Rapid improvement in the macrolactins production of Bacillus sp. combining atmospheric room temperature plasma with the specific growth rate index

Yi, Xiangxi; Gan, Yuman; Jiang, Lei; Yu, Lian; Liu, Yonghong; Gao, Chenghai

doi:10.1016/j.jbiosc.2020.02.014

Cited by 6 publications

(9 citation statements)

References 38 publications

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“…In this study, ALE was applied to develop a mutant strain for improving MLNs production. A72 showing excellent MLNs metabolic performance was selected as the parental strain for ALE [ 30 ]. Prior to initiating a random mutant experiment, i.e., ALE, an appropriate screening strategy is a contributing factor that determines whether the whole experiment is a success or not.…”

Section: Resultsmentioning

confidence: 99%

“…To the best of our knowledge, this is the first report on creating an excellent antibiotic production strain by using ALE with saline as the screening pressure. Yi et al applied ARTP mutant technique to develop a robust MLNs-producing strain[ 30 ]. According to the change in colony morphology after ARTP treatment, an excellent mutant strain was obtained with a low-screening efficiency of 6.67%.…”

Section: Discussionmentioning

confidence: 99%

“…A72 is a MLNs-producer strain [ 30 ], which was used as the parental strain for ALE. International Streptomyces Project (ISP) medium 2 (ISP-2) (number represents glucose concentration) containing 2 g/L yeast extract, 2 g/L malt extract, 3% (w/v) sea salt, and 2 g/L glucose at pH 6.5, was used for the cultivation of seed cells.…”

Section: Methodsmentioning

confidence: 99%

“…The glucose concentration in the fermentation broth was measured by a biosensor analyzer (Institute of Microbiology SBA-40E, Shangdong, China). The method for measuring MLNs production was described previously [ 30 ] with some modifications. Briefly, 1 mL of fermentation broth was added to 1.5 mL of methanol.…”

Section: Methodsmentioning

confidence: 99%

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Improvement of macrolactins production by the genetic adaptation of Bacillus siamensis A72 to saline stress via adaptive laboratory evolution

et al. 2022

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Background Macrolactins, a type of macrolide antibiotic, are toxic to the producer strains. As such, its level is usually maintained below the lethal concentration during the fermentation process. To improve the production of macrolactins, we applied adaptive laboratory evolution technology to engineer a saline-resistant mutant strain. The hypothesis that strains with saline resistance show improved macrolactins production was investigated. Results Using saline stress as a selective pressure, we engineered a mutant strain with saline resistance coupled with enhanced macrolactins production within 60 days using a self-made device. As compared with the parental strain, the evolved strain produced macrolactins with 11.93% improvement in non-saline stress fermentation medium containing 50 g/L glucose, when the glucose concentration increased to 70 g/L, the evolved strain produced macrolactins with 71.04% improvement. RNA sequencing and metabolomics results revealed that amino acid metabolism was involved in the production of macrolactins in the evolved strain. Furthermore, genome sequencing of the evolved strain revealed a candidate mutation, hisDD41Y, that was causal for the improved MLNs production, it was 3.42 times higher than the control in the overexpression hisDD41Y strain. Results revealed that saline resistance protected the producer strain from feedback inhibition of end-product (macrolide antibiotic), resulting in enhanced MLNs production. Conclusions In the present work, we successfully engineered a mutant strain with enhanced macrolactins production by adaptive laboratory evolution using saline stress as a selective pressure. Based on physiological, transcriptomic and genetic analysis, amino acid metabolism was found to benefit macrolactins production improvement. Our strategy might be applicable to improve the production of other kinds of macrolide antibiotics and other toxic compounds. The identification of the hisD mutation will allow for the deduction of metabolic engineering strategies in future research.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Improvement of macrolactins production by the genetic adaptation of Bacillus siamensis A72 to saline stress via adaptive laboratory evolution

et al. 2022

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“…Their common point was the conversion of glycine to arginine, which may be a strategy to improve polyketides. Yi et al (Yi et al 2020) obtained the high-yielding Bacillus sp. A29 and A72 through ARTP, and their macroactins production was 35.2% and 52.8% higher than the parent strain, respectively.…”

Section: Discussionmentioning

confidence: 99%

Random compound mutagenesis breeding of Bacillus amyloliquefaciens X030 with high resistance to pathogenic bacteria on grass carp

Zhou

Huang

et al. 2021

Preprint

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Background: Bacillus amyloliquefaciens X030 (BaX030) was obtained by screening from peanut fields in Henan Province, China. It had broad-spectrum antibacterial activity against fish pathogens Aeromonas hydrophila and Aeromonas veronii. In order to improve its antibacterial effect, BaX030 was carried out the compound mutagenesis that atmospheric and room temperature plasma (ARTP) combined with nitrosoguanidine (NTG). Results: The result showed the yield of macrolactin A and oxydifficidin of the mutant N-11 were increased by 2.01 times and 3.68 times, respectively. Re-sequencing found that the corresponding 9th and 15th gene clusters had 4 and 6 SNP mutations, respectively, the 15th gene cluster also had 7 InDel mutations. Scanning electron microscopy observed that the N-11 became thin and long. The results of qRT-PCR indicated that feeding the N-11 can increase the expression of immune factors in the liver or kidney tissue of grass carp. It can also significantly reduce the mortality and the surface symptoms of grass carp that was infected by two pathogens through HE staining and protection experiments.Conclusion: This is the first report that after ARTP-NTG compound mutagenesis, a high-yielding strain of macrolactin A and oxydifficidin were increased by 2.01 times and 3.68 times compared with the original strain, respectively, which laid the foundation for elucidating its biological regulation, they had antibacterial effects on Aeromonas hydrophila and Aeromonas veronii. We combined re-sequencing to find the mutation sites of gene clusters and. The probiotic strain N-11 can quickly activate the immune protection mechanism of grass carp to resist pathogenic bacteria.

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High-efficiency breeding of Bacillus siamensis with hyper macrolactins production using physical mutagenesis and a high-throughput culture system

Zhang,

Jiang,

Liu

et al. 2024

Journal of Biotechnology

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Rapid improvement in the macrolactins production of Bacillus sp. combining atmospheric room temperature plasma with the specific growth rate index

Cited by 6 publications

References 38 publications

Improvement of macrolactins production by the genetic adaptation of Bacillus siamensis A72 to saline stress via adaptive laboratory evolution

Improvement of macrolactins production by the genetic adaptation of Bacillus siamensis A72 to saline stress via adaptive laboratory evolution

Random compound mutagenesis breeding of Bacillus amyloliquefaciens X030 with high resistance to pathogenic bacteria on grass carp

High-efficiency breeding of Bacillus siamensis with hyper macrolactins production using physical mutagenesis and a high-throughput culture system

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