Simultaneous Quantification of L-arginine and Monosaccharides during Fermentation: An Advanced Chromatography Approach.

Mireille Ginésy,Ulrika Rova,Daniela Rusanova-Naydenova,Josefine Enman

doi:10.3390/molecules24040802

Mireille Ginésy, Ulrika Rova + Show 2 more

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https://doi.org/10.3390/molecules24040802

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Abstract

Increasing demand for L-arginine by the food and pharmaceutical industries has sparked the search for sustainable ways of producing it. Microbial fermentation offers a suitable alternative; however, monitoring of arginine production and carbon source uptake during fermentation, requires simple and reliable quantitative methods compatible with the fermentation medium. Two methods for the simultaneous quantification of arginine and glucose or xylose are described here: high-performance anion-exchange chromatography coupled to integrated pulsed amperometric detection (HPAEC-IPAD) and reversed-phase ultra-high-performance liquid chromatography combined with charged aerosol detection (RP-UHPLC-CAD). Both were thoroughly validated in a lysogeny broth, a minimal medium, and a complex medium containing corn steep liquor. HPAEC-IPAD displayed an excellent specificity, accuracy, and precision for arginine, glucose, and xylose in minimal medium and lysogeny broth, whereas specificity and accuracy for arginine were somewhat lower in medium containing corn steep liquor. RP-UHPLC-CAD exhibited high accuracy and precision, and enabled successful monitoring of arginine and glucose or xylose in all media. The present study describes the first successful application of the above chromatographic methods for the determination and monitoring of L-arginine amounts during its fermentative production by a genetically modified Escherichia coli strain cultivated in various growth media.

Highlights

L-arginine is an important dietary supplement in animal feed [1,2,3] and has several potential clinical applications, such as in the treatment or prevention of cardiovascular diseases, diabetes type II, Alzheimer’s disease or infertility [4,5,6]
The tailored short HPAEC-IPAD method uses a detection waveform recommended for amino acid analysis (“Gold, pH/Ag/AgCl RE, AAA”) and the reference electrode in pH mode [40]
As the objective of this study was to establish the suitability of this method for fermentation samples, accuracy was tested in diluted M9’, lysogeny broth (LB), and corn steep liquor (CSL) media

Summary

Introduction

L-arginine (hereafter referred to as arginine) is an important dietary supplement in animal feed [1,2,3] and has several potential clinical applications, such as in the treatment or prevention of cardiovascular diseases, diabetes type II, Alzheimer’s disease or infertility [4,5,6]. An increased demand for arginine has driven the search for a more sustainable production that replaces keratin hydrolysis [11]. This has led to the use of Corynebacterium glutamicum and Corynebacterium crenatum for the fermentative production of arginine [11,12]. Unlike members of the Corynebacterium genus, Escherichia coli can metabolize five-carbon sugars, making it another potential candidate for arginine production, even if at lower yields [13]

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