Efficient acquisition of high-purity cyanidin-3-O-glucoside from mulberry fruits: An integrated process of ATPS whole-cell transformation and semi-preparative HPLC purification

• Published in: Food chemistry Vol. 404; p. 134651
• Main Authors: Li, Yi-Tong, Huang, Ting, Wang, Jin-Zheng, Yan, Cheng-Hai, Gong, Lu-Chan, Wu, Fu-An, Wang, Jun
• Format: Journal Article
• Language: English
• Published: Elsevier Ltd 15.03.2023
• Subjects: Aqueous two-phase system; Cyanidin-3-O-glucoside; Cyanidin-3-O-rutinoside; Mulberry red pigment; Semi-preparative HPLC; Whole cell; Mulberry red pigment; Semi-preparative HPLC; Whole cell; Cyanidin-3-O-rutinoside; Aqueous two-phase system; Cyanidin-3-O-glucoside
• ISSN: 0308-8146; 1873-7072
• DOI: 10.1016/j.foodchem.2022.134651

•Mulberry anthocyanins were prepared by ATPS biotransformation and HPLC purification.•An ATPS was designed to enhance the conversion of mulberry red pigment from C3R to C3G.•Whole-cell catalysts containing α-rhamnosidase at the interface of ATPS for easy recovery.•Peak partition collection and post-processing of semi-preparative HPLC was established.•The final purities of C3G and C3R prepared by semi-preparative HPLC reached 99 %. As a nutritious fruit, mulberry is an ideal source of high-quality cyanidin-3-O-glucoside (C3G) with various biological activities. However, the difficult separation process of high-purity C3G leads to its high price. To rapidly prepare high-purity C3G, cyanidin-3-O-rutinoside is converted to C3G by direct hydrolysis of rhamnose bond using a whole-cell catalyst containing α-rhamnosidase. Combined with an aqueous two-phase system, a coupling reaction separation system was established. Two monomers were successfully separated by semi-preparative high performance liquid chromatography (semi-preparative HPLC). The conversion of C3G catalyzed by whole-cells in the PEG/Na2SO4 system increased from 47.11 % to 66.56 %, compared with the EtOH/(NH4)2SO4 system, and the whole-cell activity remained above 50 % after five rounds of reuse. Meanwhile, the purity of C3G was increased to 99 % via the semi-preparative HPLC purification and identified by MS. Thus, an integrated process of whole-cell-catalyzed conversion and product peak cutting partition collection provides a novel strategy for efficient biomanufacturing of high-purity C3G.