Exopolysaccharide

I. W. Sutherland, Biotechnology of Microbial Exopolysaccharides, Cambridge University Press, Cambridge, U.K., 1990. 

Plant stmctural material is the polysaccharide cellulose, which is a linear P (1 — 4) linked polymer. Some stmctural polysaccharides iacorporate nitrogen iato thek molecular stmcture an example is chitin, the material which comprises the hard exoskeletons of kisects and cmstaceans. Chitki is a cellulose derivative whereki the OH at C-2 is replaced by an acetylated amino group (—NHCOCH ). Microbial polysaccharides, of which the capsular or extracellular (exopolysaccharides) are probably the most important class, show more diversity both ki monomer units and the nature of thek linkages. 

Metabolites whose biosynthesis is energy requiring, for example exopolysaccharides using certain substrates. Here, part of the substrate has to be oxidised to provide ATP for biosynthesis and thus the P/O quotient of the producing organism influences the theoretical yield. 

In this section we will consider the energetics of exopolysaccharide production in some detail. We will see how chemostat (substrate limited) derived yield coefficients and slfbstrate elemental balances can be used to determine how the nature of the substrate influences... 

The energetic requirements of exopolysaccharide production from various carbon sources can be calculated if the P/O quotient during growth on the carbon substrate is known. Table 3.1 shows molar growth yields measured during carbon limited growth in chemostat culture. 

We can see from Table 3.1 that the P/ O quotient is virtually independent of the carbon constant P/O source. We can therefore assume a constant P/O quotient when calculating the, , olient energetic consequences of exopolysaccharide production from different carbon sources. 

Table 3.1 Parameters of growth and exopolysaccharide production for Agrobacterium radiobacter grown in chemostat culture on various carbon sources. Data obtained from Linton J. D. et al (1987) Journal of General Microbiology 133, 2979-2987.

Examine Table 3.1. What is the relationship between specific rate of exopolysaccharide production and growth efficiency ... 

In order to quantify the scope for improvement of exopolysaccharide production, it is first necessary to correct the observed yields of exopolysaccharide for the amount of carbon substrate and oxygen required for cell production. The corrected yields are then compared with the theoretical calculated from the P/O quotient for the producing micro-organism. Such a comparison is made in Table 3.3. 

Table 3.3 Experimental stoichiometries for cell growth and exopolysaccharide production from various carbon sources by Agrvbacterhim radlobacter under nitrogen-limiting conditions.

The requirement for oxygen and carbon source for cell biosynthesis are calculated using nitrogen-limited mass balance equations for growth during exopolysaccharide production 01 res (nitrogen-limited cultures). These balances are derived from experimentally determined values of ... 

Experimentally determined yields of exopolysaccharide have been found to be 70% of the theoretical. This suggests that exopolysaccharide production is an efficient process with little scope for major yield improvements. 

To which class(es) of metabolite, based on the relationship between energy and metabolite synthesis, would you expect exopolysaccharides to belong Explain your reasoning. 

Commercial applications for polysaccharides include their use as food additives, medicines and industrial products. Although plant polysaccharides (such as starch, agar and alginate) have been exploited commercially for many years, microbial exopolysaccharides have only become widely used over the past few decades. The diversity of polysaccharide structure is far greater in micro-organisms compared to plants and around 20 microbial polysaccharides with market potential have been described. However, microorganisms are still considered to be a rich and as yet underexploited source of exopolysaccharides. 

Although exopolysaccharides do not normally have a structural role, they do form structures that can be detected by either light or electron microscopy. Exopolysaccharides may form part of a capsule closely attached to the microbial cell surface, or appear as loose slime secreted by the cell but not directly attached to it mucoid Exopolysaccharide producing cells usually form mucoid colonies on solid media and colonies liquid cultures of these cells may become very viscous. However, growth conditions can... 

Figure 7.1 Structures of some of the components of microbial exopolysaccharides.

The presence of uronic acids in microbial exopolysaccharides results in their polyanionic nature. 

In addition to one or more sugars, exopolysaccharides from prokaryotes commonly contain pyruvate ketals and various ester-linked organic substituents. These are only rarely found in eukaryotic exopolysaccharides. 

Pyruvate ketals add to the anionic nature of the exopolysaccharide and are usually present in stoichiometric ratios with the carbohydrate component Pyruvate is normally attached to the neutral hexoses but may also be attached to uronic adds. In the absence of uronic acids, pyruvate alone contributes to the anionic nature of the exopolysaccharide. 

Acetate is the commonest ester-linked component of exopolysaccharides and does not contribute to their anionic nature. Less common ester-linked components, which may be found along with acetate in some exopolysaccharides, include ... 

The presence of organic acid substituents in exopolysaccharides increases the lipophilidty of the molecule. In addition, for some exopolysaccharides with relatively high organic acid contents, their interaction with cations and with other polysaccharides may be influenced. Several amino adds have also been found in bacterial exopolysaccharides, including serine and L-glutamic add (Figure 7.1). 

Some microbial exopolysaccharides contain the inorganic substituents phosphate and sulphate. Phosphate has been found in exopolysaccharide from bacteria of medical importance, including Escherichia coli. Sulphate is far less common than phosphate and has only been found in spedes of cyanobaderia. In addition to these inorganic components, which form part of the structure of some exopolysaccharides, all polyanionic polymers will bind a mixture of cations. Exopolysaccharides are, therefore, purified in the salt form. The strength of binding of the various cations depend on the exopolysaccharide some bind the divalent cations calrium, barium and strontium very strongly, whereas others prefer certain monovalent cations, eg Na ... 

Pyruvate ketals contribute to the cationic nature of exopolysaccharides. 

The presence of acetate in exopolysaccharides increases their lipophilidty. 

An exopolysaccharide containing a high content of D-glucuronic add will tend to bind cations. 

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