Enantioselective lactic acid production by an Enterococcus faecium strain showing potential in agro-industrial waste bioconversion: physiological and proteomic studies

J Biotechnol. 2014 Mar 10:173:31-40. doi: 10.1016/j.jbiotec.2014.01.014. Epub 2014 Jan 19.

Abstract

The growing demand of biodegradable plastic polymers is increasing the industrial need of enantiospecific l-lactic acid (l-LA), the building block to produce polylactides. The most suitable industrial strategy to obtain high amounts of LA is the microbial fermentation of fruit and vegetable wastes by lactic acid bacteria (LAB). In this paper seven LAB strains from our laboratory collection, were screened for their ability to produce the highest amount of pure l-LA. A strain of Enterococcus faecium (LLAA-1) was selected and retained for further investigations. E. faecium LLAA-1 was grown in different culture media supplemented with the most abundant sugars present in agricultural wastes (i.e., glucose, fructose, cellobiose and xylose) and its ability to metabolize them to l-LA was evaluated. All tested sugars proved to be good carbon sources for the selected strain, except for xylose, which resulted in unsatisfactory biomass and LA production. Growth under aerobic conditions further stimulated l-LA production in fructose supplemented cultures with respect to anoxic-grown cultures. Proteomic profiles of E. faecium LLAA-1 grown in aerobiosis and anoxia were compared by means of two-dimensional electrophoresis followed by MALDI-TOF mass spectrometry. Seventeen proteins belonging to three main functional groups were differentially expressed: the biosynthesis of 6 proteins was up-regulated in aerobic-grown cultures while 11 proteins were biosynthesized in higher amounts in anoxia. The de novo biosynthesis of the f-subunit of alkyl hydroperoxide reductase involved in the re-oxidation of NADH seems the key element of the global re-arrangement of E. faecium LLAA-1 metabolism under aerobic conditions. An improved oxidative catabolism of proteinaceous substrates (i.e., protein hydrolisates) seems the main phenomenon allowing both higher biomass growth and improved LA production under these conditions.

Keywords: Fructose; Lactic acid bacteria; Oxygen; Proteomics; l-Lactic acid.

MeSH terms

  • Aerobiosis
  • Bacterial Proteins / analysis*
  • Biomass
  • Cellobiose / metabolism
  • Culture Media / chemistry*
  • Enterococcus faecium / classification
  • Enterococcus faecium / metabolism*
  • Fermentation
  • Fructose / metabolism
  • Gene Expression Regulation, Bacterial
  • Glucose / metabolism
  • Industrial Waste
  • Lactic Acid / metabolism*
  • Peroxiredoxins / metabolism
  • Proteomics
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Xylose / metabolism

Substances

  • Bacterial Proteins
  • Culture Media
  • Industrial Waste
  • Cellobiose
  • Fructose
  • Lactic Acid
  • Xylose
  • Peroxiredoxins
  • Glucose