Identification of a novel phospholipase D with high transphosphatidylation activity and its application in synthesis of phosphatidylserine and DHA-phosphatidylserine
Autor: | Liu Qianqian, Lujia Zhang, Yongqian Qiu, Changhu Xue, Qingqing Han, Liu Yanjun, Xiaoqin Fan, Xiangzhao Mao |
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Rok vydání: | 2017 |
Předmět: |
0106 biological sciences
0301 basic medicine Docosahexaenoic Acids Bioengineering Phosphatidylserines 01 natural sciences Applied Microbiology and Biotechnology 03 medical and health sciences Hydrolysis chemistry.chemical_compound 010608 biotechnology Phosphatidylcholine Acinetobacter radioresistens Escherichia coli Phospholipase D biology Acinetobacter General Medicine Phosphatidic acid Phosphatidylserine biology.organism_classification Recombinant Proteins Molecular Docking Simulation 030104 developmental biology chemistry Biochemistry Docking (molecular) Thermodynamics lipids (amino acids peptides and proteins) Selectivity Biotechnology Protein Binding |
Zdroj: | Journal of biotechnology. 249 |
ISSN: | 1873-4863 |
Popis: | Phosphatidylserine (PS) and docosahexaenoic acid-phosphatidylserine (DHA-PS) have significant nutritional and biological functions, which are extensively used in functional food industries. Phospholipase D (PLD)-mediated transphosphatidylation of phosphatidylcholine (PC) or DHA-PC with l-serine, is an effective method for PS and DHA-PS preparation. However, because of the hydrolysis activity of PLD, PC and DHA-PC would be converted to the undesirable byproduct, phosphatidic acid (PA) and DHA-PA. In this study, a novel phospholipase D (PLDa2) was firstly cloned from Acinetobacter radioresistens a2 with high transphosphatidylation activity and no hydrolysis activity. In the PLD-catalyzed synthesis process (12h), both the transphosphatidylation conversion rate and selectivity of PS and DHA-PS were about 100%, which is the only PLD enzyme reported with this superiority up till now. In comparison with the majority of other known PLDs, PLDa2 exerted the highest activity at neutral pH, and it was stable from pH 4.0 to pH 9.0. In addition, PLDa2 had excellent thermal stability, with an optimum reaction temperature of 40°C and keeping more than 80% activity from 20°C to 60°C. The high catalytic selectivity mechanism of PLDa2 was explained by utilizing homology modeling, two-step docking, and binding energy and conformation analysis. PLDa2 ensured a stable supply of the biocatalyst with its most preponderant transphosphatidylation activity and PS selectivity, and had great potential in phospholipids industrial production. |
Databáze: | OpenAIRE |
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