Xuefeng Tu, Sifang Zhou, Can Yang, Fei Chang, Jie Liu, Hua Chen
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引用次数: 0
摘要
d -泛酸盐是人体必需的微量营养素之一。立体选择性d -内酯水解酶(D-lac)可将l -pantolactone水解为D-pantoic acid, D-pantoic acid是D-pantothenate的前体。本研究将念珠镰刀菌D-lac基因在大肠杆菌中异质表达。随后,采用含有重组细胞的海藻酸钠和CaCl2作为打印材料,进行同轴3D打印。采用同轴喷嘴打印中空细丝用于重组细胞的固定化。优化打印条件后,与自由细胞相比,打印的固定细胞具有更高的稳定性和更广泛的反应条件(如pH和温度)。经过20个反应循环后,酶活性维持在原酶活性的80%左右。通过扫描电子显微镜(SEM)对打印的空心细丝进行了验证。微通道结构和丝的大比表面积有利于底物交换,提高催化效率。结果表明,3D打印的中空纤维可以作为一种潜在的酶固定化材料。
Immobilizing D-lactonohydrolase with 3D-printed hollow filaments to improve the enzyme stability and reusability.
D-pantothenate is one of the essential micronutrients required by the organism. Stereoselective D-lactonohydrolase (D-lac) can hydrolyze DL-pantolactone into D-pantoic acid, the precursor of D-pantothenate. In this study, the D-lac gene from Fusarium moniliforme was heterogeneously expressed in Escherichia coli. Subsequently, coaxial 3D printing was employed, using sodium alginate containing the recombinant cells and CaCl2 as printing materials. The coaxial nozzle was used to print hollow filaments for the immobilization of recombinant cells. After optimizing the printing conditions, the printed immobilization cells exhibited higher stability and a wider range of reaction conditions (such as pH and temperature) compared to the free cells. After 20 reaction cycles, the enzyme activity maintained approximately 80% of original. The printed hollow filaments were confirmed by scanning electron microscope (SEM). The microchannel structures and large specific surface of filaments may facilitate substrate exchange and enhance catalytic efficiency. The results indicated that the 3D printed hollow filaments can be used as a potential material for enzyme immobilization.
期刊介绍:
Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them.
All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included.
Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields.
The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories.
Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.