Fermentative production of organic acids

Our offer

The Industrial Biotechnology Group of Fraunhofer IGB develops processes for the production of platform chemicals using renewable raw materials. The focus is on the establishment, optimization and scaling of bioconversion processes in which biogenic raw materials are converted into basic chemicals either by microorganisms (bacteria, yeast or fungal cultures) or enzymes. Fungal systems have proven to be particularly advantageous for the microbial production of malic acid and itaconic acid.

Spectrum of services

 

  • Selection and optimization of biocatalysts
  • Development of suitable conversion processes on a laboratory scale
  • Transfer of the conversion process to the pilot plant to m³ scale

Within process development, we look at all relevant parameters such as temperature, pO2, OUR, CER and RQ or composition of the medium and establish perfectly adjusted mineral salt media and feeding strategies (repeated-batch, fed-batch or continuous culture management). On the basis of a statistical evaluation of all process parameters, we first transfer the optimal process from the shake flask to the fermenter (scale-over) and then into technical and pilot scale (scale-up). In addition, the downstream processing of products is enhanced.

 

Malic acid made of xylose – fermentation at 1 m³ scale for the first time

Aspergillus oryzae
Microscopic picture of the fungus Aspergillus oryzae.

To date, malic acid has been used primarily in the food and beverage industry. It improves the shelf-life of baked products and provides the sour taste of jams and juices. But it also boasts considerable potential as a building block in the chemical industry. Together with succinic and fumaric acid, it belongs to the group of C4 dicarboxylic acids. C4 acids can be converted into 1,4-butandiol (BDO) – an important precursor for further conversion into a wide variety of chemicals, including plastics, polymers and resins; the possible applications for these chemicals range from golf balls to printing inks and cleaning agents.

Fermentative production of malic acid was developed through the collaboration of the Industrial Biotechnology working group at Fraunhofer IGB and the Biotechnological Processes group at Fraunhofer CBP. Fermentation was carried out with the fungus Aspergillus oryzae, which is designated as a harmless food additive according to the GRAS (generally recognized as safe) status of the US Food and Drug Administration (FDA). In addition to glucose, the strain can also utilize the C5 sugar xylose, which is the main component of hemicellulose and thus can be sourced from wood residues.

Initially, the process was optimized at the laboratory scale; it was then established in stirred reactors and finally successfully scaled up to the 1 m³ scale using the substrate xylose for the first time. Downstream processing could be demonstrated using crystallization. In doing so, several kilograms of malic acid were produced that are now available as a sample for application tests.

High concentrations of xylonic acid through process optimization

xylonic acid
Production of xylonic acid from xylose using Gluconobacter.

Xylonic acid as a replacement for gluconic acid

Gluconic acid is an important constituent of foodstuffs, construction materials and dyes [1]. The acid is produced from glucose, which is obtained from plants rich in starch and thus competes with the production of foodstuffs. An alternative to gluconic acid is xylonic acid: on the one hand, this has similar properties and, on the other hand, it can be obtained from plant components containing lignocellulose or from agricultural waste material. The aim was therefore to develop an efficient process for obtaining xylonic acid from xylose.

250 g/L xylonic acid through optimization

The fermentation-based conversion of xylose is conducted using whole cell catalysis (Gluconobacter sp.), with addition of oxygen as a second reactant. In contrast to competing solutions, fermentation with Gluconobacter sp. has the advantage of being a specific, sustainable and efficient conversion. To date, the team of Industrial Biotechnology Group has achieved a xylonic acid concentration of over 250 g/L through optimization – with a yield of over 90 percent. In the subsequent rudimentary purification process, xylonic acid was obtained at a purity of over 80 percent, which is adequate for technical applications.

Scale-up and sample quantities for application-related investigations

The scalability of the process has already been demonstrated at the Fraunhofer Center for Chemical-Biotechnological Processes CBP by the team in the Biotechnological Processes Group with the 100-liter fermentation, a scale-up to 300 liters is planned. We are already making smaller quantities available for investigations for specific applications. For example, xylonic acid can be tested as a substitute for gluconic acid as a curing retardant for concrete or chelating agent.

Literature

[1] Toivari, M.H., Y. Nygard, M. Penttila, L. Ruohonen, and M.G. Wiebe, Microbial D-xylonate production. Applied Microbiology and Biotechnology, 2012. 96(1): p. 1-8.

Itaconic acid

Microscope image of the fungus Aspergillus terreus.

The fermentative production process of itaconic acid dates back to 1932, at that time using Aspergillus itaconicus. Now, the Aspergillus terreus strain is primarily used. We were able to yield 137 g/L itaconic acid using A. terreus for biotechnological applications.

Furan dicarboxylic acid

During the development of a furan dicarboxylic acid (FDCA) production process, we were able to successfully establish whole cell catalysis with Pseudomonas putida by adding hydroxymethylfurfural from biomass containing lignocellulose. In the laboratory, we were able to achieve a yield of more than 80 percent at a concentration up to 20 g/L FDCA through precise reaction control. The subsequent scale-up was carried out in model experiments with scalable reactors or fermenters in the laboratory. The dimensionless key figures selected in these experiments allowed us to adapt the processes to the larger scale of a pilot plant.

Reference projects

CarboSurf – New processes for the fermentative production of glycolipid biosurfactants and sialylated carbohydrates

Duration: August 2015July 2018

CarboSurf is a project of the Bio-Based Industries Joint Undertakings (BBI JU). With 6 million Euro budget ,this project aims to develop new processes and solve bottlenecks in the fermentative production of biosurfactants and special carbohydrates.

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KomBiChemPro – Fine and platform chemicals from wood through combined chemical-biotechnological processes

Duration: November 2015 – May 2018

The joint KomBiChemPro project aims to combine various development work on the material use of biomass containing lignocellulose in an integrated biorefinery concept. The most important aspect is the manufacture of marketable products through robust processes and efficient process management. The basis of the project is the fractionation of lignocellulose using the organosolv process, which enables the three main components of lignocellulose (cellulose, hemicellulose, and lignin) to be obtained. The Industrial Biotechnology Group develops fermentation processes for malic and xylonic acid.

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BioQED – Quod Erat Demonstrandum: Large scale demonstration for the bio-based bulk chemicals BDO and IA aiming at cost reduction and improved sustainability

Duration: January 2014 – December 2017

In the EU project Bio-QED, Fraunhofer IGB and its branch institute Fraunhofer CBP are conducting research into fermentative production processes to produce basic chemicals and regarding the scale-up of the processes. The Institute is concentrating in particular on the itaconic acid value chain. In 2017, Fraunhofer IGB and CBP held a demonstration workshop on this topic in which project highlights and results were presented.

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BioConSepT – Integration of Bio-Conversion and Separation Technology for the production and application of platform chemicals from 2nd generation biomass

Duration: January 2012 – December 2015

BioConSepT focuses on the material use of so-called second generation raw materials such as lignocellulose or oils and fats that cannot be used for food production. Both enzymatic, microbial and chemical reactions are used and, if necessary, combined in the production chains. The introduction of continuous processes, new reactors and selective separation technologies should achieve breakthroughs in cost reduction and sustainability of these processes.