Leibniz Universitat Hannover, Institut fur Lebensmittelchemie, Callinstratsse 5, D-30167 Hannover, Germany.
J Agric Food Chem. 2009 Nov 11;57(21):9951-5. doi: 10.1021/jf901438m.
To biotechnologically produce norisoprenoid flavor compounds, two extracellular peroxidases (MsP1 and MsP2) capable of degrading carotenoids were isolated from the culture supernatants of the basidiomycete Marasmius scorodonius (garlic mushroom). The encoding genes were cloned from genomic DNA and cDNA libraries, and databank homology searches identified MsP1 and MsP2 as members of the so-called "DyP-type" peroxidase family. Wild type enzymes and recombinant peroxidases expressed in Escherichia coli were employed for the release of norisoprenoids from various terpenoid precursor molecules. Carotenes, xanthophylls, and apocarotenals were subjected to the enzymatic degradation. Released volatile products were characterized by GC-FID and GC-MS, whereas nonvolatile breakdown products were analyzed by means of HPLC-DAD and HPLC-MS. C13 norisoprenoids together with C10 products proved to be the main volatile degradation products in each case.
为了生物技术生产法呢醇类香料化合物,从担子菌鳞伞(大蒜蘑菇)的培养液中分离出两种能够降解类胡萝卜素的细胞外过氧化物酶(MsP1 和 MsP2)。从基因组 DNA 和 cDNA 文库中克隆了编码基因,并通过数据库同源性搜索将 MsP1 和 MsP2 鉴定为所谓的“DyP 型”过氧化物酶家族的成员。野生型酶和在大肠杆菌中表达的重组过氧化物酶被用于从各种萜烯前体分子中释放法呢醇类。对类胡萝卜素、叶黄素和类胡萝卜素醛进行了酶促降解。释放的挥发性产物通过 GC-FID 和 GC-MS 进行了表征,而非挥发性的降解产物则通过 HPLC-DAD 和 HPLC-MS 进行了分析。在每种情况下,C13 法呢醇类化合物以及 C10 产物被证明是主要的挥发性降解产物。