Kuroda Hisao, Oshima Toshiyuki, Kaneda Hirotaka, Takashio Masachika
Frontier Laboratories of Value Creation, SAPPORO BREWERIES LTD., Shizuoka, Japan.
Biosci Biotechnol Biochem. 2005 Aug;69(8):1545-54. doi: 10.1271/bbb.69.1545.
Fatty acid hydroperoxide lyase (HPL), a member of cytochrome P450 (CYP74), produces aldehydes and oxo-acids involved in plant defensive reactions. In monocots, HPL that cleaves 13-hydroperoxides of fatty acids has been reported, but HPL that cleaves 9-hydroperoxides is still unknown. To find this type of HPL, in silico screening of candidate cDNA clones and subsequent functional analyses of recombinant proteins were performed. We found that AK105964 and AK107161 (Genbank accession numbers), cDNAs previously annotated as allene oxide synthase (AOS) in rice, are distinctively grouped from AOS and 13-HPL. Recombinant proteins of these cDNAs produced in Escherichia. coli cleaved both 9- and 13-hydroperoxide of linoleic and linolenic into aldehydes, while having only a trace level of AOS activity and no divinyl ether synthase activity. Hence we designated AK105964 and AK107161 OsHPL1 and OsHPL2 respectively. They are the first CYP74C family cDNAs to be found in monocots.
脂肪酸氢过氧化物裂解酶(HPL)是细胞色素P450(CYP74)家族的一员,可产生参与植物防御反应的醛类和氧代酸。在单子叶植物中,已报道了可裂解脂肪酸13-氢过氧化物的HPL,但可裂解9-氢过氧化物的HPL仍然未知。为了找到这类HPL,我们进行了候选cDNA克隆的电子筛选以及重组蛋白的后续功能分析。我们发现,AK105964和AK107161(Genbank登录号)这两个在水稻中先前被注释为丙二烯氧化物合酶(AOS)的cDNA,与AOS和13-HPL明显不同。在大肠杆菌中产生的这些cDNA的重组蛋白可将亚油酸和亚麻酸的9-和13-氢过氧化物裂解为醛类,同时仅具有微量的AOS活性且无二乙烯基醚合酶活性。因此,我们分别将AK105964和AK107161命名为OsHPL1和OsHPL2。它们是在单子叶植物中发现的首批CYP74C家族cDNA。