A novel fermentation strategy for removing the key inhibitor acetic acid and efficiently utilizing the mixed sugars from lignocellulosic hydrolysates

Gluconic acid is a non-volatile acid that has many applications in food, pharmaceutical and cleaning fields. Gluconic acid has been detected as main oxidation product of Acetobacter and Gluconobacter strains growing on grape must, and it plays an important role in Traditional Balsamic Vinegar. Commonly, high gluconate vinegars have a greater physical stability and a greater preference by consumers because are perceived less pungent. In fact, gluconic acid reduces the pH and increases fixed acidity of the vinegar without increasing the sensation of pungency typical of acetic acid. Its taste is acid but mild sweet and, therefore, gluconic acid has influence on the sensory complexity of the vinegar. The aim of this work is to set up a fermentation procedure that improves the quality of balsamic vinegar by using selected yeasts and acetic acid bacteria strains able to oxidize glucose in grape must-based media having a different sugars concentration. In particular, Saccharomycodes ludwigii UMCC 297 and Acetobacter pasteurianus UMCC 1754 strains were chosen as selected starter cultures for small-scale fermentation of cooked grape must, to evaluate the physical-chemical parameters affecting gluconic acid production in the obtained vinegar. The strains used and the control of all production process have been fundamental for obtaining the vinegar with the desired characteristics.

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Elimination of Glycerol Production in Anaerobic Cultures of a Saccharomyces cerevisiae Strain Engineered To Use Acetic Acid as an Electron Acceptor

Applied and Environmental Microbiology ◽

10.1128/aem.01772-09 ◽

2009 ◽

Vol 76 (1) ◽

pp. 190-195 ◽

Cited By ~ 107

Author(s):

Víctor Guadalupe Medina ◽

Marinka J. H. Almering ◽

Antonius J. A. van Maris ◽

Jack T. Pronk

Keyword(s):

Saccharomyces Cerevisiae ◽

Acetic Acid ◽

Coenzyme A ◽

Anaerobic Growth ◽

Industrial Biotechnology ◽

Glycerol Production ◽

Complete Replacement ◽

Lignocellulosic Hydrolysates ◽

Ethanol Yields ◽

Acetate Reduction

ABSTRACT In anaerobic cultures of wild-type Saccharomyces cerevisiae, glycerol production is essential to reoxidize NADH produced in biosynthetic processes. Consequently, glycerol is a major by-product during anaerobic production of ethanol by S. cerevisiae, the single largest fermentation process in industrial biotechnology. The present study investigates the possibility of completely eliminating glycerol production by engineering S. cerevisiae such that it can reoxidize NADH by the reduction of acetic acid to ethanol via NADH-dependent reactions. Acetic acid is available at significant amounts in lignocellulosic hydrolysates of agricultural residues. Consistent with earlier studies, deletion of the two genes encoding NAD-dependent glycerol-3-phosphate dehydrogenase (GPD1 and GPD2) led to elimination of glycerol production and an inability to grow anaerobically. However, when the E. coli mhpF gene, encoding the acetylating NAD-dependent acetaldehyde dehydrogenase (EC 1.2.1.10; acetaldehyde + NAD+ + coenzyme A ↔ acetyl coenzyme A + NADH + H+), was expressed in the gpd1Δ gpd2Δ strain, anaerobic growth was restored by supplementation with 2.0 g liter−1 acetic acid. The stoichiometry of acetate consumption and growth was consistent with the complete replacement of glycerol formation by acetate reduction to ethanol as the mechanism for NADH reoxidation. This study provides a proof of principle for the potential of this metabolic engineering strategy to improve ethanol yields, eliminate glycerol production, and partially convert acetate, which is a well-known inhibitor of yeast performance in lignocellulosic hydrolysates, to ethanol. Further research should address the kinetic aspects of acetate reduction and the effect of the elimination of glycerol production on cellular robustness (e.g., osmotolerance).

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Ectodesmata as Revealed By Freeze-Etch Preparations of Plant Epidermal Cell Walls

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100072782 ◽

1973 ◽

Vol 31 ◽

pp. 468-469

Author(s):

N.C. Lyon ◽

W. C. Mueller

Keyword(s):

Electron Microscopy ◽

Acetic Acid ◽

Nitric Acid ◽

Oxalic Acid ◽

Light Microscopy ◽

Epidermal Cell ◽

Cell Walls ◽

Plant Viruses ◽

Plant Leaves ◽

Thin Sections

Schumacher and Halbsguth first demonstrated ectodesmata as pores or channels in the epidermal cell walls in haustoria of Cuscuta odorata L. by light microscopy in tissues fixed in a sublimate fixative (30% ethyl alcohol, 30 ml:glacial acetic acid, 10 ml: 65% nitric acid, 1 ml: 40% formaldehyde, 5 ml: oxalic acid, 2 g: mecuric chloride to saturation 2-3 g). Other workers have published electron micrographs of structures transversing the outer epidermal cell in thin sections of plant leaves that have been interpreted as ectodesmata. Such structures are evident following treatment with Hg++ or Ag+ salts and are only rarely observed by electron microscopy. If ectodesmata exist without such treatment, and are not artefacts, they would afford natural pathways of entry for applied foliar solutions and plant viruses.

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