Full
Outline
To investigate the conditions for inducing expression levels of α-L-rhamnose glycosidase RhaB2 and its conversion of rutin, thereby enhancing the conversion rate of rutin to isoquercitrin.
Single-factor experiments examined the effects of induction temperature, duration and inducer concentration on enzyme expression levels. The effects of pH, temperature, incubation time and enzyme loading on RhaB2-mediated rutin conversion were also investigated.
Optimal expression conditions for recombinant α-L-rhamnose glycosidase RhaB2 in Escherichia coli BL21(DE3) were: Induction temperature 28 °C;induction time 20 hours; IPTG concentration 0.1 mmol/L. The optimal temperature for rutin conversion was 50 °C,with an optimal pH of 5.0. Under these conditions, a 2-hour reaction at a 2:1 enzyme-to-substrate mass ratio achieved a conversion rate of 99.96%.
In this study, a highly efficient heterologous expression system for recombinant α-L-rhamnosidase RhaB2 is successfully constructed. Under the technological conditions where the mass ratio of the enzyme to the substrate is 2:1 and the reaction time is 2 hours, the conversion rate of rutin can reach 99.96%, thus enabling the highly efficient and targeted preparation of isorhamnetin.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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