevisiae strain overexpressing xylose catabolism genes in xylose or glucose-mixed medium containing sodium salts. The fully evolved yeast strain can efficiently convert xylose in the hydrolysates to ethanol on an industrial scale. We elucidated that the amplification of xylA, XKS1 and pentose phosphate pathway-related genes TAL1, RPE1, TKL1, RKI1, along with mutations in NFS1, TRK1, SSK1, PUF2 and IRA1, are responsible and sufficient for the effective xylose utilization in corn stover hydrolysates containing high sodium salts. Our evolved or reverse-engineered yeast strains enable industrial-scale production of lignocellulosic ethanol and the genetic foundation we uncovered can also facilitate transfer of the phenotype to yeast cell factories producing chemicals beyond ethanol.

Synthetic biology approaches to improve tolerance of inhibitors in lignocellulosic hydrolysates

Synthetic biology approaches to improve tolerance of inhibitors in lignocellulosic hydrolysates

Engineering and evolution of Yarrowia lipolytica for producing lipids from lignocellulosic hydrolysates

Yarrowia lipolytica, an oleaginous yeast, shows promise for industrial fermentation due to its robust acetyl-CoA flux and well-developed genetic engineering tools. However, its lack of an active xylose metabolism restricts the conversion of cellulosic sugars to valuable products. To address this, metabolic engineering, and adaptive laboratory evolution (ALE) were applied to the Y. lipolytica PO1f strain, resulting in an efficient xylose-assimilating strain (XEV). Whole-genome sequencing (WGS) of the XEV followed by reverse engineering revealed that the amplification of the heterologous oxidoreductase pathway and a mutation in the GTPase-activating protein gene (YALI0B12100g) might be the primary reasons for improved xylose assimilation in the XEV strain. When a sorghum hydrolysate was used, the XEV strain showed superior xylose consumption and lipid production compared to its parental strain (X123). This study advances our understanding of xylose metabolism in Y. lipolytica and proposes effective metabolic engineering strategies for optimizing lignocellulosic hydrolysates.

Ashbya gossypii as a versatile platform to produce sabinene from agro-industrial wastes

BackgroundAshbya gossypii is a filamentous fungus widely utilized for industrial riboflavin production and has a great potential as a microbial chassis for synthesizing other valuable metabolites such as folates, biolipids, and limonene. Engineered strains of A. gossypii can effectively use various waste streams, including xylose-rich feedstocks. Notably, A. gossypii has been identified as a proficient biocatalyst for producing limonene from xylose-rich sources. This study aims to investigate the capability of engineered A. gossypii strains to produce various plant monoterpenes using agro-industrial waste as carbon sources.ResultsWe overexpressed heterologous terpene synthases to produce acyclic, monocyclic, and bicyclic monoterpenes in two genetic backgrounds of A. gossypii. These backgrounds included an NPP synthase orthogonal pathway and a mutant erg20F95W allele with reduced FPP synthase activity. Our findings demonstrate that A. gossypii can synthesize linalool, limonene, pinene, and sabinene, with terpene synthases showing differential substrate selectivity for NPP or GPP precursors. Additionally, co-overexpression of endogenous HMG1 and ERG12 with heterologous NPP synthase and terpene synthases significantly increased sabinene yields from xylose-containing media. Using mixed formulations of corn-cob lignocellulosic hydrolysates and either sugarcane or beet molasses, we achieved limonene and sabinene productions of 383 mg/L and 684.5 mg/L, respectively, the latter representing a significant improvement compared to other organisms in flask culture mode.ConclusionsEngineered A. gossypii strains serve as a suitable platform for assessing plant terpene synthase functionality and substrate selectivity in vivo, which are crucial to understand monoterpene bioproduction. The NPP synthase pathway markedly enhances limonene and sabinene production in A. gossypii, achieving levels comparable to those of other industrial microbial producers. Furthermore, these engineered strains offer a novel approach for producing monoterpenes through the valorization of agro-industrial wastes.

