Purification, identification, characterization and catalytic mechanism of two lipases from rice bran (Oryza sativa)
Purification, identification, characterization and catalytic mechanism of two lipases from rice bran (Oryza sativa)
复制标题
两种米糠脂肪酶的纯化、鉴定、表征及催化机制
DOI：
10.1016/j.lwt.2020.110693
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发表时间：
2021-04
期刊：
影响因子：
--
通讯作者：
Deng Ze-yuan
中科院分区：
文献类型：
--
作者：
Yu Cheng-wei;Zheng Liu-feng;Cheng Ming-yan;Yu Xin-ying;Wang Shu-yi;Fan Ya-wei;Deng Ze-yuan
由于米糠脂肪酶的快速酸败，严重阻碍了米糠的高效利用，因此迫切需要对米糠脂肪酶的特性和催化机理进行研究。从米糠中提取粗蛋白，经纯化得到两种脂肪酶，纯化倍数为16.31倍，纯化得率为10.64%。这两种同源脂肪酶的分子量分别为34929.00 Da和34072.00 Da。米糠脂肪酶在40.00 °C、pH7.50时具有最高的催化活性，在50.00 °C以下和pH7.00 ~ 7.50范围内具有较好的稳定性。大多数金属离子（除Na+和Co+外）和有机溶剂对纯化的脂肪酶活性有显著的抑制作用（P< 0.05）。表面活性剂Tween 80、Triton 100和十二烷基磺酸钠（SDS）使脂肪酶活性显著降低（P< 0.05），分别为70.88%、80.08%和73.74%，十六烷基三甲基溴化铵（CTAB）使酶活性显著提高（P< 0.05），为109.00%。利用针对特定氨基酸的抑制剂（苯甲磺酰氟（PMSF）、焦碳酸二乙酯（DEPC）和胃蛋白酶抑制素（Pepstatin）），推测Ser-His-Asp残基可能是纯化脂肪酶的催化中心，并通过分子对接进行验证。因此，阻断或破坏Ser-His-Asp催化三联体可能是抑制米糠酸败的新选择。
The efficient utilization of rice bran is severely impeded because of the rapid rancidity caused by rice bran lipase, so the investigation of rice bran lipase characteristic and catalytic mechanism was urgently required. The extracted crude proteins from rice bran were purified to obtain two lipases with 16.31 times of purification fold and 10.64% of purification yield. The molecular masses of these two homologous lipases were 34929.00 Da and 34072.00 Da, respectively. Rice bran lipase showed highest catalytic activity at 40.00 °C, pH 7.50 and held good stability below 50.00 °C and at pH 7.00–7.50. Most metal ions (except Na+and Co+) and organic solvents had significant inhibition effect (P< 0.05) on the purified lipase activity. Surfactants like Tween 80, Triton 100 and sodium dodecyl sulfonate (SDS) decreased the lipase activities significantly (P< 0.05) to 70.88%, 80.08% and 73.74% respectively and cetyltrimethyl ammonium bromide (CTAB) increased the catalytic activity significantly (P< 0.05) to 109.00%. The Ser-His-Asp residues were speculated to be the catalytic center of purified lipases by using the inhibitors (Phenylmethanesulfonyl fluoride (PMSF), diethylpyrocarbonate (DEPC) and Pepstatin) which aimed at specific amino acids and then verified by molecular docking. Therefore, blocking or destroying the Ser-His-Asp catalytic triad might be new choices to restrain the rancidity of rice bran.
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影响因子：
1.4
作者：
Dhingra, Devinder;Chopra, Sangeeta;Rai, D. R.
通讯作者：
Rai, D. R.
影响因子：
3.9
作者：
KIM, CJ;BYUN, SM;KWON, TW
通讯作者：
KWON, TW
DOI：
10.1016/j.lwt.2019.108990
发表时间：
2020-03
期刊：
LWT
影响因子：
--
作者：
Liufeng Zheng;Xinying Yu;Changhao Wei;Leyun Qiu;Chengwei Yu;Qian Xing;Ya-wei Fan;Z. Deng
通讯作者：
Liufeng Zheng;Xinying Yu;Changhao Wei;Leyun Qiu;Chengwei Yu;Qian Xing;Ya-wei Fan;Z. Deng
影响因子：
3.8
作者：
C. S. María;E. Revilla;B. Rodríguez-Morgado;A. Castaño;P. Carbonero;B. Gordillo;R. Cert;J. Parrado
通讯作者：
C. S. María;E. Revilla;B. Rodríguez-Morgado;A. Castaño;P. Carbonero;B. Gordillo;R. Cert;J. Parrado
DOI：
10.1080/00021369.1976.10862049
发表时间：
1976-02
期刊：
Agricultural and biological chemistry
影响因子：
--
作者：
Y. Aizono;M. Funatsu;Y. Fujiki;M. Watanabe
通讯作者：
Y. Aizono;M. Funatsu;Y. Fujiki;M. Watanabe