Recombinant Lipase from Gibberella zeae Exhibits Broad Substrate Specificity: A Comparative Study on Emulsified and Monomolecular Substrate.
Recombinant Lipase from Gibberella zeae Exhibits Broad Substrate Specificity: A Comparative Study on Emulsified and Monomolecular Substrate.
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来自玉米赤霉的重组脂肪酶表现出广泛的底物特异性：乳化底物和单分子底物的比较研究
DOI：
10.3390/ijms18071535
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发表时间：
2017-07-18
影响因子：
5.6
通讯作者：
Wang Y
中科院分区：
文献类型：
--
作者：
Wang F;Zhang H;Zhao Z;Wei R;Yang B;Wang Y
采用经典乳化体系和单分子膜技术，对源自玉米赤霉素的重组玉米赤霉素脂肪酶（rGZEL）的底物特异性进行了详细表征。在乳化反应体系下，除脂肪酶活性外，磷脂酶和糖脂酶均有水解活性。确定了不同活动的最佳展示条件。与脂肪酶活性比较，其糖脂水解活性（0.06）略高于磷脂酶活性（0.02）。rGZEL偏爱中链甘油三酯，而较长链甘油三酯活性较低。利用单分子膜技术,我们发现rGZEL的偏好顺序不同的磷脂是1,2-diacyl-sn-glycero-3-phospho-l-serine (PS) > 1, 2-dioleoyl-sn-glycero-3-phospho-rac——(1-glycerol)钠盐(PG) > 1, 2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) > l -α磷脂酰肌醇(PI) >心磷脂(CL) > 3-sn-phosphatidic酸钠盐(PA) > l -α磷脂酰乙醇胺(PE),虽然没有水解活动检测鞘磷脂(SM)。此外，rGZEL对1,2-二硬脂酰单甘油酯（MGDG）具有较高的半乳糖脂酶活性。用三对假双甘油酯对映体（ddg）对rGZEL的立体选择性和区域选择性进行了动力学研究。rGZEL对远端DDG对映体的水解活性高于邻近的酯基，但对双甘油酯类似物的sn-2位没有水解活性。此外，rGZEL更倾向于DDG对映体的R构型。分子对接结果与体外实验结果一致。
Using the classical emulsified system and the monomolecular film technique, the substrate specificity of recombinant Gibberella zeae lipase (rGZEL) that originates from Gibberella zeae was characterized in detail. Under the emulsified reaction system, both phospholipase and glycolipid hydrolytic activities were observed, except for the predominant lipase activity. The optimum conditions for different activity exhibition were also determined. Compared with its lipase activity, a little higher ratio of glycolipid hydrolytic activity (0.06) than phospholipase activity (0.02) was found. rGZEL preferred medium chain-length triglycerides, while lower activity was found for the longer-chain triglyceride. Using the monomolecular film technique, we found that the preference order of rGZEL to different phospholipids was 1,2-diacyl-sn-glycero-3-phospho-l-serine (PS) > 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (PG) > 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) > l-α-phosphatidylinositol (PI) > cardiolipin (CL) > 3-sn-phosphatidic acid sodium salt (PA) > l-α-phosphatidylethanolamine (PE), while no hydrolytic activity was detected for sphingomyelin (SM). Moreover, rGZEL showed higher galactolipase activity on 1,2-distearoyimonoglactosylglyceride (MGDG). A kinetic study on the stereo- and regioselectivity of rGZEL was also performed by using three pairs of pseudodiglyceride enantiomers (DDGs). rGZEL presented higher preference for distal DDG enantiomers than adjacent ester groups, however, no hydrolytic activity to the sn-2 position of diglyceride analogs was found. Furthermore, rGZEL preferred the R configuration of DDG enantiomers. Molecular docking results were in concordance with in vitro tests.
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影响因子：
3.4
作者：
Gutierrez-Ayesta, Cecilia;Carelli, Amalia A.;Ferreira, Maria L.
通讯作者：
Ferreira, Maria L.
影响因子：
7.2
作者：
Voigt, CA;Schäfer, W;Salomon, S
通讯作者：
Salomon, S
影响因子：
2
作者：
Rogalska, E;Nury, S;Verger, R
通讯作者：
Verger, R
DOI：
10.1111/j.1432-1033.1979.tb19736.x
发表时间：
1979-01-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
SIEBERTZ, HP;HEINZ, E;DOUCE, R
通讯作者：
DOUCE, R
影响因子：
3
作者：
Ramrakhiani, Lata;Chand, Subhash
通讯作者：
Chand, Subhash