Evaluation of a phenolic-tolerant Wickerhamomyces anomalus strain for efficient ethanol production based on omics analysis

Lignocellulosic biomass is a highly economically viable substrate for ethanol fermentation, but the presence of phenolic compounds (PCs) in lignocellulosic hydrolysates severely hinders the growth of ethanologenic strains. This study aims to investigate the ethanol fermentation capabilities and PC tolerance of the newly isolated Wickerhamomyces anomalus CAP5. The strain demonstrated remarkable ethanol production, achieving 87.1 g/L with a yield of 0.47 g/g, exceeding 90 % of the theoretical maximum. To elucidate the mechanisms responsible for this high ethanol yield, comprehensive genome and transcriptome analyses were performed. These analyses identified key metabolic pathways, including acetate reabsorption and an incomplete glycerol synthesis pathway, as crucial factors contributing to the strain's superior performance. Moreover, W. anomalus CAP5 demonstrated notable resistance to PCs, particularly phenolic acids. In fermenting with coffee husk hydrolysate (CHH), which is rich in PCs, W. anomalus CAP5 achieved noteworthy ethanol production of 60.5 g/L, marking a record for ethanol production from CHH. Hence, W. anomalus CAP5 emerges as a promising strain for effective ethanol production from lignocellulosic hydrolysates enriched with PCs.

Xylitol production by Barnettozyma populi Y-12728 with different immobilization strategies

One of the new xylitol producer microorganisms is Barnettozyma populi Y-12728 and it has great potential for the industry with its pure xylitol production capability. Different immobilization strategies, the usability of baffled or normal flasks with different agitation speeds, and various lignocellulosic hydrolysates were studied in this research. The highest xylitol production and yield values were 11.99 g xylitol/L and 40.28 % for the C1 trial at 70 ml medium with a suspended cell. For the immobilization strategy, 1 % polyethyleneimine concentration, 1.5 mm surface lattice thicknesses, and 8 3D cubes were determined to be the optimum conditions with 17.84 g/L xylitol production and 0.473 g xylitol/g xylose yield values in a 70 ml volume medium at 200 rpm, 30 °C, and 6.0 initial pH for 3 days. Rice husk, wheat bran, and oat husk hydrolysates were also used as a substrate for xylitol fermentation. The highest xylitol production was 2.26 g/L for lignocellulosic hydrolysates. In this research, FDM (Fused Deposition Modelling) based 3D printed cubes are used for the immobilization agent of Barnettozyma populi NRRL Y-12728 for the first time. The results revealed that FDM-based 3D-printed cubes could be used to immobilize cells and improve productivity for xylitol production.

Investigation of hydrolysis of sugarcane bagasse into glucose over solid acid HSO3-ZSM-5 zeolite for fermentation to bioethanol

ABSTRACT In the pursuit of sustainable energy sources, biomass has emerged as a promising alternative to fossil fuels due to its renewability and carbon neutrality. This current work presents the application of solid acid zeolite catalyst for biomass hydrolysis into reducing sugar for fermentation. Sulfonated ZSM-5 zeolite was prepared by grafting sulfonic acid group on the surface and/or into the pores of porous ZSM-5 zeolite and characterized by several analyses. The as-synthesized HSO3-ZSM-5 zeolite was then investigated as a solid acid catalyst for the hydrolysis of alkaline pretreated sugarcane bagasse (SB). Containing sulfonic acid groups – a very strong acid – on the surface, HSO3-ZSM-5 zeolite presented high catalytic activity and high efficiency in the hydrolysis of SB biomass. When the hydrolysis was carried out under optimal conditions, approximately 106.8 g/L of reducing sugar including of glucose, xylose, arabinose, and cellobiose was recovered. 86.65% fermentable sugars could be achieved after catalytic hydrolysis, and 48.7% of SB in total was converted to reducing sugar and others. Moreover, this solid acid catalyst can be reused impressively several times with no significant loss in catalytic activity. The sugar hydrolyzate could be used for fermentative production of bio-ethanol. Results indicated the possibility of using solid acid zeolite in the hydrolysis of lignocellulose to generate sugars that can be further applied in biofuel or chemical manufacturing.

