A thermophilic L-lactic acid producer of high optical purity: isolation and identification

Abstract

Biodegradable polymers, specifically polylactide, are an important part of food packaging and medical devices. Microbiological synthesis uses cheap renewable raw materials and industrial waste to produce a high yield of lactic acid, the monomer of polylactide. This method needs new effective lactic acid producing strains, e.g., thermophilic bacteria. The research involved thermophilic bacterial strains isolated from soil and compost samples. Their ability to produce organic acids and extracellular enzymes was tested using the method of high-performance liquid chromatography (HPLC) and microbiological tests respectively. The real-time polymerase chain reaction method (PCR) detected L-lactate dehydrogenase structural genes of L-lactate dehydrogenase of Bacillaceae. Strain T7.1 was fermented using glucose and yeast extract as carbon and nitrogen sources, respectively. The optical purity of lactic acid was evaluated using quantitative gas chromatography on a chiral column to separate lactate isomers. The molecular genetic analysis of the 16S rRNA gene sequence was applied to identify strain T7.1. The chromatographic analysis proved that 10 out of 13 isolated thermophilic strains were effective lactic acid producers. They demonstrated proteolytic, amylolytic, or cellulase activities. During the fermentation, strain T7.1 produced 81 g/L of lactic acid with a peak productivity at 1.58 g/(L·h). The optical purity of the product exceeded 99.9% L-lactate. The genetic analysis identified strain T7.1 as Weizmannia coagulans (Bacillus coagulans). The research revealed a promising thermophilic producer of optically pure L-lactic acid. Further research is needed to optimize the cultivation conditions, design an effective and cheap nutrient medium, and develop engineering and technological solutions to increase the yield.

Keywords

• thermophilic bacteria
• bacillus
• weizmannia coagulans
• lactic acid
• l-lactate
• polylactide