The enzymatic pathway of xanthan depolymerization has been predicted previously; however, the β-glucosidase and unsaturated glucuronyl hydrolase in this system have not been cloned and characterized. This lack of knowledge hinders rational modification of xanthan and exploration of new applications. In this work, we report on the properties of Mibgl3, a xanthan-degrading enzyme isolated from Microbacterium sp. XT11. Mibgl3 exhibits typical structural features of the GH3 family but shares low sequence identity with reported GH3 enzymes. The activity of Mibgl3 can be inhibited by Cu2+, Fe2+, Zn2+, and glucose. Unlike most β-glucosidases, Mibgl3 can tolerate a wide pH range and is activated by high concentrations of NaCl. This improves the commercial value of Mibgl3. In particular, Mibgl3 exhibits higher substrate specificity toward oligoxanthan than other β-glucosidases. Ion chromatography, ultrahigh-performance liquid chromatography-mass spectrometry (UPLC-MS), and GC-MS results showed that Mibgl3 could effectively hydrolyze oligoxanthan to release glucose and glucuronate. Therefore, Mibgl3 might play an important role in xanthan depolymerization by functioning as hydrolase of both the xanthan backbone and sidechains. This knowledge of the enzymatic properties and hydrolysis mechanism of a β-glucosidase will be beneficial for future applications.
Keywords: bifunctional enzyme; unsaturated glucuronyl hydrolase; xanthan depolymerization; β-glucosidase.