*2.4. β-Glucuronidase Activity*

The level of β-glucuronidase (β-Glu) activity in body fluids is considered a potential biomarker in the diagnosis of certain intestinal pathological conditions [57]. Therefore, the search for potent β-Glu inhibitors in the human intestinal microflora has attracted increasing attention over the years [58], due to its role in colon carcinogenesis In particular, work is underway to discover natural dietary inhibitors of this enzyme. To date, the

main strategy for reducing or eliminating the gastrointestinal toxicity caused by bacterial β-Glu is by administration of antibiotics [59,60], but plant food and herbal medicines are a promising source of bacterial inhibitors as well [58,61,62]. To the best of our knowledge, there are no published reports on the β-Glu inhibitory activity of GF bread with insects. Our results indicate a significant ability to reduce the activity of β-Glu by CP (Table 6). The use of 6% and 10% CP in the bread recipe resulted in a decrease in the activity of β-Glu at the stage of adding the intestinal microflora to the digestive process by 63.5% and 65.9%, respectively. The β-Glu activity is also effectively inhibited in the subsequent stages of the digestive process in the large intestine. After 18 h of BCP6 and BCP10 digestion, after large intestine, the β-Glu activity was reduced by 70.6% and 78.9%, respectively. The use of food additives that inhibit β-Glu activity may also be of key importance in the treatment of certain diseases. As β-Glu plays a key role in reducing the effectiveness of anticancer drugs [63], its inhibition with food ingredients may aid the treatment of certain diseases in a non-pharmacological manner. It therefore seems that CP has the potential to be used as a new β-Glu inhibitor, and further studies of the biological activity of CP may deepen our understanding of the mechanisms underlying the beneficial effects observed in the current experiments.

**Table 6.** The results of β-glucuronidase activity after bread digestion.


Mean values with the same letters in the row (<sup>a</sup> ) were not significantly different (α = 0.05). RB—reference bread; BCP2, BCP6, BCP10—breads with starch replaced with CP at 2%, 6%, and 10%, respectively.
