来源于 Ramopichia ramosa 的嗜热真菌 GH36 α-半乳糖苷酶及其对槐豆胶的协同水解作用。
A thermophilic fungal GH36 α-galactosidase from Lichtheimia ramosa and its synergistic hydrolysis of locust bean gum.
机构信息
School of Biology and Biological Engineering, South China University of Technology, Guangzhou, Guangdong, 510006, China; AsiaPac (Dongguan) Bio-Technology Co., Ltd., No. 3, North Industrial Road, Songshan Lake National Hi-tech Industrial Development Zone, Dongguan City, Guangdong, 523808, China.
School of Biology and Biological Engineering, South China University of Technology, Guangzhou, Guangdong, 510006, China; Guangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, Guangzhou, Guangdong, 510006, China.
出版信息
Carbohydr Res. 2020 May;491:107911. doi: 10.1016/j.carres.2020.107911. Epub 2020 Jan 18.
A novel GH36 α-galactosidase gene (LrAgal36A) from Lichtheimia ramosa was synthesized and highly expressed in Pichia pastoris. The enzyme titer and protein yield for high-density fermentation in a 5 L fermentor were up to 953.6 U mL and 4.36 g L. Purified recombinant LrAgal36A showed the maximum activity at pH 6.0 and 65 °C and was thermostable with a half-life of 70 min at 60 °C. LrAgal36A displayed the highest specific activity (353.17 ± 4.19 U mg) toward p-nitrophenyl-α-d-galactopyranoside (pNPGal) followed by galacto-oligosaccharides and could act slightly on galactomannans. The K and catalytic efficiency (k/K) of LrAgal36A for pNPGal were 0.33 mM and 1569.50 mM s, respectively. LrAgal36A and GH5 β-mannanase from L. ramosa showed a significant synergistic effect on the degradation of locust bean gum (LBG), resulting in release more reducing sugars (1.56 folds) and galactose (7.6 folds) by simultaneous or sequential reactions. Due to its hydrolysis properties, LrAgal36A might have potential applications in the area of pulp biobleaching, feed and food processing.
从裂殖壶菌（Lichtheimia ramosa）中合成了一种新型 GH36 α-半乳糖苷酶基因（LrAgal36A），并在毕赤酵母中进行了高效表达。在 5 L 发酵罐中进行高密度发酵时，酶的效价和蛋白产量最高可达 953.6 U/mL 和 4.36 g/L。纯化的重组 LrAgal36A 在 pH 6.0 和 65°C 时表现出最大活性，在 60°C 下半衰期为 70 分钟，具有热稳定性。LrAgal36A 对 p-硝基苯-α-d-半乳糖吡喃糖苷（pNPGal）的比活力最高（353.17±4.19 U/mg），其次是半乳糖低聚糖，对半乳甘露聚糖略有作用。LrAgal36A 对 pNPGal 的 K 和催化效率（k/K）分别为 0.33 mM 和 1569.50 mM·s。裂殖壶菌的 LrAgal36A 和 GH5 β-甘露聚糖酶对罗望子豆胶（LBG）的降解表现出显著的协同作用，导致还原糖（提高 1.56 倍）和半乳糖（提高 7.6 倍）的释放量增加，通过同时或顺序反应。由于其水解特性，LrAgal36A 在纸浆生物漂白、饲料和食品加工等领域可能具有潜在的应用价值。
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