Biohydrogen production from lignocellulosic hydrolysate: Unveiling the synergistic impact of substrate concentration and furfural inhibition.

Lignocellulosic biomass offers substantial potential as an ideal feedstock for dark fermentative hydrogen production due to its sustainability and cost-effectiveness. The current study examined the influence of furfural on fermentative hydrogen production using lignocellulosic hydrolysate in the presence of furfural. Synthetic lignocellulosic hydrolysate, consisting primarily of 76% xylose, 10% glucose, 9% arabinose, and a mixture of other sugars such as galactose and mannose (85% pentose sugars and 15% hexose sugars), was employed as the substrate. Various substrate concentrations ranging from 2 to 32g/L were tested, along with furfural concentrations of 0, 1, and 2g/L. The investigation aimed to assess the effects of initial substrate concentration, initial furfural concentration, furfural-to-biomass ratio (F/B), and furfural-to-substrate ratio (F/S) on biohydrogen production yields. The maximum specific substrate utilization rates at different substrate concentrations were effectively characterized using Haldane's substrate inhibition model. Among the tested concentrations, the 16g/L emerged as the optimal substrate concentration. The initial furfural concentration was identified as the most significant parameter impacting biohydrogen production, with complete inhibition observed at a furfural concentration of 2g/L. Higher F/S ratios at substrate concentrations ranging from 2 to 16g/L resulted in reduced maximum specific hydrogen production rates (MSHPR) and hydrogen yields. Substrate inhibition was observed at 24g/L and 32g/L. Lactate was the predominant metabolite in all batches containing 2-g/L furfural, as well as in batches with 1-g/L furfural and substrate concentrations of 24 and 32g/L. Furfural at a concentration of 1g/L was not inhibitory in any of the batches. Overall, the mixed cultures in this study could efficiently produce hydrogen from lignocellulosic hydrolysates and degrade furfural, providing new insights into fermentative hydrogen-producing bacteria with furfural tolerance.

Synthesis of β-ionone from xylose and lignocellulosic hydrolysate in genetically engineered oleaginous yeast Yarrowia lipolytica.

β-ionone, an apocarotenoid derived from a C40 terpenoid has an intense, woody smell and a low odor threshold that has been widely used in as an ingredient in food and cosmetics. Yarrowia lipolytica is a promising host for β-ionone production because of its oleaginous nature, its ability to produce high levels of acetyl-CoA (an important precursor for terpenoids), and the availability of synthetic biology tools to engineer the organism. In this study, β-carotene-producing Y. lipolytica strain XK17 was employed for β-ionone biosynthesis. First, we explored the effect of different sources of carotenoid cleavage dioxygenase (CCD) genes on β-ionone production. A high-yielding strain rUinO-D14 with 122mg/L of β-ionone was obtained by screening promoters combined with rDNA mediated multi-round iterative transformations to optimize the expression of the CCD gene of Osmanthus fragrans. Second, to further develop a high-level production strain for β-ionone, we optimized key genes in the mevalonate pathway by multi-round iterative transformations mediated by non-homologous end joining, combined with a protein tagging strategy. Finally, the introduction of a heterologous oxidoreductase pathway enabled the engineered Y. lipolytica strain to use xylose as a sole carbon source and produce β-ionone. In addition, the potential for use of lignocellulosic hydrolysate as the carbon source for β-ionone production showed that the NHA-A31 strain had a high β-ionone productivity level. This study demonstrates that engineered Y. lipolytica can be used for the efficient, green and sustainable production of β-ionone.

Sustainable Production of Shinorine from Agricultural Wastes Using Engineered Saccharomyces cerevisiae Expressing Novel d-Alanine-d-alanine Ligase from Pseudonocardia pini.

