A complete industrial system for economical succinic acid production by Actinobacillus succinogenes
Introduction
The development of biorefineries has recently attracted increasing attention as a means of providing sustainable alternative solutions to convert renewable agricultural materials into numerous economically viable products (fuels, chemicals, and other target molecules) and offer competitive performance compared to traditional petrochemical refineries (Ji et al., 2009, Dorado et al., 2009, Lynd and Wyman, 1999). Many chemicals that could only be produced by chemical processes in the past could potentially be generated biologically from annually renewable resources (Ragauskas et al., 2006).
Microbial production of succinic acid is a good example. Interest in this bioprocess has been increasing recently due to that succinic acid could serve as a precursor to various commodity chemicals used in industries like adipic acid, 1,4-butanediol, tetrahydrofuran, N-methyl pyrrolidinone, 2-pyrrolidinone, succinate salts, gamma-butyrolactone, poly-butyrate succinate, polyamides, and various green solvents (McKinlay et al., 2007, Zheng et al., 2009). Succinic acid is traditionally manufactured from petrochemicals through expensive processes. The biologic production of succinic acid from renewable resources and the greenhouse gas CO2 would alleviate the dependence on specialty chemical production from petroleum. However, bio-based succinic acid has not yet become competitive with petrochemical-based production, mainly because of high production costs (McKinlay et al., 2007, Song and Lee, 2006). There is therefore a need to develop a cost-effective process for succinic acid production from renewable resources.
Among succinic acid producers such as Actinobacillus succinogenes (Guettler et al., 1999), Anaerobiospirillum succiniciproducens (Lee et al., 2010), Mannheimia succiniciproducens (Lee et al., 2002), and recombinant Escherichia coli (Jantama et al., 2008), A. succinogenes not only could efficiently produce succinate (Mckinlay et al., 2007), but also could use a wide range of carbon sources, including lactose, xylose, arabinose, cellobiose, and other reduced sugars, to produce succinic acid as the major end product (Guettler et al., 1999, Van der Werf et al., 1997). The cost of carbon sources has been reduced by many renewable resources, such as straw (Zheng et al., 2009), corn fiber (Chen et al., 2010a), crop stalk wastes (Li et al., 2010c), whey (Wan et al., 2008), wheat (Dorado et al., 2009), and cane molasses (Liu et al., 2008b), all of which could be hydrolyzed into mixed sugars. The process of succinic acid production from lignocellulosic materials is economically attractive. In addition to carbon sources, nitrogen sources are also used to reduce the cost of succinic acid fermentation. Our previous research have shown that A. succinogenes NJ113 could use an enzymatic hydrolysate of spent yeast cells as a nitrogen source for succinic acid production in glucose-based and corn fiber hydrolysate-based media (Jiang et al., 2010, Chen et al., 2010a).
Succinic acid is an acid product. Large amounts of alkaline neutralizer are required to maintain the pH during succinic acid fermentation. The cost of alkaline neutralizer accounts for a significant portion of raw material costs. Majority of studies on succinic acid production have used MgCO3 as alkaline neutralizer to achieve high product concentrations. Nevertheless, the cost of MgCO3 supplementation is not practical for industrial succinic acid fermentation. In a previous study, our laboratory also investigated the replacement of MgCO3 with Na2CO3, NaHCO3, or NH3·H2O as alkaline neutralizer (Li et al., 2010a, Li et al., 2010b, Ye et al., 2010). However, this could only happen under low initial carbon source concentrations. There have been no reports regarding the use of mixed alkalis as alkaline neutralizer for regulating pH for succinic acid production.
In this study, mixed alkalis were first used as a novel method for regulating pH for succinic acid production. A complete industrial system using lignocellulose hydrolysate as the carbon source, waste yeast hydrolysate as the nitrogen source, and mixed alkali as the alkaline neutralizer to achieve high-yield, economical succinic acid production by A. succinogenes was developed.
