Corrections to: Enhanced lincomycin production by co‑overexpression of metK1 and metK2 in Streptomyces lincolnensis

Xu, Yurong; Tan, Guoqing; Ke, Meilan; Li, Jie; Tang, Yulong; Meng, Sitong; Niu, Jingjing; Wang, Yansheng; Liu, Ruihua; Wu, Honglu; Bai, Linquan; Zhang, Lixin; Zhang, Buchang

doi:10.1007/s10295-018-2036-2

Cited by 3 publications

(4 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Escherichia coli strains were cultured at 37°C in Luria Bertani (LB) medium and supplemented with antibiotics as required (Sambrook and Russell 2012). S. lincolnensis strains were cultured at 30°C with shaking at 220 rpm in liquid TSBY medium for genomic DNA extraction (Xu et al 2018). S. lincolnensis strains were grown on MGM medium for spore production (Xu et al 2019).…”

Section: Strains Plasmids and Cultivation Conditionsmentioning

confidence: 99%

New Targets of TetR-type regulator SLCG_2919 for Controlling Lincomycin Biosynthesis in Streptomyces lincolnensis

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

The transcription factor (TF)-mediated regulatory network controlling lincomycin production in Streptomyces lincolnensis is yet to be fully elucidated despite several types of associated TFs having been reported. SLCG_2919, a tetracycline repressor (TetR)-type regulator, was the first TF to be characterized outside the lincomycin biosynthetic cluster to directly suppress the lincomycin biosynthesis in S. lincolnensis. In this study, improved genomic systematic evolution of ligands by exponential enrichment (gSELEX), an in-vitro technique, was adopted to capture additional SLCG_2919-targeted sequences harbouring the promoter regions of SLCG_6675, SLCG_4123-4124, SLCG_6579, and SLCG_0139-0140. These results were confirmed by electrophoretic mobility shift assays (EMSAs). RT-qPCR showed that the corresponding target genes SLCG_6675 (anthranilate synthase), SLCG_0139 (LysR family transcriptional regulator), SLCG_0140 (beta-lactamase), SLCG_6579 (cytochrome P450), SLCG_4123 (bifunctional DNA primase/polymerase), and SLCG_4124 (magnesium or magnesium-dependent protein phosphatase) in ΔSLCGL_2919 were differentially increased by 3.3-, 4.2-, 3.2-, 2.5-, 4.6-, and 2.2-fold relative to those in the parental strain S. lincolnensis LCGL. Furthermore, the individual inactivation of these target genes in LCGL reduced the lincomycin yield to varying degrees. This investigation expands on the known DNA targets of SLCG_2919 to control lincomycin production, and lays the foundation for improving industrial lincomycin yields via genetic engineering of this regulatory network.

show abstract

Section: Strains Plasmids and Cultivation Conditionsmentioning

confidence: 99%

New Targets of TetR-type regulator SLCG_2919 for Controlling Lincomycin Biosynthesis in Streptomyces lincolnensis

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Intracellular SAM was extracted and detected as described (Xu et al 2018). In brie y, 200 mg frozen cells were transferred to 1.5 mL microcentrifuge cubes and extracted with 1 mL of 1 mol/L formic acid at 4 °C for 2 h. The samples were centrifuged at 10000 rpm for 10 min at 4 °C to collect the supernatant for determination.…”

Section: Determination Of Intracellular Sam Concentrationmentioning

confidence: 99%

“…Given the high clinical importance of lincomycin, enhancement of lincomycin A production or reduction of lincomycin B production in S. lincolnensis have been performed in many studies by strain screening (Ye et al 2009;Meng et al 2013;Huang et al 2017), fermentation engineering (Xue et al 2009; Lee et al 2014; Zhuang et al 2019) or genetic manipulation (Pang et al 2015;Xu et al 2018). All the above studies focused on optimization of medium composition and process conditions or screening strains with more productivity.…”

Section: Introductionmentioning

confidence: 99%

Decreased lincomycin B content by regulating osmotic pressure in fermentation of Streptomyces lincolnensis

