L-Lactic acid production from glycerol coupled with acetic acid metabolism by Enterococcus faecalis without carbon loss.

Glycerol is a by-product in the biodiesel production process and considered as one of the prospective carbon sources for microbial fermentation including lactic acid fermentation, which has received considerable interest due to its potential application. Enterococcus faecalis isolated in our laboratory produced optically pure L-lactic acid from glycerol in the presence of acetic acid. Gas chromatography-mass spectrometry analysis using [1, 2-(13)C2] acetic acid proved that the E. faecalis strain QU 11 was capable of converting acetic acid to ethanol during lactic acid fermentation of glycerol. This indicated that strain QU 11 restored the redox balance by oxidizing excess NADH though acetic acid metabolism, during ethanol production, which resulted in lactic acid production from glycerol. The effects of pH control and substrate concentration on lactic acid fermentation were also investigated. Glycerol and acetic acid concentrations of 30 g/L and 10 g/L, respectively, were expected to be appropriate for lactic acid fermentation of glycerol by strain QU 11 at a pH of 6.5. Furthermore, fed-batch fermentation with 30 g/L glycerol and 10 g/L acetic acid wholly exhibited the best performance including lactic acid production (55.3 g/L), lactic acid yield (0.991 mol-lactic acid/mol-glycerol), total yield [1.08 mol-(lactic acid and ethanol)]/mol-(glycerol and acetic acid)], and total carbon yield [1.06 C-mol-(lactic acid and ethanol)/C-mol-(glycerol and acetic acid)] of lactic acid and ethanol. In summary, the strain QU 11 successfully produced lactic acid from glycerol with acetic acid metabolism, and an efficient fermentation system was established without carbon loss.

[1]  K. Rumbold,et al.  Microbial utilization of crude glycerol for the production of value-added products , 2012, Journal of Industrial Microbiology & Biotechnology.

[2]  A. Bizzini,et al.  Glycerol Is Metabolized in a Complex and Strain-Dependent Manner in Enterococcus faecalis , 2009, Journal of bacteriology.

[3]  Y. Tashiro,et al.  Isolation and characterisation of lactic acid bacterium for effective fermentation of cellobiose into optically pure homo l-(+)-lactic acid , 2011, Applied Microbiology and Biotechnology.

[4]  James M Clomburg,et al.  Escherichia coli Strains Engineered for Homofermentative Production of d-Lactic Acid from Glycerol , 2010, Applied and Environmental Microbiology.

[5]  Y. Tashiro,et al.  Lactic acid production from lignocellulose-derived sugars using lactic acid bacteria: overview and limits. , 2011, Journal of biotechnology.

[6]  M. Xian,et al.  Production of optically pure d-lactate from glycerol by engineered Klebsiella pneumoniae strain. , 2014, Bioresource technology.

[7]  A. Blanch,et al.  Identification of Enterococcus spp. with a Biochemical Key , 1999, Applied and Environmental Microbiology.

[8]  T. Peppard,et al.  Solid-Phase Microextraction for Flavor Analysis , 1994 .

[9]  Yukihiro Tashiro,et al.  Recent advances to improve fermentative butanol production: genetic engineering and fermentation technology. , 2015, Journal of bioscience and bioengineering.

[10]  Yuki Doi,et al.  Pyruvate Formate-Lyase Is Essential for Fumarate-Independent Anaerobic Glycerol Utilization in the Enterococcus faecalis Strain W11 , 2014, Journal of bacteriology.

[11]  Y. Tashiro,et al.  Continuous d-lactic acid production by a novelthermotolerant Lactobacillus delbrueckii subsp. lactis QU 41 , 2011, Applied Microbiology and Biotechnology.

[12]  Hong Liu,et al.  Microbial Conversion of Waste Glycerol from Biodiesel Production into Value-Added Products , 2013 .

[13]  P. Cunningham,et al.  Anaerobic regulation of the adhE gene, encoding the fermentative alcohol dehydrogenase of Escherichia coli , 1993, Journal of bacteriology.

[14]  J. Deutscher,et al.  Aerobic glycerol dissimilation via the Enterococcus faecalis DhaK pathway depends on NADH oxidase and a phosphotransfer reaction from PEP to DhaK via EIIADha. , 2012, Microbiology.

[15]  Wei-guo Zhang,et al.  Improvement of l-lactic acid production by osmotic-tolerant mutant of Lactobacillus casei at high temperature , 2010, Applied Microbiology and Biotechnology.

[16]  Y. Tashiro,et al.  Recent advances in lactic acid production by microbial fermentation processes. , 2013, Biotechnology advances.

[17]  D. Clark,et al.  Role of NAD in regulating the adhE gene of Escherichia coli , 1996, Journal of bacteriology.

[18]  Roberto Mazzoli,et al.  Towards lactic acid bacteria-based biorefineries. , 2014, Biotechnology advances.

[19]  James M Clomburg,et al.  Understanding and harnessing the microaerobic metabolism of glycerol in Escherichia coli , 2009, Biotechnology and bioengineering.

[20]  R. Gonzalez,et al.  Anaerobic fermentation of glycerol: a path to economic viability for the biofuels industry. , 2007, Current opinion in biotechnology.

[21]  Y. Tashiro,et al.  Fermentative production of lactic acid from renewable materials: recent achievements, prospects, and limits. , 2015, Journal of bioscience and bioengineering.

[22]  Zahid Anwar,et al.  Recent trends in lactic acid biotechnology: A brief review on production to purification , 2014 .

[23]  Ramon Gonzalez,et al.  Anaerobic fermentation of glycerol: a platform for renewable fuels and chemicals. , 2013, Trends in biotechnology.

[24]  G. Shi,et al.  High-efficiency conversion of glycerol to D-lactic acid with metabolically engineered Escherichia coli , 2012 .

[25]  Dehua Liu,et al.  Strain isolation and optimization of process parameters for bioconversion of glycerol to lactic acid , 2009 .

[26]  Y. Jang,et al.  Bio‐based production of C2–C6 platform chemicals , 2012, Biotechnology and bioengineering.

[27]  H. Chang,et al.  Characteristics and glycerol metabolism of fumarate-reducing Enterococcus faecalis RKY1. , 2001, Biotechnology and bioengineering.

[28]  O. Pop,et al.  L (+)-lactic acid production by pellet-form Rhizopus oryzae NRRL 395 on biodiesel crude glycerol , 2013, Microbial Cell Factories.

[29]  A. M. Strasser de Saad,et al.  Glycerol Metabolism of Lactobacillus rhamnosus ATCC 7469: Cloning and Expression of Two Glycerol Kinase Genes , 2004, Journal of Molecular Microbiology and Biotechnology.