New phospholipase A1-producing bacteria from a marine fish.
Phospholipase A1 is a hydrolytic enzyme that catalyzes the removal of the acyl group from position 1 of glycerophospholipids to form 2-acyl lysophospholipids. Lysophospholipids are used in foods, cosmetics, and pharmaceuticals as surfactants. Novel forms of phospholipase A1 that function at low temperatures are desirable for use in lipophilic systems in food processing. However, there is currently little variety in the available sources of phospholipase A1. Given this situation, we screened the intestinal contents of marine animals for phospholipase A1-producing bacteria. Colonies that formed a halo on K28CP screening medium and that grew in K28 medium were cultured in liquid K28 medium, and the supernatant was retrieved for analysis. Phosphatidylcholine was added to the culture supernatant, and the product of the reaction was analyzed by using TLC. For culture supernatants that were able to generate lysophosphatidylcholine, synthetic phosphatidylcholines were added, and the site of the reaction was determined by analyzing the fatty acid compositions of the lysophosphatidylcholines generated by GLC. A bacterial isolate from a flatfish, which we named HFKI0020, was found to have phospholipase A1 activity at low temperatures. We determined that the isolate HFKI0020 is closely related to Pseudomonas by using 16S rDNA sequence analysis and by characterizing the isolate with respect to its physiologic and biochemical properties. From the intestinal contents of a marine fish, we successfully isolated a bacterium that secretes phospholipase A1 that is active at low temperatures.
Nishihara
Masaaki
M
Laboratory of Nutraceuticals and Functional Foods Science, Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Tokyo, Japan.
Kamata
Masazumi
M
Koyama
Tomoyuki
T
Yazawa
Kazunaga
K
D000818:Animals; D005227:Fatty Acids / Q000737:chemistry / Q000378:metabolism; D005412:Flatfishes / Q000382:microbiology*; D005766:Gastrointestinal Contents / Q000382:microbiology; D008386:Marine Biology; D008969:Molecular Sequence Data; D054466:Phospholipases A1 / Q000378:metabolism*; D010802:Phylogeny; D011549:Pseudomonas / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013696:Temperature
D005227:Fatty Acids; D012336:RNA, Ribosomal, 16S; D054466:Phospholipases A1