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首页> 外文期刊>BMC Biotechnology >Enhanced itaconic acid production in Aspergillus niger using genetic modification and medium optimization
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Enhanced itaconic acid production in Aspergillus niger using genetic modification and medium optimization

机译：通过基因修饰和培养基优化提高黑曲霉中衣康酸的产量

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Background Aspergillus niger was selected as a host for producing itaconic acid due to its versatile and tolerant character in various growth environments, and its extremely high capacity of accumulating the precursor of itaconic acid: citric acid. Expressing the CAD gene from Aspergillus terreus opened the metabolic pathway towards itaconic acid in A. niger . In order to increase the production level, we continued by modifying its genome and optimizing cultivation media. Results Based on the results of previous transcriptomics studies and research from other groups, two genes : gpdA encoding the glyceraldehyde ?3-dehydrogenase (GPD) and hbd1 encoding a flavohemoglobin domain (HBD) were overexpressed in A. niger . Besides, new media were designed based on a reference medium for A. terreus . To analyze large numbers of cultures, we developed an approach for screening both fungal transformants and various media in 96-well micro-titer plates. The hbd1 transformants (HBD 2.2/2.5) did not improve itaconic acid titer while the gpdA transformant (GPD 4.3) decreased the itaconic acid production. Using 20 different media, copper was discovered to have a positive influence on itaconic acid production. Effects observed in the micro-titer plate screening were confirmed in controlled batch fermentation. Conclusions The performance of gpdA and hbd1 transformants was found not to be beneficial for itaconic acid production using the tested cultivation conditions. Medium optimization showed that, copper was positively correlated with improved itaconic acid production. Interestingly, the optimal conditions for itaconic acid clearly differ from conditions optimal for citric- and oxalic acid production.
机译：背景技术黑曲霉因其在各种生长环境中的通用性和耐受性，以及其积累衣康酸前体柠檬酸的极高能力而被选为生产衣康酸的宿主。从黑曲霉表达CAD基因开启了黑曲霉衣康酸的代谢途径。为了提高产量，我们继续修改其基因组并优化了培养基。结果基于先前的转录组学研究结果和其他研究的结果，两个基因：编码甘油醛β3-脱氢酶（GPD）的gpdA和编码黄素血红蛋白结构域（HBD）的hbd1在黑曲霉中过表达。此外，还根据土壤曲霉的参考培养基设计了新的培养基。为了分析大量的培养物，我们开发了一种在96孔微量滴定板中筛选真菌转化子和各种培养基的方法。 hbd1转化子（HBD 2.2 / 2.5）不能改善衣康酸的效价，而gpdA转化子（GPD 4.3）可以降低衣康酸的产生。发现使用20种不同的介质，铜对衣康酸的生产有积极影响。在微量滴定板筛选中观察到的效果在受控的分批发酵中得到确认。结论在所测试的培养条件下，发现gpdA和hbd1转化子的性能不利于衣康酸的生产。培养基优化表明，铜与衣康酸产量的提高呈正相关。有趣的是，衣康酸的最佳条件明显不同于柠檬酸和草酸生产的最佳条件。

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