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    • 3. 发明申请
    • PROCESS FOR HYDROXYLATING LONG-CHAIN ALKANES, FATTY ACIDS AND OTHER ALKYL COMPOUNDS
    • 方法羟化长链烷烃,脂肪和其他烷基化合物
    • WO1996027678A1
    • 1996-09-12
    • PCT/DE1996000410
    • 1996-03-01
    • MAX-DELBRÜCK-CENTRUM FÜR MOLEKULARE MEDIZINZIMMER, ThomasKAMINSKI, KristinaSCHUNCK, Wolf-HagenKÄRGEL, EvaSCHELLER, UlrichMAUERSBERGER, Stephan
    • MAX-DELBRÜCK-CENTRUM FÜR MOLEKULARE MEDIZIN
    • C12P07/02
    • C12N9/0042C12N9/0077C12P7/42
    • A microbial hydroxylation process is disclosed for selectively oxidising regions of long-chain alkanes, fatty acids and other alkyl compounds. The process should be easy to carry out and produce good yields of oxidation products, in particular hydroxylated fatty acids and long-chain dicarboxylic acids. The object of the invention is to modify yeast by genetic engineering so that when it is cultivated it expresses the required enzymes. The disclosed process is characterised in that the long-chain alkanes, fatty acids and other alkyl compounds are treated with monooxygenase systems that consist of cytochrom P450 and NADPOH cytochrom P450 reductase, and the hydroxylation products are then separated. The monooxygenase systems are produced in the reaction mixture by simultaneous expression of their components in yeast, preferably saccharomyces cerevisiae. The invention relates essentially to a vector for modifying saccharomyces by genetic engineering. On the basis of the structure Yep 51, the vector contains reductase cDNA between the restriction sites Sa1K and BamHI and a second expression cassette bound in the restriction site NruI. The expression cassette consists of the GAL10 promoter, the sequence that codes for cytochrom P450 and the ADH1 terminator.
    • 本发明具有用于长链烷烃,脂肪酸和其他烷基的化合物可用的区域选择性氧化提供微生物Hydroxylierungverfahren的目标。 它是在一个简单的过程管理,氧化产物的良好的产率,特别是羟基脂肪酸和长链的二羧酸,铅。 本发明具有的目的是改变遗传改造的酵母,使它们表达必需的种植酶。 本发明的方法的特征在于,所述长链烷烃,与脂肪酸和由细胞色素P450和处理过的NADPH-细胞色素P450还原酶的其他单加氧酶系统,和羟基化产物烷基化合物是分开的。 单加氧酶系统在由它的组件的在酵母中同时表达,优选在酿酒酵母中制备的反应混合物中。 本发明的关键是:酿酒的基因工程的载体,其基本骨架YEp型51中的建筑物,还原酶的限制性位点SalI和BamHI和第二表达盒连接入限制性位点的NruⅠ和由GAL10启动子,编码序列之间的cDNA 包括细胞色素P450和ADH1终止。