Whole cell biosynthesis of luteolin glycosides by engineered Corynebacterium glutamicum harboring the amylosucrase gene

Abstract The transglycosylation activity of Deinococcus geothermalis amylosucrase (DgAS) expressed in Corynebacterium glutamicum (cDgAS) was optimized for luteolin glycoside synthesis using purified enzymes (PEs), whole cells (WCs) suspended in buffer (WCB) and WCs in ethanol (WCE). Unlike PE, both WCB and WCE had high transglycosylation yields exceeding 95% over a wide temperature range, with cells in ethanol exhibiting the greatest enzymatic activity for up to 48 h due to membrane degradation and the slow release of cDgAS, which led to lysis and death. Meanwhile, in buffer, protection of the cytoplasmic membrane and ideal pH conditions maintained activity for over 200 h. To improve their performance, WCs were immobilized on agarose and the resulting immobilized cells (ICs) had high stability in organic solvents, although WCB possessed greater thermal stability at 50 °C. The reaction parameters for PE, WCE, WCB, and IC transglycosylation were optimized using response surface methodology, and ICs required lower concentrations of substrates while synthesizing higher concentrations of reaction products in addition to producing the novel compound luteolin maltotrioside after several cycles of reuse. This study expanded on the range of possibilities for the industrial application of C. glutamicum in increasing the production and bioavailability of novel and improved products.