High throughput automated colorimetric method for the screening of l-lactic acid producing microorganisms

• Main Authors: Nadège Liaud, David Navarro, Nicolas Vidal, Jean-Claude Sigoillot, Sana Raouche
• Format: Article
• Language: English
• Published: Elsevier 2014-01-01
• Series: MethodsX
• Subjects: Automated screening for lactic acid production
• Online Access: http://www.sciencedirect.com/science/article/pii/S2215016114000156

Lactic acid is a valuable and fully degradable organic acid with promising applications in poly-lactic acid production (Taskila S and Ojamo, 2013 [1]). Despite their efficiency, the cost of the current lactic acid bio-processes is still an obstacle to this application (Miller et al., 2011 [2]). To ameliorate lactic acid producing strains, researchers are using mutations and metabolic engineering techniques, as well as medium optimization. All these studies necessitate a good and high throughput screening method. Currently, researchers mostly use HPLC methods which often necessitate sample preparation, are not stereospecific and do not allow high throughput. To help optimizing l-lactic acid production, we developed a high throughput colorimetric method inspired by the blood l-lactic acid detection method used for diagnosis (Lin et al., 1999 [3]). • Two sequential enzymatic reactions using l-lactate oxidase, peroxidase and ABTS (2,2′-azino-di-[3-ethylbenzthiazoine-sulfonate]), a chromogenic peroxidase substrate, are used to quantify l-lactate between 13.8 and 90 mg/l. • The accuracy of the method was ascertained before automation. • The method was successfully applied for the direct determination of l-lactate content in fungal culture supernatants.