The efficient and economical conversion of agricultural waste into glycosides and rare sugars is challenging. Herein, an in vitro synthetic bienzyme system consisting of sucrose phosphorylase and d-allulose 3-epimerase was constructed to produce 2-O-α-glucosyl glycerol and d-allulose from cane molasses. Lactic acid in the cane molasses significantly induced sucrose phosphorylase to hydrolyze sucrose instead of glycosylation. Notably, lactic acid significantly inhibited the catalytic performance of d-allulose 3-epimerase only in the presence of Na⁺ and K⁺, with an inhibition rate of 75%. After removing lactic acid and metal ions, 116 g/L 2-O-α-glucosyl glycerol and 51 g/L d-allulose were synthesized from 500 mM sucrose in the treated cane molasses with a sucrose consumption rate of 97 %. Our findings offer an economically efficient and environmentally friendly pathway for the industrial production of glycosides and rare sugars from food industry waste.

将农业废物高效且经济地转化为糖苷和稀有糖具有挑战性。在此，构建了由蔗糖磷酸化酶和d-阿洛酮糖3-差向异构酶组成的体外合成双酶系统，以从甘蔗糖蜜生产2-O-α-葡萄糖基甘油和d-阿洛酮糖。甘蔗糖蜜中的乳酸显着诱导蔗糖磷酸化酶水解蔗糖而不是糖基化。^{值得注意的是，只有在Na +}和K ⁺存在的情况下，乳酸才显着抑制d-阿洛酮糖3-差向异构酶的催化性能，抑制率为75%。处理后的甘蔗糖蜜去除乳酸和金属离子后，以500 mM蔗糖为原料合成116 g/L 2-O-α-葡萄糖基甘油和51 g/L d-阿洛酮糖，蔗糖消耗率为97%。我们的研究结果为从食品工业废物中工业生产糖苷和稀有糖提供了一条经济有效且环境友好的途径。