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    • 10. 发明申请
    • MICROORGANISMS FOR DITERPENE PRODUCTION
    • 微生物生产的微生物
    • WO2014191581A2
    • 2014-12-04
    • PCT/EP2014061399
    • 2014-06-02
    • DSM IP ASSETS BV
    • BOER VIKTOR MARIUSSUIR ERWIN
    • A23L27/30C12P15/00
    • C12P19/56A23L2/60A23L27/36A23V2002/00C07H15/256C07K14/39C07K14/395C12N9/0036C12N9/0042C12N9/0073C12N9/0083C12N9/1051C12N9/16C12N9/2405C12N9/88C12N9/90C12N15/52C12P15/00
    • The present invention relates to a recombinant microorganism comprising one or more nucleotide sequence(s) encoding: a polypeptide having ent-copalyl pyrophosphate synthase activity;a polypeptide having ent-Kaurene synthase activity; a polypeptide having ent-Kaurene oxidase activity; and a polypeptide having kaurenoic acid 13-hydroxylase activity, whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least steviol, and wherein said recombinant microorganism has been modified in its genome such that it results in a deficiency in the production of one or more of: (i) a phosphatase capable of acting on geranylgeranylpyrophosphate (GGPP) resulting in the formation of geranylgeraniol (GOH); (ii) a phosphatase capable of acting on farnesylpyrophosphate (FPP) resulting in the formation of farnesol and nerolidol;(iii) an exo-1,3-&bgr;glucanase;(iv)a glycogen synthase (or a polypeptide that influences glycogen accumulation); (v) a transcriptional repressor of hypoxic genes (ROX1) (vi) an NADPH oxidase; or (vii) a monocarboxylate transporter (JEN1) (viii) a polypeptide having activity as encoded for by the open reading frame, YJL064w; or (ix) a polypeptide The recombinant microorganism may also be capable of expressing one or more UDP-glucosyltransferases such that the microorganism is capable of producing one or more steviol glycosides.
    • 本发明涉及包含一个或多个核苷酸序列的重组微生物,所述核苷酸序列编码:具有入 - copalyl焦磷酸合酶活性的多肽;具有ent-Kaurene合酶活性的多肽; 具有内皮氧化酶活性的多肽; 和具有海藻酸13-羟化酶活性的多肽,由此所述核苷酸序列的表达赋予所述微生物至少产生甜菊醇的能力,并且其中所述重组微生物在其基因组中已被修饰,从而得到结果 (i)能够作用于香叶基香叶基焦磷酸盐(GGPP)的磷酸酶导致形成香叶基香叶醇(GOH)的一种或多种的缺乏; (ii)能够作用于法呢碱焦磷酸(FPP)的磷酸酶,导致法呢醇和橙花叔醇的形成;(iii)外切1,3-葡聚糖酶;(iv)糖原合酶(或影响糖原积累的多肽) ); (v)缺氧基因转录抑制子(ROX1)(vi)NADPH氧化酶; 或(vii)单羧酸转运蛋白(JEN1)(viii)具有由开放阅读框编码的活性的多肽YJL064w; 或(ix)多肽重组微生物还可能能够表达一种或多种UDP-葡糖基转移酶,使得微生物能够产生一种或多种甜菊糖苷。