Yeast has been widely used for large-scale production of terpenoids. In yeast, modifications of terpenoid biosynthetic pathways have been intensively studied. tHMG1 (encoding the catalytic domain of 3-hydroxy-3-methylglutaryl coenzyme A reductase of yeast) and UPC2-1 (the G888D mutant of UPC2 encoding a transcription factor) were integrated into yeast chromosome, and ERG9 (the squalene synthase gene of yeast) was knocked down to yield the chassis strain DH02. A F96C mutation in ERG20 (farnesyl diphosphate synthase of yeast) was conducted to obtain mERG20 which can function as a geranylgeranyl diphosphate synthase (GGPS). Then, three fused genes, including BTS1 (the yeast innate GGPS)-ERG20, ERG20-mERG20 and mERG20-ERG20, were constructed, and expressed either by the pESC-based plasmids in DH02, or by being integrated into DH02 chromosome. The highest geranylgeraniol (GGOH) content was observed in the extracts of DH12 integrated with ERG20-mERG20, corresponding to 3.2 and 2.3 folds of those of the strains integrated with BTS1 and mERG20, respectively. Finally, three genes encoding nor-copalyl diphosphate synthase (nor-CPS), ent-CPS and syn-CPS were integrated into the chromosome of DH12, respectively, to construct yeasts for producing corresponding copalyl diphosphates (CPPs). Thus, a yeast-based platform was built for characterizing all types of diterpene synthases using GGPP or various CPPs as their substrates.

中文翻译：

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通过过表达ERG20及其突变体mERG20的融合酶来增强酵母中二萜的产生。

酵母已被广泛用于萜类化合物的大规模生产。在酵母中，对萜类生物合成途径的修饰已进行了深入研究。将tHMG1（编码酵母的3-羟基-3-甲基戊二酰辅酶A还原酶的催化结构域）和UPC2-1（编码转录因子的UPC2的G888D突变体）整合到酵母染色体中，并将​​ERG9（酵母的角鲨烯合酶基因）整合）被敲低产生底物菌株DH02。进行ERG20（酵母的法呢基二磷酸合酶）中的F96C突变以获得可以用作香叶基香叶基二磷酸合酶（GGPS）的mERG20。然后，构建了三个融合基因，包括BTS1（酵母先天性GGPS）-ERG20，ERG20-mERG20和mERG20-ERG20，并通过DH02中基于pESC的质粒表达或整合到DH02染色体中。在与ERG20-mERG20整合的DH12提取物中观察到最高的香叶基香叶醇（GGOH）含量，分别相当于与BTS1和mERG20整合的菌株的3.2倍和2.3倍。最终，将编码正戊基磷酸二磷酸合酶（nor-CPS），ent-CPS和syn-CPS的三个基因分别整合到DH12染色体中，以构建用于产生相应的棕榈酸二磷酸酯（CPPs）的酵母。因此，建立了基于酵母的平台，以使用GGPP或各种CPP作为底物表征所有类型的二萜合酶。分别将编码正戊二磷酸二磷酸合酶（nor-CPS），ent-CPS和syn-CPS的三个基因分别整合到DH12染色体中，以构建用于产生相应的椰油酰二磷酸（CPPs）的酵母。因此，建立了基于酵母的平台，以使用GGPP或各种CPP作为底物表征所有类型的二萜合酶。分别将编码正戊基磷酸二磷酸合酶（nor-CPS），ent-CPS和syn-CPS的三个基因整合到DH12染色体中，以构建用于产生相应的棕榈酸二磷酸酯（CPPs）的酵母。因此，建立了基于酵母的平台，以使用GGPP或各种CPP作为底物表征所有类型的二萜合酶。