Section snippets
Preparation of corn stover hydrolysate
Corn stover hydrolysate was from BBCA Group Corp. (Anhui, China). The corn stover was pretreated with 1% (w/v) diluted aqueous sulfuric acid solution, and then with 12% (w/v) 20 FPU g−1 cellulase with 0.05 M Tris–HCl as pH buffer. Corn stover hydrolysate components consisted of 326.80 g/L total sugar (of which glucose was 180.12 g/L, xylose was 92.20 g/L, and arabinose was 44.61 g/L), 1.35 g/L furfural, and 0.59 g/L hydroxymethylfurfural (HMF). Hydrolysate was diluted into different concentrations as
Utilization of corn stover hydrolysate as carbon source
Lignocellulosic materials represent the largest reservoir of potentially fermentable carbohydrates. These carbohydrates, obtained via either acid-promoted hydrolysis or by enzymatic hydrolysis, are intrinsically a mixture of pentoses, such as, xylose and arabinose, and hexoses, such as, glucose and mannose, among others (Stephanopoulos, 2007). Generally, complete hydrolysis of corn stover ideally generates a solution primarily containing glucose and xylose at an approximate ratio of 2:1 (w/w) (
Conclusion
In this study, economical succinic acid production through the fermentation of corn stover hydrolysate, biotin-supplemented YCH, and a Mg(OH)2 and NaOH mixture (1:1, w/w) by A. succinogenes NJ113 was developed. Using the cheap medium and optimally mixed alkaline neutralizers, a maximum succinic acid yield of 72.6% was achieved, which is comparable to the yield using glucose, YE, and MgCO3. From an industrial standpoint, the cheap carbon and nitrogen sources, as well as the mixed alkaline
Acknowledgements
This work was supported by the National Basic Research Program of China (Project No. 2009CB724701), the National Natural Science Foundation of China (Project No. 21076105), the State Key Laboratory of Materials-Oriented Chemical Engineering Foundation of Nanjing University of Technology, the Science and Technology Achievement Transformation Project of Jiangsu Province, and Qing Lan Project of Jiangsu Province.
References (34)
- et al.
Kinetic study of succinic acid production by Actinobacillus succinogenes ZT-130
Proc. Biochem.
(2008) - et al.
Cereal-based biorefinery development: utilisation of wheat milling by-products for the production of succinic acid
J. Biotechnol.
(2009) - et al.
Development of an industrial medium for economical 2,3-butanediol production through co-fermentation of glucose and xylose by Klebsiella oxytoca
Bioresour. Technol.
(2009) - et al.
Succinic acid production by Anaerobiospirillum succiniciproducens: effects of the H2/CO2 supply and glucose concentration
Enzyme Microbial. Technol.
(1999) - et al.
Enhanced production of succinic acid by Actinobacillus succinogenes with reductive carbon source
Proc. Biochem.
(2010) - et al.
Efficient conversion of crop stalk wastes into succinic acid production by Actinobacillus succinogenes
Bioresour. Technol.
(2010) - et al.
Economical succinic acid production from cane molasses by Actinobacillus succinogenes
Bioresour. Technol.
(2008) - et al.
Fermentation of lignocellulosic hydrolysates. I: inhibition and detoxification
Bioresour. Technol.
(2000) - et al.
Development of culture media containing spent yeast cells of Debaryomyces hansenii and corn steep liquor for lactic acid production with Lactobacillus rhamnosus
Int. J. Food Microbiol.
(2004) - et al.
Production of succinic acid by bacterial fermentation
Enzyme Microb. Technol.
(2006)
Dilute sulfuric acid cycle spray flow-through pretreatment of corn stover for enhancement of sugar recovery
Bioresour. Technol.
Fermentative production of succinic acid from straw hydrolysate by Actinobacillus succinogenes
Bioresour. Technol.
Effect of different carbon sources on the production of succinic acid using metabolically engineered Escherichia coli
Biotechnol. Prog.
Official Methods of Analysis Method 988.05
Comparative kinetic effects of Mn (II), Mg (II) and the ATP/ADP ratio on phosphoenolpyruvate carboxykinases from Anaerobiospirillum succiniciproducens and Saccharomyces cerevisiae
Protein J.
Succinic acid production from acid hydrolysate of corn fiber by Actinobacillus succinogenes
Appl. Biochem. Biotechnol.
Cited by (87)
Enzymatic hydrolysis of organosolv-pretreated corncob and succinic acid production by Actinobacillus succinogenes
2024, Industrial Crops and ProductsComparison of hydrothermolysis and mild-alkaline pretreatment methods on enhancing succinic acid production from hydrolyzed corn fiber
2024, Enzyme and Microbial TechnologyContinuous production of succinic acid from glycerol: A complete experimental and computational study
2023, Bioresource TechnologyEmpirical analysis of large-scale bio-succinic acid commercialization from a technoeconomic and innovation value chain perspective: BioAmber biorefinery case study in Canada
2021, Renewable and Sustainable Energy ReviewsOptimization of anaerobic fermentation of Actinobacillus succinogenes for increase the succinic acid production
2020, Biocatalysis and Agricultural BiotechnologyCitation Excerpt :This study encourages the use of renewable feedstocks and shows that a lower energy input, waste generation and carbon footprint can be employed to generate succinic acid. Future research should optimize the levels of NaHCO3 for succinic acid production, as carbonate is a crucial CO2 source for anaplerotic reactions and can act as a pH buffer (Li et al., 2011; Tan et al., 2018; Zou et al., 2011). Better control of pH and CO2 levels during fermentation is important to maximize succinic acid yield.