B-590.19.112 [2020-10] Sucrose – Enzymatic Method

The extract content of normal beer wort is typically composed of around 90 % carbohydrates, which occupies the position of highest importance for beer production.

The starch reserves originally found in the barley kernel are converted to some extent into soluble carbohydrates by degradation during malting; however, the majority of soluble carbohydrates are released during mashing. The products of starch degradation can be classified into three groups:

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Application/Purpose

Determination of sucrose by enzymatic means

Scope of Application

Suitable for wort, beer, malt beverages, nutrient beer, mixed beer beverages, NAB, juices and beverages

Principle

Sucrose is important as a fermentable sugar for the technology of wort and beer production. Sucrose also plays a role in the evaluation and assessment of malt beverages and nutritional beers.

D-glucose content is determined before and after enzymatic hydrolysis of sucrose.

Sucrose is hydrolyzed by the enzyme β-fructosidase (invertase) at pH 4.6 to glucose and fructose:

\(\text{Sucrose + } H_2O \space {\xrightarrow{β-fructosidase}} \space \text{D-glucose + D-fructose}\)

Glucose is phosphorylated by the enzyme hexokinase (HK) and adenosine 5'-triphosphate (ATP) to glucose 6-phosphate (G-6-P):

\(\text{Glucose}+\text{ATP} \space \xrightarrow{HK} \space \text{G-6-P + ADP}\)

In the presence of the enzyme glucose-6-phosphate dehydrogenase (G6P-DH), G-6-P is oxidized from nicotinamide adenine dinucleotide phosphate (NADP) to gluconate-6-phosphate. Reduced nicotinamide adenine dinucleotide phosphate (NADPH) is formed:

\(\text{G-6-P + NADP} \hspace{0.8em} \xrightarrow{G6P-DH} \hspace{0.8em} \text{gluconate-6-phosphate + NADP + H}^+\)

The amount of NADPH formed during the reaction is equivalent to the amount of glucose. NADPH is a measurand and is determined on the basis of its absorbance at 334, 340 or 365 nm.

The sucrose content is calculated from the difference between the glucose concentration before and after enzymatic inversion.

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