In Malolactic Fermentation and Bacteria

By Dr. Molly Kelly, Enology Extension Educator, Department of Food Science. 

As harvest comes to a close we have planned which wines will be going through malolactic fermentation (MLF). This article provides some information to assist you in dealing with a potentially difficult MLF.

Malolactic fermentation (MLF) is a process of chemical change in wine in which L-malic acid is converted to L-lactic acid and carbon dioxide. This process is normally conducted by lactic acid bacteria (LAB) including Oenococcus oeni, Lactobacillus spp. and Pediococcus spp. O.oeni is the organism typically used to conduct MLF due to its tolerance to low pH, high ethanol and SO2. Most commercial strains are designed to produce favorable flavor profiles.

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Although inoculation with a commercial starter is recommended, MLF may occur spontaneously. The lag phase associated with spontaneous MLF may increase the risk of spoilage organisms as well as the production of volatile acidity. Inoculation with a LAB culture can help avoid these problems by providing the cell population needed to successfully conduct MLF (more than 2×106 cells/mL). The compatibility of yeast and LAB should be taken into account since failed MLF may be due to incompatibility between these two organisms.

The key to a successful MLF is to manage the process and to monitor the progress. Although there has been extensive research on the MLF process, it may still be difficult to initiate at times. The possible causes of difficult MLF have been studied less extensively than those of stuck/sluggish alcoholic fermentation. In this article, factors that may influence the start and successful completion of MLF will be discussed.

The main chemical properties that influence MLF are well known: pH, temperature, ethanol and SO2 concentration. A study by Vaillant et al (1995) investigating the effects of 11 physico-chemical parameters, identified ethanol, pH and SO2 as having the greatest inhibitory effect on the growth of LAB in wine.


Generally, LAB prefer increased pH’s and usually, minimal growth occurs at pH 3.0. Under winemaking conditions, pH’s above 3.2 are advised. The pH will determine the dominant species of LAB in the must or wine.  At a low pH (3.2 to 3.4) O. oeni is the most abundant LAB species, while at higher pH (3.5 to 4.0), Lactobacillus and Pediococcus will out-number Oenococcus.


MLF is generally inhibited by low temperatures. Research demonstrates that MLF occurs faster at temperatures of 200 C (68˚F) and above versus 150C (59˚F) and below. In the absence of SO2 the optimum temperature range for MLF is 23-250C (73.4˚F-77˚F) with maximum malic acid conversion taking place at 20-250C (68˚F-77˚F). However, with increasing SO2 levels, these temperatures decrease and 200C (68˚F) may be more acceptable.


LAB are ethanol-sensitive with slow or no growth occurring at approximately 13.5%. Commercial O. oeni strains are preferred starter cultures due to tolerance to ethanol.  The fatty acid composition of the cell membrane of LAB can be impacted by ethanol content.

Sulfur dioxide

LAB may be inhibited by the SO2 produced by yeast during alcoholic fermentation. A total SO2 concentration of more than 50 ppm generally limits LAB growth, especially at lower pH where a larger portion of SO2 is in the antimicrobial form. Generally, it is not recommended to add SO2 after alcoholic fermentation if MLF is desired.

Some of the lesser known factors impacting MLF are discussed below.

Fatty Acids

MLF can be inhibited by medium chain fatty acids (octanoic and decanoic acids) produced by yeast. It is difficult to finish MLF when octanoic acid content is over 25 mg/L and/or decanoic acid is over 5 mg/L. Bacterial strains that tolerate high concentrations of octanoic and decanoic acids may be important in successful MLF. It is important to check your supplier regarding strain specifications. Yeast hulls may be added before the bacteria are inoculated (0.2g/L) to bind fatty acids. Yeast hulls may also supply unsaturated fatty acids, amino acids and assist with CO2 release.


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