Shinorine, a compound known for its protective properties against UV radiation, is widely used in cosmetics and pharmaceuticals. Despite the construction of various recombinant Saccharomyces cerevisiae strains for shinorine production, achieving industrial-scale yields remains a challenge. In this study, genes encoding enzymes (DDGS, O-MT, and ATP-grasp enzyme) from Actinosynnema mirum were introduced into S. cerevisiae DXdT to enable the heterologous conversion of sedoheptulose 7-phosphate to mycosporine-glycine─the direct biosynthetic precursor of shinorine. Subsequently, a novel d-alanine-d-alanine ligase from Pseudonocardia pini was introduced to produce shinorine. The engineered strain (DXdT-MG-mi89-PP.ddl) produced 267.9 mg/L shinorine with a 48.6 mg/g dry cell weight (DCW) content in a medium supplemented with lignocellulosic hydrolysate derived from rice straw. Notably, the recombinant strain produced 1.7 g/L shinorine with a 79.1 mg/g DCW content from a corn steep liquor medium with a mixture of glucose and xylose. These results support the idea that sustainable shinorine production from agricultural wastes holds significant promise for industrial applications.

Microbial production of levulinic acid from glucose by engineered Pseudomonas putida KT2440

Levulinic acid(LA) is produced through acid-catalyzed hydrolysis and dehydration of lignocellulosic biomass. It is a key platform chemical used as an intermediate in various industries including biofuels, cosmetics, pharmaceuticals, and polymers. Traditional LA production uses chemical conversion, which requires high temperatures and pressures, strong acids, and produces undesirable side reactions, repolymerization products, and waste problems Therefore, we designed an integrated process to produce LA from glucose through metabolic engineering of Pseudomonas putida KT2440. As a metabolic engineering strategy, codon optimized phospho-2-dehydro-3-deoxyheptonate aldolase (AroG), 3-dehydroshikimate dehydratase (AsbF), and acetoacetate decarboxylase (Adc) were introduced to express genes of the shikimate and β-ketoadipic acid pathways, and the 3-oxoadipate CoA-transferase (pcaIJ) gene was deleted to prevent loss of biosynthetic intermediates. To increase the accumulation of the produced LA, the lva operon encoding levulinyl-CoA synthetase (LvaE) was deleted resulting in the high LA-producing strain P. putida HP203. Culture conditions such as medium, temperature, glucose concentration, and nitrogen source were optimized, and under optimal conditions, P. putida HP203 strain biosynthesized 36.3 mM (4.2 g/L) LA from glucose in a fed-batch fermentation system. When lignocellulosic biomass hydrolysate was used as the substrate, this strain produced 7.31 mM of LA. This is the first report of microbial production of LA from glucose by P. putida. This study suggests the possibility of manipulating biosynthetic pathway to produce biological products from glucose for various applications.

Corn-derived Expansin synergistically promotes enzymatic hydrolysis of corn cob

The enzymatic hydrolysis of lignocellulose is hindered by challenges such as high enzyme usage and associated costs. It is essential to explore effective approaches to improve the efficiency of enzymatic hydrolysis while reducing costs. Expansins are non-enzymatic proteins that can interact with lignocellulose and facilitate the loosening of plant cell walls. Given their natural affinity to plant cell walls, we hypothesized that a corn Expansin could enhance the enzymatic hydrolysis of corn lignocellulose. In this study, we expressed a corn (Zea mays) Expansin, EXPA17, in yeast cells and explored the synergistic effect between EXPA17 and commercial cellulase, and found that EXPA17 exhibited a pronounced synergistic effect on the enzymatic hydrolysis of corn cobs. The addition of 0.015 mg/mL EXPA17 resulted in a 14.00 % increase in glucose yield. Fourier-transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) analysis revealed that EXPA17 proteins disrupted hydrogen bonds in the amorphous regions of corn cobs, leading to a more porous and looser structure, thereby enhancing cellulose accessibility. Our work leveraged the synergistic effect between Expansin and lignocellulose from the same source of corn, providing a novel strategy to improve the efficiency of enzymatic hydrolysis of corn lignocellulose while potentially reducing the associated costs.

Saccharomyces cerevisiae for lignocellulosic ethanol production: a look at key attributes and genome shuffling.

