Elsevier

Process Biochemistry

Volume 41, Issue 6, June 2006, Pages 1461-1465
Process Biochemistry

Short communication
Effective purification of succinic acid from fermentation broth produced by Mannheimia succiniciproducens

https://doi.org/10.1016/j.procbio.2006.01.020Get rights and content

Abstract

The present study deals with the development of purification and separation processes required to produce the highly purified succinic acid from the fermentation broth produced by recombinant microorganism, Mannheimia succiniciproducens. The newly developed process consists of the pretreatment process such as reactive extraction and vacuum distillation step and the crystallization process for the highly purified succinic acid production. By-produced acids were effectively removed by the reactive extraction as a primary separation. In addition, the crystallization was applied without adding any salts to produce highly purified succinic acid. The purified succinic acid, with 99.8% purity and 73.1% yield rate was obtained through this newly developed purification process.

Introduction

Succinic acid is a common metabolite formed by plants, animals and microorganisms. It is a C4 dicarboxylic acid produced as an intermediate of the tricarboxylic acid cycle (TCA), and also as one of the fermentation products of energy metabolism [1], [2]. As the importance of succinic acid for use as a biodegradable polymer has increased, the biological production by fermentation has been focused on as the alternative to the petrochemical-based process [2], [3]. Newly discovered facultative anaerobic bacteria Actinobacilllus succinogenes and Mannheimia succiniciproducens are considered as the effective succinic acid producers because they can endure high glucose osmotic pressure and produce significant amounts of succinic acid with a high productivity [4], [5]. The fermentation broth used in this study was cultured by M. succiniciproducens, which was isolated from bovine rumens by Lee et al., and the yield was 22.3 g succinic acid/l. Contaminated organic acids were also produced as a by-product with a gram ratio of succinic acid to by-produced acids of 1.76 [4], [5], [6], [7]. Contaminated organic acids adversely affect the recovery of succinic acid as well as its yield in the fermentor [3], [4], [5].

Therefore, in the production of succinic acid by fermentation, the separation process for succinic acid should be required due to the many impurities in the fermentation broth, such as by-produced acids, carbon sources and salts [4], [8]. Normally, the downstream purification cost accounts for 60–70% of the production cost in the fermentation based process [9]. For the economical recovery process of succinic acid from the fermentation broth, several possible alternatives such as precipitation, distillation, electrodialysis and extraction have been reported [10], [11], [12], [13]. However, it is impossible to apply only one separation process for product recovery, concentration, acidification and purification of succinic acid to its required purity. The traditional succinic acid recovery method is based on precipitation and crystallization technology, acidification by ion-exchange resins and crystallization process [8], [14]. However, the production of succinic acid by this process is very costly and it requires a much more complex process for commercial use.

Therefore, in the present study, we propose an efficient process for the development of the more effective separation and production of succinic acid. Throughout our process, reactive extraction has been proposed to as a primary separation method for removal of by-produced acids from the fermentation broth [15], [16], [17]. Then, pretreated samples by reactive extraction were further purified by the crystallization that was conducted at a different pH. It was the purpose of this investigation to purify succinic acid from the fermentation broth, and to establish a simpler and more effective complex purification process. We obtained highly purified succinic acid with the pretreatment of reactive extractions and followed by crystallization.

Section snippets

Organic chemicals

Tri-n-octylamine (TOA; Junsei, Japan, 99%) was used as the extractant without further purification. Diluent was used in the reactive extraction of succinic acid because of the high viscosity and the corrosive property of TOA. 1-Octanol (Aldrich, 99.9%) was used as diluent in this work. The concentration of TOA in the organic phase was 0.25 mol/kg as the basis of 1-octanol. The pH of the fermentation broth was adjusted by adding either NaOH or HCl solution. The pH of the fermentation broth was

Pretreatment of fermentation broth by reactive extraction

In this study, the effective purification process of succinic acid was investigated from fermentation broth produced by M. succiniciproducens. To be cost effective, the separation process required the removal of cells and protein-like impurities, conversion of the succinic salt into free acid, and polishing of the free acid to its required purity. Thus, the complex separation technology is applied to simultaneous reactive extraction, vacuum distillation and crystallization for the effective

Acknowledgements

This work was supported by the Genome-based Integrated Bioprocess Project of the Ministry of Science and Technology. Further supports by Brain Korea 21 (BK21).

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