Release of Soybean Isoflavones by Using a β‐Glucosidase from Alicyclobacillus herbarius

β‐Glucosidases are used in the food industry to hydrolyse glycosidic bonds in complex sugars, with enzymes sourced from extremophiles better able to tolerate the process conditions. In this work, a novel β‐glycosidase from the acidophilic organism Alicyclobacillus herbarius was cloned and heterologously expressed in Escherichia coli BL21(DE3). AheGH1 was stable over a broad range of pH values (5–11) and temperatures (4–55 °C). The enzyme exhibited excellent tolerance to fructose and good tolerance to glucose, retaining 65 % activity in the presence of 10 % (w/v) glucose. It also tolerated organic solvents, some of which appeared to have a stimulating effect, in particular ethanol with a 1.7‐fold increase in activity at 10 % (v/v). The enzyme was then applied for the cleavage of isoflavone from isoflavone glucosides in an ethanolic extract of soy flour, to produce soy isoflavones, which constitute a valuable food supplement, full conversion was achieved within 15 min at 30 °C. Tough guy: Enzymatic characterisation showed that a β‐glycosidase from Alicyclobacillus herbarius was stable over a broad range of pH values and temperatures, also exhibiting excellent tolerance to sugars and organic solvents, specially to ethanol. The enzyme was applied in the hydrolysis of isoflavone glucosides in an ethanolic extract of soy flour with outstanding results.