Hasnaoui Ismail, Mechri Sondes, Dab Ahlem, Bentouhami Nour Eddine, Abouloifa Houssam, Bellaouchi Reda, Allala Fawzi, Saalaoui Ennouamane, Jaouadi Bassem, Noiriel Alexandre, Asehraou Abdeslam, Abousalham Abdelkarim
Génie Enzymatique, Membranes Biomimétiques et Assemblages Supramoléculaires (GEMBAS), Institut de Chimie et de Biochimie Moléculaires et Supramoléculaires (ICBMS), UMR 5246 CNRS, Univ Lyon, Université Lyon 1, Bât Raulin, 43 Bd du 11 Novembre 1918, F-69622 Villeurbanne cedex, France.
Laboratoire de Bioressources, Biotechnologie, Ethnopharmacologie et Santé (LBBES), Faculté des Sciences d'Oujda (FSO), Université Mohammed Premier (UMP), Bd Mohamed VI BP 717, Oujda 60000, Morocco.
Molecules. 2025 Jan 21;30(3):434. doi: 10.3390/molecules30030434.
This work describes the immobilization and the characterization of purified Thom P22 lipase (PCrL) using adsorption, encapsulation, and adsorption-encapsulation approaches. The maximum activity of the immobilized PCrL on CaCO microspheres and sodium alginate beads was shifted from 37 to 45 °C, compared with that of the free enzyme. When sodium alginate was coupled with zeolite or chitosan, the immobilization yield reached 100% and the immobilized PCrL showed improved stability over a wide temperature range, retaining all of its initial activity after a one-hour incubation at 60 °C. The immobilization of PCrL significantly improves its catalytic performance in organic solvents, its pH tolerance value, and its thermal stability. Interestingly, 95% and almost 50% of PCrL's initial activity was retained after 6 and 12 cycles, respectively. The characteristics of all PCrL forms were analyzed by X-ray diffraction and scanning electron microscopy combined with energy dispersive spectroscopy. The maximum conversion efficiency of oleic acid and methanol to methyl esters (biodiesel), by PCrL immobilized on CaCO, was 65% after a 12 h incubation at 40 °C, while free PCrL generated only 30% conversion, under the same conditions.
这项工作描述了使用吸附、包封和吸附-包封方法对纯化的汤姆P22脂肪酶（PCrL）进行固定化及表征。与游离酶相比，固定化PCrL在碳酸钙微球和海藻酸钠珠上的最大活性温度从37℃移至45℃。当海藻酸钠与沸石或壳聚糖偶联时，固定化产率达到100%，且固定化PCrL在较宽温度范围内表现出更高的稳定性，在60℃孵育1小时后仍保留其所有初始活性。PCrL的固定化显著提高了其在有机溶剂中的催化性能、pH耐受值和热稳定性。有趣的是，分别经过6次和12次循环后，PCrL仍分别保留了95%和近50%的初始活性。通过X射线衍射、扫描电子显微镜结合能谱分析了所有PCrL形式的特性。在40℃孵育12小时后，固定在碳酸钙上的PCrL将油酸和甲醇转化为甲酯（生物柴油）的最大转化效率为65%，而在相同条件下，游离PCrL的转化率仅为30%。
