{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T01:45:22Z","timestamp":1791423922222,"version":"4.3.3"},"reference-count":25,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2012,9,28]],"date-time":"2012-09-28T00:00:00Z","timestamp":1348790400000},"content-version":"vor","delay-in-days":271,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/3.0\/"},{"start":{"date-parts":[[2015,9,1]],"date-time":"2015-09-01T00:00:00Z","timestamp":1441065600000},"content-version":"tdm","delay-in-days":1339,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1"},{"start":{"date-parts":[[2015,9,1]],"date-time":"2015-09-01T00:00:00Z","timestamp":1441065600000},"content-version":"tdm","delay-in-days":1339,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["elsevier.com","sciencedirect.com"],"crossmark-restriction":true},"short-container-title":["FEBS Open Bio"],"published-print":{"date-parts":[[2012,1]]},"abstract":"<jats:p>A novel metal ion\u2010independent phospholipase A<jats:sub>1<\/jats:sub> of <jats:italic>Streptomyces albidoflavus<\/jats:italic> isolated from Japanese soil has been purified and characterized. The enzyme consists of a 33\u2010residue N\u2010terminal signal secretion sequence and a 269\u2010residue mature protein with a deduced molecular weight of 27,199. Efficient and extracellular production of the recombinant enzyme was successfully achieved using <jats:italic>Streptomyces lividans<\/jats:italic> cells and an expression vector. A large amount (25 mg protein, 14.7 kU) of recombinant enzyme with high specific activity (588 U\/mg protein) was purified by simple purification steps. The maximum activity was found at pH 7.2 and 50 \u00b0C. At pH 7.2, the enzyme preferably hydrolyzed phosphatidic acid and phosphatidylserine; however, the substrate specificity was dependent on the reaction pH. The enzyme hydrolyzed lysophosphatidylcholine and not triglyceride and the <jats:italic>p<\/jats:italic>\u2010nitrophenyl ester of fatty acids. At the reaction equilibrium, the molar ratio of released free fatty acids (<jats:italic>sn<\/jats:italic>\u20101:<jats:italic>sn<\/jats:italic>\u20102) was 63:37. The hydrolysis of phosphatidic acid at 50 \u00b0C and pH 7.2 gave apparent <jats:italic>V<\/jats:italic>\n<jats:sub>max<\/jats:sub> and <jats:italic>k<\/jats:italic>\n<jats:sub>cat<\/jats:sub> values of 1389 \u03bcmol min<jats:sup>\u22121<\/jats:sup> mg protein<jats:sup>\u22121<\/jats:sup> and 630 s<jats:sup>\u22121<\/jats:sup>, respectively. The apparent <jats:italic>K<\/jats:italic>\n<jats:sub>m<\/jats:sub> and <jats:italic>k<\/jats:italic>\n<jats:sub>cat<\/jats:sub>\/<jats:italic>K<\/jats:italic>\n<jats:sub>m<\/jats:sub> values were 2.38 mM and 265 mM<jats:sup>\u22121<\/jats:sup> s<jats:sup>\u22121<\/jats:sup>, respectively. Mutagenesis analysis showed that Ser11 is essential for the catalytic function of the enzyme and the active site may include residues Ser216 and His218.<\/jats:p>","DOI":"10.1016\/j.fob.2012.09.006","type":"journal-article","created":{"date-parts":[[2012,9,28]],"date-time":"2012-09-28T22:02:06Z","timestamp":1348869726000},"page":"318-327","update-policy":"https:\/\/doi.org\/10.1016\/elsevier_cm_policy","source":"Crossref","is-referenced-by-count":31,"title":["Purification, characterization, molecular cloning and extracellular production of a phospholipase A<sub>1<\/sub> from <i>Streptomyces albidoflavus<\/i> NA297"],"prefix":"10.1002","volume":"2","author":[{"given":"Daisuke","family":"Sugimori","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kota","family":"Kano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yusaku","family":"Matsumoto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2012,9,28]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1271\/bbb.63.820"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF01570000"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi00800a015"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1093\/oxfordjournals.jbchem.a132607"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M200264200"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.187.3.1192-1195.2005"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1388-1981(00)00107-4"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.toxicon.2007.06.027"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.enzmictec.2007.09.004"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.2323\/jgam.16.1_81"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(20)79778-4"},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.1271\/bbb1961.43.517"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.201000398"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(17)36208-7"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.1021\/cb5000014"},{"key":"e_1_2_7_17_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.66.3.890-894.2000"},{"key":"e_1_2_7_18_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11746-998-0309-z"},{"key":"e_1_2_7_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/0385-6380(87)90108-7"},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00241689"},{"key":"e_1_2_7_21_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01060-08"},{"key":"e_1_2_7_22_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462-5822.2007.00901.x"},{"key":"e_1_2_7_23_1","doi-asserted-by":"publisher","DOI":"10.1021\/ac950914h"},{"key":"e_1_2_7_24_1","unstructured":"T Kieser MJ Bibb MJ Buttner KF Chater DA Hopwood 2000 The John Innes Foundation; 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