Engineering Bafilomycin High-Producers by Manipulating Regulatory and Biosynthetic Genes in the Marine Bacterium Streptomyces lohii.
Engineering Bafilomycin High-Producers by Manipulating Regulatory and Biosynthetic Genes in the Marine Bacterium Streptomyces lohii.
复制标题
DOI：
10.3390/md19010029
复制
发表时间：
2021-01-11
期刊：
影响因子：
5.4
通讯作者：
Li S
中科院分区：
文献类型：
--
作者：
Li Z;Li S;Du L;Zhang X;Jiang Y;Liu W;Zhang W;Li S
巴弗洛霉素A1是普卡环内酯天然产物家族的代表性化合物。这16元环plecomacrolide具有强大的抗真菌和液泡H+-ATP酶抑制活性。在我们的前期工作中，我们从海洋细菌Streptomyces lohii ATCC BAA-1276中鉴定了一个巴弗洛霉素生物合成基因簇（baf），其中基于生物信息学分析揭示了luxR家族调控基因orf 1和afsR家族调控基因bafG。在这项研究中，orf 1和bafG的巴夫洛霉素生物合成的正调控作用的特点是通过基因失活和过表达。与野生型S.在lohii菌株中，orf 1或bafG的敲除完全消除了巴非霉素的产生。orf 1或bafG的过表达分别导致巴非霉素的产量增加1.3倍和0.5倍。一个基因工程的S。在优化的摇瓶发酵培养基中，一株orf 1过表达、orf 2和orf 3失活的lohii菌株（SLO-08）单独产生巴夫洛霉素A1，效价为535.1 ± 25.0 mg/L。该重组菌在大规模生产新药巴夫洛霉素A1方面具有很大的应用潜力。
Bafilomycin A1 is the representative compound of the plecomacrolide natural product family. This 16-membered ring plecomacrolide has potent antifungal and vacuolar H+-ATPase inhibitory activities. In our previous work, we identified a bafilomycin biosynthetic gene cluster (baf) from the marine bacterium Streptomyces lohii ATCC BAA-1276, wherein a luxR family regulatory gene orf1 and an afsR family regulatory gene bafG were revealed based on bioinformatics analysis. In this study, the positive regulatory roles of orf1 and bafG for bafilomycin biosynthesis are characterized through gene inactivation and overexpression. Compared to the wild-type S. lohii strain, the knockout of either orf1 or bafG completely abolished the production of bafilomycins. The overexpression of orf1 or bafG led to 1.3- and 0.5-fold increased production of bafilomycins, respectively. A genetically engineered S. lohii strain (SLO-08) with orf1 overexpression and inactivation of the biosynthetic genes orf2 and orf3, solely produced bafilomycin A1 with the titer of 535.1 ± 25.0 mg/L in an optimized fermentation medium in shaking flasks. This recombinant strain holds considerable application potential in large-scale production of bafilomycin A1 for new drug development.
登录
查看更多内容
DOI：
10.1007/s10295-003-0063-z
发表时间：
2003-08-01
影响因子：
3.4
作者：
Horinouchi, S
通讯作者：
Horinouchi, S
DOI：
10.1073/pnas.88.24.11115
发表时间：
1991-12-01
影响因子：
11.1
作者：
CHOI, SH;GREENBERG, EP
通讯作者：
GREENBERG, EP
影响因子：
3.2
作者：
Choi, SU;Lee, CK;Nihira, T
通讯作者：
Nihira, T
影响因子：
16.6
作者：
Kleinbeck, Florian;Carreira, Erick M.
通讯作者：
Carreira, Erick M.
影响因子：
5.6
作者：
Ducros, VMA;Lewis, RJ;Brannigan, JA
通讯作者：
Brannigan, JA