Sugar nucleotide-dependent (Leloir) glycosyltransferases are powerful catalysts for glycoside synthesis. Their applicability can be limited due to elaborate production of enzyme preparations deployable in biocatalytic processes. Here, we show that efficient enzyme formulation promotes glycosyltransferases for the synthesis of the natural C-glycoside nothofagin. Adding Brij-35 detergent (1 %, w/v) during sonication of the E. coli BL21-Gold (DE3) expression strain, recovery of Oryza sativa C-glycosyltransferase was enhanced by ∼3-fold, partly due to the release of enzyme activity trapped in insoluble pellet. Freeze drying of the resulting cell-free extract (∼17 U ml⁻¹) reduced the volume ∼20-fold and gave ∼55 mg solids ml⁻¹ liquid processed, with 83 % retention of the original activity and a specific activity of 0.20 U mg⁻¹ solids. The Glycine max sucrose synthase was processed analogously, giving a solid enzyme preparation of 0.28 U mg^-1 in 63 % yield. Both enzyme formulations were stable for several weeks. The glycosyltransferase cascade reaction for 3′-β-C-glucosylation of phloretin (60 mM; as inclusion complex with hydroxypropyl-β-cyclodextrin) from UDP-glucose (generated in situ by sucrose synthase from 500 mM sucrose and 0.5 mM UDP) showed excellent performance metrics (≥ 98 % yield; 3.2 g l⁻¹ h⁻¹ space-time yield; ∼90 regeneration cycles for UDP). Collectively, our study demonstrates a facile procedure for solid glycosyltransferase formulations practically usable in glycoside synthesis.

糖核苷酸依赖性（Leloir）糖基转移酶是糖苷合成的强大催化剂。由于精心制备可在生物催化过程中使用的酶制剂，其应用可能受到限制。在这里，我们表明有效的酶制剂可促进糖基转移酶的合成，用于合成天然C-糖苷诺夫他汀。在大肠杆菌BL21-Gold（DE3）表达菌株的超声处理过程中添加Brij-35去污剂（1％，w / v），稻的C-糖基转移酶的回收率提高了约3倍，部分是由于释放了酶活性陷于不溶性沉淀物中。冷冻干燥所得的无细胞提取物（〜17 U ml ^-1），体积减少约20倍，固体含量约55 mg ml处理了^-1液体，保留了原始活性的83％，比活为0.20 U mg ^-1固体。所述大豆蔗糖合酶类似地处理，得到0.28ü毫克的固体酶制剂^-1产率为63％。两种酶制剂稳定数周。显示了来自UDP-葡萄糖（由500 mM蔗糖和0.5 mM UDP的蔗糖合酶原位产生）的促视黄素3'-β-C-葡萄糖基化（为60 mM;与羟丙基-β-环糊精的包合物）的糖基转移酶级联反应优异的性能度量（≥98％产率; 3.2 GL ^-1 ħ ^-1时空产量 UDP约为90个再生周期）。总体而言，我们的研究证明了可用于糖苷合成的固体糖基转移酶制剂的简便方法。