Shin Kyung-Chul, Seo Min-Ju, Oh Deok-Kun, Choi Mi-Na, Kim Dae-Wook, Kim Yeong-Su, Park Chang-Su
Department of Bioscience and Biotechnology, Konkuk University, 1 Hwayang-dong Gwangjin-gu, Seoul, 143-701, Republic of Korea.
Plant Resource Industry Division, Baekdudaegan National Arboretum, Bonghwagun, Republic of Korea.
Biotechnol Lett. 2019 Mar;41(3):419-426. doi: 10.1007/s10529-019-02648-8. Epub 2019 Jan 21.
This study was conducted to characterize recombinant α-L-rhamnosidase from Chloroflexus aurantiacus and apply the enzyme in the production of isoquercitrin from rutin.
The α-L-rhamnosidase from C. aurantiacus was cloned and expressed in Escherichia coli BL21 and purified as a soluble enzyme. α-L-rhamnosidase purified from C. aurantiacus has a molecular mass of approximately 105 kDa and is predicted to exist as a homodimer with a native enzyme of 200 kDa. The purified enzyme exhibited the highest specific activity for rutin among the reported isoquercitrin producing α-L-rhamnosidases and was applied in the production of isoquercitrin from rutin. Under the optimised conditions of pH 6.0, 50 °C, 0.6 U mL α-L-rhamnosidase, and 30 mM rutin, α-L-rhamnosidase from C. aurantiacus produced 30 mM isoquercitrin after 2 h with a 100% conversion yield and productivity of 15 mM h.
We achieved a high productivity of isoquercitrin from rutin. Moreover, these results suggest that α-L-rhamnosidase from C. aurantiacus is an effective isoquercitrin producer.
本研究旨在对来自橙黄嗜热栖热菌的重组α-L-鼠李糖苷酶进行表征，并将该酶应用于从芦丁生产异槲皮苷。
来自橙黄嗜热栖热菌的α-L-鼠李糖苷酶在大肠杆菌BL21中克隆表达，并作为可溶性酶进行纯化。从橙黄嗜热栖热菌纯化的α-L-鼠李糖苷酶分子量约为105 kDa，预计以天然酶200 kDa的同型二聚体形式存在。在已报道的用于生产异槲皮苷的α-L-鼠李糖苷酶中，纯化后的酶对芦丁表现出最高的比活性，并应用于从芦丁生产异槲皮苷。在pH 6.0、50°C、0.6 U/mL α-L-鼠李糖苷酶和30 mM芦丁的优化条件下，来自橙黄嗜热栖热菌的α-L-鼠李糖苷酶在2小时后产生了30 mM异槲皮苷，转化率为100%，生产力为15 mM/h。
我们从芦丁中实现了异槲皮苷的高生产力。此外，这些结果表明来自橙黄嗜热栖热菌的α-L-鼠李糖苷酶是一种有效的异槲皮苷生产酶。
