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首页> 外文期刊>The FEBS journal >Increased NADPH concentration obtained by metabolic engineering of the pentose phosphate pathway in Aspergillus niger.
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Increased NADPH concentration obtained by metabolic engineering of the pentose phosphate pathway in Aspergillus niger.

机译：通过黑曲霉中戊糖磷酸途径的代谢工程获得的增加的NADPH浓度。

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Many biosynthetic reactions and bioconversions are limited by low availability of NADPH. With the purpose of increasing the cellular NADPH concn. and/or flux through the pentose phosphate pathway in Aspergillus niger, genes encoding glucose 6-phosphate dehydrogenase (gsdA), 6-phosphogluconate dehydrogenase (gndA) and transketolase (tktA) were cloned and overexpressed in separate strains. Intracellular NADPH concn. was increased 2-9-fold as a result of a 13-fold overproduction of 6-phosphogluconate dehydrogenase. Although overproduction of glucose 6-phosphate dehydrogenase and transketolase changed the concn. of several metabolites it did not increase NADPH concn. Wild-type strains and strains overexpressing any of the 3 genes were characterized in exponential and stationary phases of culture in minimal medium, with glucose as the C source and ammonium or nitrate as the N source and final cell density limiting substrate. Enzymes, intermediary metabolites, polyol pools (intra- and extracellular), organic acids, growth rates and rate constant of induction of acid production in post-exponential phase were measured. None of the modified strains had a changed growth rate. Partial least square regressions showed correlations between NADPH concn. and concn. of a total of 40 enzymes and metabolites. Thus, it was possible to predict the intracellular NADPH concn. from the concn. of enzymes, polyols and oxalate, and it is suggested that this prediction may be used in screening for high NADPH levels in engineered strains or mutants of other organisms.
机译：NADPH的低可用性限制了许多生物合成反应和生物转化。以增加细胞NADPH浓度为目的。和/或黑曲霉中通过戊糖磷酸途径的通量，克隆了编码葡萄糖6-磷酸脱氢酶（gsdA），6-磷酸葡萄糖酸脱氢酶（gndA）和转酮醇酶（tktA）的基因，并在单独的菌株中过表达。细胞内NADPH浓度由于6-磷酸葡糖酸脱氢酶过量生产了13倍，因此PPAR的含量增加了2-9倍。尽管葡萄糖6-磷酸脱氢酶和转酮醇酶的过量生产改变了浓度。的几种代谢物并没有增加NADPH浓度。野生型菌株和过表达3个基因中的任何一个的菌株的特征是在基本培养基中以指数和固定相培养，葡萄糖为C来源，铵或硝酸盐为N来源，并最终限制了细胞密度。测量了酶，中间代谢物，多元醇池（细胞内和细胞外），有机酸，生长速率和指数期后诱导产酸的速率常数。修饰菌株均未改变生长速率。偏最小二乘回归显示NADPH concn之间的相关性。和concn。总共40种酶和代谢产物。因此，可以预测细胞内NADPH浓度。来自concn。酶，多元醇和草酸盐的含量，建议将该预测结果用于筛选工程改造菌株或其他生物突变体中的高NADPH水平。

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