These days, bioethanol research is looking at using non-edible plant materials, called lignocellulosic feedstocks, because they are cheap, plentiful, and renewable. However, these materials are complex and require pretreatment to release fermentable sugars. Saccharomyces cerevisiae, the industrial workhorse for bioethanol production, thrives in sugary environments and can handle high levels of ethanol. However, during lignocellulose fermentation, S. cerevisiae faces challenges like high sugar and ethanol concentrations, elevated temperatures, and even some toxic substances present in the pretreated feedstocks. Also, S. cerevisiae struggles to efficiently convert all the sugars (hexose and pentose) present in lignocellulosic hydrolysates. That's why scientists are exploring the natural variations within Saccharomyces strains and even figuring out ways to improve them. This review highlights why Saccharomyces cerevisiae remains a crucial player for large-scale bioethanol production from lignocellulose and discusses the potential of genome shuffling to create even more efficient yeast strains.

Constructing of ultrahigh pH-responsive polycarboxylic copolymers for enhanced recycling cellulase from enzymatic hydrolyzate of corncob resides

The high cost of the enzymatic hydrolysis of lignocellulose is a major problem for the industrialization of lignocellulose-based sugar platform technology. Cellulase recovery is a vital approach to reduce the enzymatic hydrolysis cost. In this study, ultrahigh pH-responsive copolymers (PAMAs) were obtained from methacrylamide, sodium methacrylate and [2-(methacryloyloxy)ethyl]-trimethylammonium chloride. A PAMA addition method coupled with substrate reabsorption was used to efficiently recover cellulase in a narrow pH range. The enzymatic hydrolysis system of corncob resides (pH = 5.0) was supplemented with 1.0 g/L PAMA-1. After hydrolysis, the PAMAs coprecipitate with cellulase when adjusting the pH value of the hydrolysate to 4.4. When coupled with substrate reabsorption, 50 % of the cellulase was recovered. This method therefore represents a feasible solution to reduce the high enzymatic hydrolysis cost of the industrialization of lignocellulose-based sugar platform technology.

Relation between structural feature and non-ionic surfactant improving enzymatic hydrolysis of lignocellulose

Relation between structural feature and non-ionic surfactant improving enzymatic hydrolysis of lignocellulose

S. cerevisiae ERG5DeltaNTH1DeltaAMS1Delta Construction Enhancing Stress Tolerance for Ethanol Production Increase in the Presence of Inhibitors and Mechanism Analysis based on the Comparative Transcriptomics

Saccharomyces cerevisiae is considered the most promising large-scale production strain with ethanol as the main product. The fermentation of Saccharomyces cerevisiae is generally inhibited under various stress conditions. Various inhibitors in the hydrolysate severely inhibit yeast proliferation and yeast accumulation. In this study, S. cerevisiae ERG5, NTH1, and AMS1 were knocked out to improve the yeast stress tolerance by the Clustered Regularly Interspaced Short Palindromic Repeats Cas9 (CRISPR-Cas9) technology. The result indicated that the stress tolerance of S. cerevisiae ERG5ΔNTH1ΔAMS1Δ mutant (S. cerevisiae SCENA) was remarkably improved compared with the wild-type strain. The contents of fecosterol, trehalose, and mannan in S. cerevisiae SCENA were 1.67, 1.53, and 1.47 folds compared with those in the control. The ethanol concentration in S. cerevisiae SCENA reached 16.5 g/L, which was 1.23 folds compared with the control using rice bran hydrolysate. Further, the transcriptome analysis indicated down-regulated differential expression genes (DEGs) in S. cerevisiae SCENA were mainly from cellular response to glucose, cell periphery, and plasma membrane. Up-regulated DEGs were mainly from spore wall assembly, fungal-type cell wall assembly, and ascospore wall assembly. Thus, S. cerevisiae SCENA could effectively produce ethanol using the fermentation of lignocellulosic hydrolysate in the presence of inhibitors by regulating fecosterol, trehalose, and mannan metabolisms.

