School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Hegeng Biotech Engineering Co., Ltd., Chuzhou 239000, China.
Food Chem. 2023 Dec 15;429:136987. doi: 10.1016/j.foodchem.2023.136987. Epub 2023 Jul 24.
A novel enzymatic process was established for galactooligosaccharides (GOS) synthesis by using plant-derived galactose as substrate, without producing any byproducts. The galactose was prepared from the acid hydrolysate of gum arabic. The yeast Kluyveromyces lactis producing β-galactosidase capable of catalyzing GOS synthesis from galactose was screened out. The synthesis conditions using the yeast cells as enzyme source were optimized by both single-factor experiment and response surface methodology, with the highest GOS yield reached 45%. The composition of reaction mixture contained only GOS and unreacted galactose, which could be easily separated by the cation exchange resin column. The structures of major GOS products were identified as Gal-β-D-(1 → 6)-Gal, Gal-β-D-(1 → 3)-Gal, and Gal-β-D-(1 → 6)-Gal-β-D-(1 → 6)-Gal by MS and NMR spectra. Moreover, the β-galactosidase-containing cells can be recycled for at least 30 batches of GOS synthesis at 35 °C, with the enzyme activity remaining above 60%.
建立了一种新型的酶法工艺,以植物来源的半乳糖为底物合成半乳糖低聚糖(GOS),不产生任何副产物。半乳糖是由阿拉伯胶的酸水解产物制备的。筛选出能够催化半乳糖合成 GOS 的产β-半乳糖苷酶的酵母 Kluyveromyces lactis。通过单因素实验和响应面法优化了使用酵母细胞作为酶源的合成条件,最高 GOS 产率达到 45%。反应混合物的组成仅含有 GOS 和未反应的半乳糖,可通过阳离子交换树脂柱轻松分离。主要 GOS 产物的结构通过 MS 和 NMR 光谱鉴定为 Gal-β-D-(1→6)-Gal、Gal-β-D-(1→3)-Gal 和 Gal-β-D-(1→6)-Gal-β-D-(1→6)-Gal。此外,含β-半乳糖苷酶的细胞在 35°C 下可回收用于至少 30 批 GOS 合成,酶活性保持在 60%以上。