Construction and heterologous expression of the di-AFN A1 biosynthetic gene cluster in Streptomyces model strains.
10.1016/S1875-5364(22)60197-3
- Author:
Weijia WEI
1
,
2
;
Wenzhao WANG
3
;
Chao LI
1
,
2
;
Yue TANG
3
;
Zhengyan GUO
4
;
Yihua CHEN
1
,
5
Author Information
1. State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China
2. College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100149, China.
3. State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
4. Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing 100050, China.
5. College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100149, China. Electronic address: chenyihua@im.ac.cn.
- Publication Type:Journal Article
- Keywords:
Cyclohexapeptide;
Heterologous expression;
Streptomyces host;
Titer improvement;
di-AFN A(1)
- MeSH:
Cloning, Molecular;
Streptomyces/metabolism*;
Multigene Family;
Anti-Bacterial Agents/metabolism*;
Plasmids/genetics*
- From:
Chinese Journal of Natural Medicines (English Ed.)
2022;20(11):873-880
- CountryChina
- Language:English
-
Abstract:
Natural cyclohexapeptide AFN A1 fromStreptomyces alboflavus 313 has moderate antibacterial and antitumor activities. An artificial designed AFN A1 homodimer, di-AFN A1, is an antibiotic exhibiting 10 to 150 fold higher biological activities, compared with the monomer. Unfortunately, the yield of di-AFN A1 is very low (0.09 ± 0.03 mg·L-1) in the engineered strain Streptomyces alboflavus 313_hmtS (S. albo/313_hmtS), which is not friendly to be genetically engineered for titer improvement of di-AFN A1 production. In this study, we constructed a biosynthetic gene cluster for di-AFN A1 and increased its production through heterologous expression. During the collection of di-AFN A1 biosynthetic genes, the afn genes were located at three sites of S. alboflavus 313 genome. The di-AFN A1 biosynthetic gene cluster (BGC) was first assembled on one plasmid and introduced into the model strain Streptomyces lividans TK24, which produced di-AFN A1 at a titer of 0.43 ± 0.01 mg·L-1. To further increase the yield of di-AFN A1, the di-AFN A1 BGC was multiplied and split to mimic the natural afn biosynthetic genes, and the production of di-AFN A1 increased to 0.62 ± 0.11 mg·L-1 in S. lividans TK24 by the later strategy. Finally, different Streptomyces hosts were tested and the titer of di-AFN A1 increased to 0.81 ± 0.17 mg·L-1, about 8.0-fold higher than that in S. albo/313_hmtS. Successful heterologous expression of di-AFN A1 with a remarkable increased titer will greatly facilitate the following synthetic biological study and drug development of this dimeric cyclohexapeptide.