Multilevel systemic engineering of Bacillus licheniformis for efficient production of acetoin from lignocellulosic hydrolysates

Bio-refining lignocellulosic resource offers a renewable and sustainable approach for producing biofuels and biochemicals. However, the conversion efficiency of lignocellulosic resource is still challenging due to the intrinsic inefficiency in co-utilization of xylose and glucose. In this study, the industrial bacterium Bacillus licheniformis was engineered for biorefining lignocellulosic resource to produce acetoin. First, adaptive evolution was conducted to improve acetoin tolerance, leading to a 19.6 % increase in acetoin production. Then, ARTP mutagenesis and 60Co-γ irradiation was carried out to enhance the production of acetoin, obtaining 73.0 g/L acetoin from glucose. Further, xylose uptake and xylose utilization pathway were rewired to facilitate the co-utilization of xylose and glucose, enabling the production of 60.6 g/L acetoin from glucose and xylose mixtures. Finally, this efficient cell factory was utilized for acetoin production from lignocellulosic hydrolysates with the highest titer of 68.3 g/L in fed-batch fermentation. This strategy described here holds great applied potential in the biorefinery of lignocellulose for the efficient synthesis of high-value chemicals.

Improvement of Saccharomyces cerevisiae strain tolerance to vanillin through heavy ion radiation combined with adaptive laboratory evolution

Vanillin is an inhibitor of lignocellulose hydrolysate, which can reduce the ability of Saccharomyces cerevisiae to utilize lignocellulose, which is an important factor limiting the development of the ethanol fermentation industry. In this study, mutants of vanillin-tolerant yeast named H6, H7, X3, and X8 were bred by heavy ion irradiation (HIR) combined with adaptive laboratory evolution (ALE). Phenotypic tests revealed that the mutants outperformed the original strain WT in tolerance, growth rate, genetic stability and fermentation ability. At 1.6 g/L vanillin concentration, the average OD600 value obtained for mutant strains was 0.95 and thus about 3.4-fold higher than for the wild-type. When the concentration of vanillin was 2.0 g/L, the glucose utilization rate of the mutant was 86.3 % within 96 h, while that of the original strain was only 70.0 %. At this concentration of vanillin, the mitochondrial membrane potential of the mutant strain recovered faster than that of the original strain, and the ROS scavenging ability was stronger. We analyzed the whole transcriptome sequencing map and the whole genome resequencing of the mutant, and found that DEGs such as FLO9, GRC3, PSP2 and SWF1, which have large differential expression multiples and obvious mutation characteristics, play an important role in cell flocculation, rDNA transcription, inhibition of DNA polymerase mutation and protein palmitoylation. These functions can help cells resist vanillin stress. The results show that combining HIR with ALE is an effective mutagenesis strategy. This approach can efficiently obtain Saccharomyces cerevisiae mutants with improved vanillin tolerance, and provide reference for obtaining robust yeast strains with lignocellulose inhibitor tolerance.

Advances in synthesis of succinic acid using yeast cell factories

Succinic acid is an important C4 platform compound that serves as a raw material for the production of 1,4-butanediol, tetrahydrofuran, and biodegradable plastics such as polybutylene succinate (PBS). Compared to the traditional petrochemical-based route that uses maleic anhydride as a raw material, the microbial fermentation method for producing succinic acid offers more sustainable economic value and environmental friendliness. Yeasts with good acid tolerance can achieve low-pH fermentation of succinic acid, significantly reducing the cost of product extraction. Therefore, constructing high-yield succinic acid yeast strains through metabolic engineering has garnered increasing attention. This paper systematically introduced the application value and market size of succinic acid, summarized the pathways and key enzymes involved in succinic acid synthesis in microorganisms, and elaborated on the latest research progress in the synthesis of succinic acid using yeast cell factories. It also presented the current status of succinic acid synthesis using non-food raw materials such as glycerol, acetic acid, lignocellulosic hydrolysate, and others as substrates by engineered yeast strains. Finally, the paper provided a prospect for low-pH succinic acid biomanufacturing based on yeast cell factories.