William McKeever

Food Science Paper

HN300

Fermented Dairy and the Role of Exopolysaccharides in Food Science

Traditions of fermentation have existed in most cultures since before the earliest recordings of history. Several forms of fermented dairy have been touted throughout history as well as through recent scientific research as having multiple health benefits. Claims have been made that fermented dairies such as yogurt and kefir possess anti-inflammatory, antimicrobial, antifungal, antihypertensive, and fat reducing properties (Furuno & Nakanishi, 2012). These benefits come in a tart clean dairy beverage that is thicker and creamier than unfermented dairy. This thickening is due in large part to the introduction of exopolysaccharides to the mixtures from various strains of bacteria. The term exopolysaccharide refers to a thick sugary slime emitted by certain bacterial strains. Exopolysaccharides have been heavily studied for the purpose of revealing the mechanisms for their assembly in gram-negative pathogenic bacterial strains such as Streptococcus pneumoniae, or Streptococcus pyogenes. In these cases, the substance provides a form of protection to the bacterial cell from harmful substances such as white blood cells, shifts in pH, or other noxious substances. However, exopolysaccharides are also produced by many non-pathogenic bacteria such as the lactic acid bacteria used in the production of yogurt, kefir, sour cream or non-pathogenic strains of streptococci such as Streptococcus thermophiles (Broadbent et. al, 2003). Exopolysaccharide-producing bacteria tend to produce the substance in one of two ways, capsular or non-capsular. In capsular exopolysaccaride production, the substance remains attached to the outer wall of the bacterium as an outer layer of protection. In non-capsular the exopolysaccharide substance leaves the bacterium and enters the environment surrounding the

bacterium where it produces a thicker rope-like substance. It is this occurrence that gives fermented dairy the desirable thickness and mouth-feel for which it is known (Hassan, 2008). When milk is fermenting to become yogurt a process of gelatinization takes place. As lactic acid bacteria digest the sugars in the milk, they release acids that lower the pH. This lowering of pH begins the process curdling and the milk begins to gelatinize. Casein (milk protein) micelles form creating a network that traps serums and fat in the milk (Hassan, 2008). This gelatinization occurs at a pH of 5.5. Just prior to this, as the pH is dropping from 6.6 to 5.5 conditions are ripe for increased production of exopolysaccharides from the lactic acid bacteria. These come in the non-capsular form and affect the structure of the gelatin. The ropey structures of the exopolysaccharides, when stirred, interact with each other and their presence decreases the opportunities for the proteins within the milk to interact. This creates a more open structure to the yogurt. This open structure increases the ability of water to separate out from the curd, but the exopolysaccharides have strong water retention properties, which increase the overall water holding properties of the final product. This gives the milk serum a polymer-like consistency that increases viscosity and consistency of the fermented product (Hassan, 2008). The production of exopolysaccharides from various bacteria is also being considered to improve the quality of low-fat cheeses. Traditionally, when one removes or dramatically reduces the fat content of a cheese, it loses some of its textural qualities. For example, low fat cheeses often do not melt like regular cheese. Their texture is overly hard and rubbery. This is due to the increased casein content of the cheese (Hassan, 2008). Exopolysaccharides offer a possible solution to this problem. Their quality to retain water and increase the moisture in the nonfat cheese makes exopolysaccharides an ideal addition to reduced fat cheeses. Research performed with a ropey non capsular strain of Lactococcus lactis spp. Cremoris was able to produce a 33%

