Metabolic engineering of Acremonium chrysogenum for improving cephalosporin C production independent of methionine stimulation.
Metabolic engineering of Acremonium chrysogenum for improving cephalosporin C production independent of methionine stimulation.
复制标题
产黄顶孢霉的代谢工程可提高头孢菌素 C 的产量，且不依赖于蛋氨酸刺激
DOI：
10.1186/s12934-018-0936-5
复制
发表时间：
2018-06-07
影响因子：
6.4
通讯作者：
Liu G
中科院分区：
文献类型：
--
作者：
Liu J;Gao W;Pan Y;Liu G
由Acremium chrysogenum产生的头孢菌素C（CPC）是治疗细菌感染性疾病最重要的药物之一。蛋氨酸作为主要的兴奋剂，广泛应用于CPC的工业生产中。在这项研究中，我们发现蛋氨酸通过增强内源性 S-腺苷蛋氨酸 (SAM) 的积累来刺激 CPC 的产生。为了克服 CPC 产生的蛋氨酸依赖性刺激，通过代谢工程重建了 A. chrysogenum 的蛋氨酸循环。通过过表达 SAM 合成酶基因 AcsamS 和胱硫醚-γ-裂解酶基因 mecB，并分别破坏 SAM 依赖性甲基转移酶基因 Acppm1，获得了三个工程菌株。 AcsamS 的过度表达导致 CPC 产量增加四倍，达到 129.7 µg/mL。 Acppm1 的破坏还通过增强细胞内 SAM 的积累来增加 CPC 的产量（高达 135.5 µg/mL）。最后，通过在Acppm1破坏突变体中过表达mecB，构建了最佳重组菌株（Acppm1DM-mecBOE）。在该菌株中，CPC 产量达到最大值（142.7 µg/mL），是野生型水平的 5.5 倍，并且其提高完全独立于蛋氨酸刺激。在这项研究中，我们构建了一种重组菌株，其中 CPC 产量的提高完全独立于蛋氨酸刺激。这项工作为通过代谢工程提高A. chrysogenum CPC 产量提供了一条经济途径。本文的在线版本 (10.1186/s12934-018-0936-5) 包含补充材料，可供授权用户使用。
Cephalosporin C (CPC) produced by Acremonium chrysogenum is one of the most important drugs for treatment of bacterial infectious diseases. As the major stimulant, methionine is widely used in the industrial production of CPC. In this study, we found methionine stimulated CPC production through enhancing the accumulation of endogenous S-adenosylmethionine (SAM). To overcome the methionine dependent stimulation of CPC production, the methionine cycle of A. chrysogenum was reconstructed by metabolic engineering. Three engineered strains were obtained by overexpressing the SAM synthetase gene AcsamS and the cystathionine-γ-lyase gene mecB, and disrupting a SAM dependent methyltransferase gene Acppm1, respectively. Overexpression of AcsamS resulted in fourfold increase of CPC production which reached to 129.7 µg/mL. Disruption of Acppm1 also increased CPC production (up to 135.5 µg/mL) through enhancing the accumulation of intracellular SAM. Finally, an optimum recombinant strain (Acppm1DM-mecBOE) was constructed through overexpressing mecB in the Acppm1 disruption mutant. In this strain, CPC production reached to the maximum value (142.7 µg/mL) which was 5.5-fold of the wild-type level and its improvement was totally independent of methionine stimulation. In this study, we constructed a recombinant strain in which the improvement of CPC production was totally independent of methionine stimulation. This work provides an economic route for improving CPC production in A. chrysogenum through metabolic engineering. The online version of this article (10.1186/s12934-018-0936-5) contains supplementary material, which is available to authorized users.
登录
查看更多内容
影响因子：
4.8
作者：
Martínez-Chantar, ML;Latasa, MU;Avila, MA
通讯作者：
Avila, MA
影响因子：
33.6
作者：
Lu SC;Mato JM
通讯作者：
Mato JM
DOI：
10.1128/aem.10.4.321-325.1962
发表时间：
1962-01-01
期刊：
APPLIED MICROBIOLOGY
影响因子：
--
作者：
DEMAIN, AL;NEWKIRK, JF
通讯作者：
NEWKIRK, JF
影响因子：
9.8
作者：
MUDD, SH;POOLE, JR
通讯作者：
POOLE, JR
产黄顶孢霉中氮调节基因 AcareA 的破坏导致头孢菌素产量减少并抑制氮代谢
影响因子：
3
作者：
Li, Jinyang;Pan, Yuanyuan;Liu, Gang
通讯作者：
Liu, Gang