Enhanced selenocysteine biosynthesis for seleno-methylselenocysteine production in Bacillus subtilis

Yin, Xian; Zhou, Yu; Yang, Haizhen; Liao, Yong-Hong; Ma, Tengbo; Wang, Fenghuan

doi:10.1007/s00253-023-12482-8

Cited by 7 publications

(10 citation statements)

References 53 publications

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“…SeMCys synthesis requires SMT to methylate SeCys, and both SAM and MMet act as methyl donors. In a previous study, enhanced SAM formation facilitated SeMCys production at 18 μg/L in B. subtilis GBACB [ 10 ]. In the present study, MMet was synthesized as a methylation donor.…”

Section: Resultsmentioning

confidence: 99%

“…Ser is the precursor of both Cys and SeCys, and the overexpression of Cys insensitive SATp successfully improves SeCys production [ 10 ]. Based on the improvement in the expression levels of pathway enzymes, further knockout of sdaA may promote intracellular content of Ser, which is a substrate of SATp; the strain promoted SeMCys production by 2.5-fold.…”

Section: Discussionmentioning

confidence: 99%

“…Based on the over-expression of SMT and improved SAM production by expressing S -adenosylmethionine synthetase (SAM2) from S. cerevisiae , our previous work enhanced intracellular SeCys levels to optimize the production of SeMCys in Bacillus subtilis (Fig. 1 ), and the heterologous gene integration strain produced SeMCys at 18 μg/L [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

“…The formation of SeCys depends on the sulfur metabolic pathway and the corresponding metabolite is cysteine (Cys), which is crucial in cellular physiology because of the reactivity of its SH group [ 17 ]. Not only is the biosynthesis of Cys tightly controlled, but the degradation of this amino acid is also strong, leading to the inability of SeCys to continuously accumulate in large amounts intracellularly [ 10 ]. Therefore, it is important to investigate the SeCys degradation pathway.…”

Section: Introductionmentioning

confidence: 99%

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Optimized methyl donor and reduced precursor degradation pathway for seleno-methylselenocysteine production in Bacillus subtilis

Yin,

Zhao,

Zhou

et al. 2023

Microb Cell Fact

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Background Seleno-methylselenocysteine (SeMCys) is an effective component of selenium supplementation with anti-carcinogenic potential that can ameliorate neuropathology and cognitive deficits. In a previous study, a SeMCys producing strain of Bacillus subtilis GBACB was generated by releasing feedback inhibition by overexpression of cysteine-insensitive serine O-acetyltransferase, enhancing the synthesis of S-adenosylmethionine as methyl donor by overexpression of S-adenosylmethionine synthetase, and expressing heterologous selenocysteine methyltransferase. In this study, we aimed to improve GBACB SeMCys production by synthesizing methylmethionine as a donor to methylate selenocysteine and by inhibiting the precursor degradation pathway. Results First, the performance of three methionine S-methyltransferases that provide methylmethionine as a methyl donor for SeMCys production was determined. Integration of the NmMmt gene into GBACB improved SeMCys production from 20.7 to 687.4 μg/L. Next, the major routes for the degradation of selenocysteine, which is the precursor of SeMCys, were revealed by comparing selenocysteine hyper-accumulating and non-producing strains at the transcriptional level. The iscSB knockout strain doubled SeMCys production. Moreover, deleting sdaA, which is responsible for the degradation of serine as a precursor of selenocysteine, enhanced SeMCys production to 4120.3 μg/L. Finally, the culture conditions in the flasks were optimized. The strain was tolerant to higher selenite content in the liquid medium and the titer of SeMCys reached 7.5 mg/L. Conclusions The significance of methylmethionine as a methyl donor for SeMCys production in B. subtilis is reported, and enhanced precursor supply facilitates SeMCys synthesis. The results represent the highest SeMCys production to date and provide insight into Se metabolism.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Optimized methyl donor and reduced precursor degradation pathway for seleno-methylselenocysteine production in Bacillus subtilis

Yin,

Zhao,

Zhou

et al. 2023

Microb Cell Fact

Self Cite

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“…This was resolved by simply overexpressing the methionine synthase to increase the SAM level 3-fold . In other studies, direct overexpression of the SAM synthetases enhanced the intracellular SAM concentration and product titers. ,, In the production of methyl anthranilate, overexpression of homoserine succinyl-transferase and l -serine O -acetyltransferase which separately convert aspartate and serine into l -homocysteine increased the availability of the SAM level in the constructed SAM recycling loop and boosted the methyl anthranilate titer . This SAM regeneration cycle was then explored in the production of ferulic acid, where another two key enzymes encoding S -ribosylhomocysteine lyase and S -adenosylhomocysteine nucleosidase were overexpressed to increase the SAM level .…”

Section: Metabolic Regulation and Engineering Strategies Of Sam Biosy...mentioning

confidence: 99%

Improving Microbial Cell Factory Performance by Engineering SAM Availability

Lv,

Chang,

Zhang

et al. 2024

J. Agric. Food Chem.

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Methylated natural products are widely spread in nature. S-Adenosyl-L-methionine (SAM) is the secondary abundant cofactor and the primary methyl donor, which confer natural products with structural and functional diversification. The increasing demand for SAM-dependent natural products (SdNPs) has motivated the development of microbial cell factories (MCFs) for sustainable and efficient SdNP production. Insufficient and unsustainable SAM availability hinders the improvement of SdNP MCF performance. From the perspective of developing MCF, this review summarized recent understanding of de novo SAM biosynthesis and its regulatory mechanism. SAM is just the methyl mediator but not the original methyl source. Effective and sustainable methyl source supply is critical for efficient SdNP production. We compared and discussed the innate and relatively less explored alternative methyl sources and identified the one involving cheap one-carbon compound as more promising. The SAM biosynthesis is synergistically regulated on multilevels and is tightly connected with ATP and NAD(P)H pools. We also covered the recent advancement of metabolic engineering in improving intracellular SAM availability and SdNP production. Dynamic regulation is a promising strategy to achieve accurate and dynamic fine-tuning of intracellular SAM pool size. Finally, we discussed the design and engineering constraints underlying construction of SAM-responsive genetic circuits and envisioned their future applications in developing SdNP MCFs.

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Microbial Production of Amino Acids and Their Applications in Health and Nutrition Sectors

Agrawal,

Maharana,

Mohanty

et al. 2024

Microbial Products for Health and Nutrition

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Enhanced selenocysteine biosynthesis for seleno-methylselenocysteine production in Bacillus subtilis

Cited by 7 publications

References 53 publications

Optimized methyl donor and reduced precursor degradation pathway for seleno-methylselenocysteine production in Bacillus subtilis

Optimized methyl donor and reduced precursor degradation pathway for seleno-methylselenocysteine production in Bacillus subtilis

Improving Microbial Cell Factory Performance by Engineering SAM Availability

Microbial Production of Amino Acids and Their Applications in Health and Nutrition Sectors

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