reduced fat cheddar cheese that had the texture and mouth-feel of a full fat cheese. It also had a similar melting pattern to full fat cheese and was shredable (Chiang et. al, 2011). Similar results were found with reduced fat mozzarella using strains of S. theromophilus and L. bulgaricus. The increased moisture content also increased the overall yield of the cheese. (Broadbent et.al, 2003). In some cases, an exopolysaccharide can bind during fermentation with various proteins in such a way that it forms a hardened grain that builds upon itself and becomes a permanent home for diverse strains of bacteria and yeasts. This is the case with a drinkable yogurt product called kefir. True kefir contains over a dozen strains of bacteria and yeasts. Properly fermented, it has a flavor that could rival the complexity of most wines. According to Gao et. al (2012), true kefir is made from kefir grains. The grains are allowed to sit in a container filled with dairy and ferment for a period of time. The grains are then strained and the remaining liquid is imbibed as a nutrient dense probiotic. Afterwards, the grains may be reused to ferment more milk. The solid structure of the kefir grain, called kefiran, is a mixture of protein and exopolysaccharide with chain sequences of glucose and galactose residues in a hexasaccharide structure (Furuno & Nakanishi, 2012). Housed within this structure are organisms such as Leuconostic mesenteroides, Lactococcus, lactis, Lactobacillus kefiri, Lactobacillus casei, Kluyveromyces marxianus, Saccharomyces unisporus, Saccharomyces cerevisiae, and Candida humilis (Gao et. al, 2012). There appears to be a great diversity in the composition and balance of the bacteria and yeasts present from grain to grain making it difficult to translate the production of true kefir into consistent large-scale production methods (Leite et. al, 2012). Rich in lactic acid bacteria, the kefir product contains a similar mouth-feel and consistency brought about by exopolysaccharides as was present in yogurt, but the expolysaccharides in the grains make kefir a truly unique substance. Researchers have tried to no avail to reproduce these grains

synthetically. They grow and reproduce only in response to fermentation. It has been theorized that kefiran has the potential to be very useful to the food industry as a food gum, but the slow rate of growth creates a still unresolved problem for mass production (Guzel-Seydim et. al, 2011). Any existing grains of kefir are the descendants of long line of ancient grains that originated in the Caucasus Mountains where they were used to ferment goats milk inside a goatskin sack. From here they spread all over the globe. Traditions surrounding them have been found in Taiwan, Russia, Turkey, Brazil, China and more (Gao et. al, 2012). Exopolysaccharides offer a fascinating world of possibilities within food science. They can improve the texture and consistency of dairy products, produce healthier cheeses and create permanent grains for the continued fermentation and production of healthy beverages. It is possible that they have implications for food improvement outside the world of dairy. They are a natural product that we have only begun to explore in the last 20 years. Perhaps a substance that can increase the speed of kefiran generation could open the door to a natural food gum product. Research on better growth mediums is already underway (Guzel-Seydim, 2011). It appears likely, based on the current data, that the manipulation of exopolysaccharides will play a key role in the creation of food products in the near future.

References

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Chiang, M. L., Chen, H. C., Wang, S. Y., Hsieh, Y. L., & Chen, M. J. (2011). Use of taiwanese ropy fermented milk (TRFM) and lactococcus lactis subsp. cremoris isolated from TRFM in manufacturing of functional low-fat cheeses. Journal of Food Science, 76(7), M504-10. doi: 10.1111/j.1750-3841.2011.02320.x; 10.1111/j.1750-3841.2011.02320.x

Furuno, T., & Nakanishi, M. (2012). Kefiran suppresses antigen-induced mast cell activation. Biological & Pharmaceutical Bulletin, 35(2), 178-183. Gao, J., Gu, F., Abdella, N. H., Ruan, H., & He, G. (2012). Investigation on culturable microflora in tibetan kefir grains from different areas of china. Journal of Food Science, 77(8), M425-33. doi: 10.1111/j.1750-3841.2012.02805.x; 10.1111/j.17503841.2012.02805.x

Guzel-Seydim, Z., Kok-Tas, T., Ertekin-Filiz, B., & Seydim, A. C. (2011). Effect of different growth conditions on biomass increase in kefir grains. Journal of Dairy Science, 94(3), 1239-1242. doi: 10.3168/jds.2010-3349

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Leite, A. M., Mayo, B., Rachid, C. T., Peixoto, R. S., Silva, J. T., Paschoalin, V. M., & Delgado, S. (2012). Assessment of the microbial diversity of brazilian kefir grains by PCR-DGGE and pyrosequencing analysis. Food Microbiology, 31(2), 215-221. doi: 10.1016/j.fm.2012.03.011