Zhang

Gao

2022

Preprint

View full text Add to dashboard Cite

Lincomycin, a clinically important antibiotic against gram-positive bacteria, is a sulfur-containing metabolite produced by Streptomyces lincolnensis. The content of lincomycin B is an important evaluation index of fermentation quality. In this study, we investigated the effects of NaCl as an osmotic regulator on lincomycin production. Based on the flask experiment, the optimal initial osmotic pressure for lincomycin fermentation was in the range between 0.860–1.355 Osmol/kg. The effect of osmotic pressure on the fermentation of lincomycin in 15 L bioreactor showed that the content of lincomycin B in the broth was decreased to 5.78% when adding 10 g/L NaCl while the content of lincomycin B was 8.20% in the control. During the fermentation process, the intracellular homocysteine and S-adenosylmethionine was higher when adding 10 g/L NaCl, which could provide more methyl donor for lincomycin production, causing the reduction of lincomycin B. Besides, high-throughput RNA sequencing was applied to study the mechanism of osmotic regulation on lincomycin fermentation. Under osmotic stimulation, the transcription of 23 genes in lincomycin biosynthesis cluster were significantly up-regulated at 41 h and the transcription of genes in HMP and EMP was up-regulated while the transcription of genes in TCA cycle were decreased. Notably, the genes involved in sulfur assimilation, mycothiol, ergothioneine and S-adenosylmethionine biosynthesis were up-regulated, which can provide more methyl donor (S-adenosylmethionine) and sulfur donor (mycothiol and ergothioneine) during lincomycin fermentation. This might be the reason of increased lincomycin A production and decreased lincomycin B content. Our study provides new insight into the mechanisms underlying osmotic pressure regulation on lincomycin fermentation and may be applied to improve secondary metabolite production.

show abstract

“…They promoted spiramycin production by increasing the concentration of ethylmalonyl-CoA via over-expressing the crotonyl coenzyme A reductase gene. S-Adenosylmethionine (SAM) is widely known to be a methyl donor to secondary metabolites [10,11]. Experimental results from Streptomyces lividans showed that overexpression of the metK gene that encodes SAM synthetase not only promotes higher methyl donor concentrations, but also stimulates the expression of a positive regulatory gene actII-ORF4 for the actinorhodin (Act) gene cluster, thereby leading to enhancement of Act biosynthesis in S. lividans [12,13].…”

Section: Introductionmentioning

confidence: 99%

Enhancement of toyocamycin production through increasing supply of precursor GTP in Streptomyces diastatochromogenes 1628

Zhang

et al. 2022

J Basic Microbiol

View full text Add to dashboard Cite

The nucleoside antibiotic toyocamycin (TM), which is produced by Streptomyces diastatochromogenes 1628, exhibits potent activity against a broad range of phytopathogenic fungi. TM was synthesized through a multi-step reaction, using guanosine triphosphate (GTP) as precursor. Based on a comparison of proteomics data from S. diastatochromogenes 1628 and rifamycin-resistant mutant 1628-T15 with high yield of TM, we determined that the differentially expressed protein X0NBV6 called ribose-phosphate pyrophosphokinase (RHP), which is a rate-limiting enzyme involved in the de novo biosynthesis of GTP, exhibits a higher expression level in mutant 1628-T15. In this study, to elucidate the relationships between RHP, GTP, and TM production, the gene rhp sd encoding RHP was cloned and overexpressed in S. diastatochromogenes strain 1628. The recombinant strain S. diastatochromogenes 1628-RHP exhibited better performance at the transcriptional level of the rhp sd gene, as well as RHP enzymatic activity, intracellular GTP concentration, and TM production, compared to S. diastatochromogenes 1628. Finally, the yield of TM produced by S. diastatochromogenes 1628-RHP (340.2 mg/L) was 133.3% higher than that produced by S. diastatochromogenes1628. Moreover, the transcriptional level of toy genes involved in TM biosynthesis was enhanced due to the overexpression of the rhp sd gene.

show abstract

Corrections to: Enhanced lincomycin production by co‑overexpression of metK1 and metK2 in Streptomyces lincolnensis

Abstract: In the online published article, row value “pIB139-metK1-metK2” in table 1 has been processed incorrectly. The correct table is given below

Cited by 3 publications

References 0 publications

New Targets of TetR-type regulator SLCG_2919 for Controlling Lincomycin Biosynthesis in Streptomyces lincolnensis

New Targets of TetR-type regulator SLCG_2919 for Controlling Lincomycin Biosynthesis in Streptomyces lincolnensis

Decreased lincomycin B content by regulating osmotic pressure in fermentation of Streptomyces lincolnensis

Enhancement of toyocamycin production through increasing supply of precursor GTP in Streptomyces diastatochromogenes 1628

Contact Info

Product

Resources

About