Inactivation of SACE_3446, a TetR family transcriptional regulator, stimulates erythromycin production in Saccharopolyspora erythraea
Erythromycin A is a widely used antibiotic produced by Saccharopolyspora erythraea; however, its biosynthetic cluster lacks a regulatory gene, limiting the yield enhancement via regulation engineering of S. erythraea. Herein, six TetR family transcriptional regulators (TFRs) belonging to three genom...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
KeAi Communications Co., Ltd.
2016-03-01
|
| Series: | Synthetic and Systems Biotechnology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405805X15300077 |
| _version_ | 1797205324804915200 |
|---|---|
| author | Hang Wu Yansheng Wang Li Yuan Yongrong Mao Weiwei Wang Lin Zhu Panpan Wu Chengzhang Fu Rolf Müller David T. Weaver Lixin Zhang Buchang Zhang |
| author_facet | Hang Wu Yansheng Wang Li Yuan Yongrong Mao Weiwei Wang Lin Zhu Panpan Wu Chengzhang Fu Rolf Müller David T. Weaver Lixin Zhang Buchang Zhang |
| author_sort | Hang Wu |
| collection | DOAJ |
| description | Erythromycin A is a widely used antibiotic produced by Saccharopolyspora erythraea; however, its biosynthetic cluster lacks a regulatory gene, limiting the yield enhancement via regulation engineering of S. erythraea. Herein, six TetR family transcriptional regulators (TFRs) belonging to three genomic context types were individually inactivated in S. erythraea A226, and one of them, SACE_3446, was proved to play a negative role in regulating erythromycin biosynthesis. EMSA and qRT-PCR analysis revealed that SACE_3446 covering intact N-terminal DNA binding domain specifically bound to the promoter regions of erythromycin biosynthetic gene eryAI, the resistant gene ermE and the adjacent gene SACE_3447 (encoding a long-chain fatty-acid CoA ligase), and repressed their transcription. Furthermore, we explored the interaction relationships of SACE_3446 and previously identified TFRs (SACE_3986 and SACE_7301) associated with erythromycin production. Given demonstrated relatively independent regulation mode of SACE_3446 and SACE_3986 in erythromycin biosynthesis, we individually and concomitantly inactivated them in an industrial S. erythraea WB. Compared with WB, the WBΔ3446 and WBΔ3446Δ3986 mutants respectively displayed 36% and 65% yield enhancement of erythromycin A, following significantly elevated transcription of eryAI and ermE. When cultured in a 5 L fermentor, erythromycin A of WBΔ3446 and WBΔ3446Δ3986 successively reached 4095 mg/L and 4670 mg/L with 23% and 41% production improvement relative to WB. The strategy reported here will be useful to improve antibiotics production in other industrial actinomycete. |
| first_indexed | 2024-04-24T08:49:19Z |
| format | Article |
| id | doaj.art-e47d1d79d04c4605adcfdc4a2ca15204 |
| institution | Directory Open Access Journal |
| issn | 2405-805X |
| language | English |
| last_indexed | 2024-04-24T08:49:19Z |
| publishDate | 2016-03-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Synthetic and Systems Biotechnology |
| spelling | doaj.art-e47d1d79d04c4605adcfdc4a2ca152042024-04-16T12:56:11ZengKeAi Communications Co., Ltd.Synthetic and Systems Biotechnology2405-805X2016-03-0111394610.1016/j.synbio.2016.01.004Inactivation of SACE_3446, a TetR family transcriptional regulator, stimulates erythromycin production in Saccharopolyspora erythraeaHang Wu0Yansheng Wang1Li Yuan2Yongrong Mao3Weiwei Wang4Lin Zhu5Panpan Wu6Chengzhang Fu7Rolf Müller8David T. Weaver9Lixin Zhang10Buchang Zhang11Institute of Health Sciences, School of Life Sciences, Anhui University, Hefei 230601, ChinaInstitute of Health Sciences, School of Life Sciences, Anhui University, Hefei 230601, ChinaInstitute of Health Sciences, School of Life Sciences, Anhui University, Hefei 230601, ChinaInstitute of Health Sciences, School of Life Sciences, Anhui University, Hefei 230601, ChinaInstitute of Health Sciences, School of Life Sciences, Anhui University, Hefei 230601, ChinaInstitute of Health Sciences, School of Life Sciences, Anhui University, Hefei 230601, ChinaInstitute of Health Sciences, School of Life Sciences, Anhui University, Hefei 230601, ChinaCAS Key Laboratory of Pathogenic Microbiology & Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, ChinaHelmholtz Institute for Pharmaceutical Research, Helmholtz Centre for Infection Research and Department of Pharmaceutical Biotechnology, Saarland University, P.O. Box 15115, 66041 Saarbrücken, GermanyInstitute of Health Sciences, School of Life Sciences, Anhui University, Hefei 230601, ChinaInstitute of Health Sciences, School of Life Sciences, Anhui University, Hefei 230601, ChinaInstitute of Health Sciences, School of Life Sciences, Anhui University, Hefei 230601, ChinaErythromycin A is a widely used antibiotic produced by Saccharopolyspora erythraea; however, its biosynthetic cluster lacks a regulatory gene, limiting the yield enhancement via regulation engineering of S. erythraea. Herein, six TetR family transcriptional regulators (TFRs) belonging to three genomic context types were individually inactivated in S. erythraea A226, and one of them, SACE_3446, was proved to play a negative role in regulating erythromycin biosynthesis. EMSA and qRT-PCR analysis revealed that SACE_3446 covering intact N-terminal DNA binding domain specifically bound to the promoter regions of erythromycin biosynthetic gene eryAI, the resistant gene ermE and the adjacent gene SACE_3447 (encoding a long-chain fatty-acid CoA ligase), and repressed their transcription. Furthermore, we explored the interaction relationships of SACE_3446 and previously identified TFRs (SACE_3986 and SACE_7301) associated with erythromycin production. Given demonstrated relatively independent regulation mode of SACE_3446 and SACE_3986 in erythromycin biosynthesis, we individually and concomitantly inactivated them in an industrial S. erythraea WB. Compared with WB, the WBΔ3446 and WBΔ3446Δ3986 mutants respectively displayed 36% and 65% yield enhancement of erythromycin A, following significantly elevated transcription of eryAI and ermE. When cultured in a 5 L fermentor, erythromycin A of WBΔ3446 and WBΔ3446Δ3986 successively reached 4095 mg/L and 4670 mg/L with 23% and 41% production improvement relative to WB. The strategy reported here will be useful to improve antibiotics production in other industrial actinomycete.http://www.sciencedirect.com/science/article/pii/S2405805X15300077Saccharopolyspora erythraeaErythromycinTetR familySACE_3446Regulatory network |
| spellingShingle | Hang Wu Yansheng Wang Li Yuan Yongrong Mao Weiwei Wang Lin Zhu Panpan Wu Chengzhang Fu Rolf Müller David T. Weaver Lixin Zhang Buchang Zhang Inactivation of SACE_3446, a TetR family transcriptional regulator, stimulates erythromycin production in Saccharopolyspora erythraea Synthetic and Systems Biotechnology Saccharopolyspora erythraea Erythromycin TetR family SACE_3446 Regulatory network |
| title | Inactivation of SACE_3446, a TetR family transcriptional regulator, stimulates erythromycin production in Saccharopolyspora erythraea |
| title_full | Inactivation of SACE_3446, a TetR family transcriptional regulator, stimulates erythromycin production in Saccharopolyspora erythraea |
| title_fullStr | Inactivation of SACE_3446, a TetR family transcriptional regulator, stimulates erythromycin production in Saccharopolyspora erythraea |
| title_full_unstemmed | Inactivation of SACE_3446, a TetR family transcriptional regulator, stimulates erythromycin production in Saccharopolyspora erythraea |
| title_short | Inactivation of SACE_3446, a TetR family transcriptional regulator, stimulates erythromycin production in Saccharopolyspora erythraea |
| title_sort | inactivation of sace 3446 a tetr family transcriptional regulator stimulates erythromycin production in saccharopolyspora erythraea |
| topic | Saccharopolyspora erythraea Erythromycin TetR family SACE_3446 Regulatory network |
| url | http://www.sciencedirect.com/science/article/pii/S2405805X15300077 |
| work_keys_str_mv | AT hangwu inactivationofsace3446atetrfamilytranscriptionalregulatorstimulateserythromycinproductioninsaccharopolysporaerythraea AT yanshengwang inactivationofsace3446atetrfamilytranscriptionalregulatorstimulateserythromycinproductioninsaccharopolysporaerythraea AT liyuan inactivationofsace3446atetrfamilytranscriptionalregulatorstimulateserythromycinproductioninsaccharopolysporaerythraea AT yongrongmao inactivationofsace3446atetrfamilytranscriptionalregulatorstimulateserythromycinproductioninsaccharopolysporaerythraea AT weiweiwang inactivationofsace3446atetrfamilytranscriptionalregulatorstimulateserythromycinproductioninsaccharopolysporaerythraea AT linzhu inactivationofsace3446atetrfamilytranscriptionalregulatorstimulateserythromycinproductioninsaccharopolysporaerythraea AT panpanwu inactivationofsace3446atetrfamilytranscriptionalregulatorstimulateserythromycinproductioninsaccharopolysporaerythraea AT chengzhangfu inactivationofsace3446atetrfamilytranscriptionalregulatorstimulateserythromycinproductioninsaccharopolysporaerythraea AT rolfmuller inactivationofsace3446atetrfamilytranscriptionalregulatorstimulateserythromycinproductioninsaccharopolysporaerythraea AT davidtweaver inactivationofsace3446atetrfamilytranscriptionalregulatorstimulateserythromycinproductioninsaccharopolysporaerythraea AT lixinzhang inactivationofsace3446atetrfamilytranscriptionalregulatorstimulateserythromycinproductioninsaccharopolysporaerythraea AT buchangzhang inactivationofsace3446atetrfamilytranscriptionalregulatorstimulateserythromycinproductioninsaccharopolysporaerythraea |