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Insights into the mechanism of enhanced tetramethylpyrazine production in dehulled adlay fermented by Bacillus subtilis BJ3-2.

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Τίτλος: Insights into the mechanism of enhanced tetramethylpyrazine production in dehulled adlay fermented by Bacillus subtilis BJ3-2.
Συγγραφείς: Ma Y; School of Liquor and Food Engineering, Guizhou University, Guiyang, 550025, China., Zhang H; Sinopharm Group of Tongjitang (Guizhou) Pharmaceutical Co., Ltd.,, Guiyang, 550025, China., Zhang R; Sinopharm Group of Tongjitang (Guizhou) Pharmaceutical Co., Ltd.,, Guiyang, 550025, China., Chen Q; Sinopharm Group of Tongjitang (Guizhou) Pharmaceutical Co., Ltd.,, Guiyang, 550025, China., Peng L; School of Liquor and Food Engineering, Guizhou University, Guiyang, 550025, China., Zeng H; School of Liquor and Food Engineering, Guizhou University, Guiyang, 550025, China., Qin L; School of Liquor and Food Engineering, Guizhou University, Guiyang, 550025, China; Guizhou Technology Innovation Center of Suantan, Anshun, 561000, China., Wu Y; Key Laboratory of Plant Resource Conservationand Germplasm Innovationin Mountainous Region (Ministry of Education), College of Life Sciences/Institute of Agro-Bioengineering, Guizhou University, Guiyang, 550025, China., Wen A; School of Liquor and Food Engineering, Guizhou University, Guiyang, 550025, China; Guizhou Technology Innovation Center of Suantan, Anshun, 561000, China. Electronic address: aywen@gzu.edu.cn., Sun Y; Sinopharm Group of Tongjitang (Guizhou) Pharmaceutical Co., Ltd.,, Guiyang, 550025, China. Electronic address: suntjt@126.com.
Πηγή: Food research international (Ottawa, Ont.) [Food Res Int] 2026 Oct 31; Vol. 242 (Pt 3), pp. 120080. Date of Electronic Publication: 2026 Jul 18.
Τύπος έκδοσης: Journal Article
Γλώσσα: English
Στοιχεία περιοδικού: Publisher: Published on behalf of the Canadian Institute of Food Science and Technology by Elsevier Applied Science Country of Publication: Canada NLM ID: 9210143 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-7145 (Electronic) Linking ISSN: 09639969 NLM ISO Abbreviation: Food Res Int Subsets: MEDLINE
Imprint Name(s): Original Publication: Ottawa, Ontario, Canada : Published on behalf of the Canadian Institute of Food Science and Technology by Elsevier Applied Science, c1992-
Ιατρικοί όροι (MeSH): Bacillus subtilis*/metabolism , Bacillus subtilis*/genetics , Pyrazines*/metabolism , Fermentation*, Bacterial Proteins/metabolism ; Bacterial Proteins/genetics ; Acetoin/metabolism ; Amino Acids/metabolism ; Proteomics ; Glycine max ; Gene Expression Regulation, Bacterial ; Molecular Docking Simulation
Περίληψη: Tetramethylpyrazine (TTMP) is a vital bioactive alkaloid and characteristic flavor compound in fermented foods. Our previous study found that fermentation of adlay by Bacillus subtilis BJ3-2 efficiently accumulates TTMP, whereas the underlying high-yield mechanism remains unclear. This study investigated the fermentation characteristics, gene transcription and protein expression of B. subtilis BJ3-2 in dehulled adlay (BDA) and soybean (BSB), respectively, and elucidated the mechanism responsible for high-yield TTMP production. The results showed that glutamate, leucine and phenylalanine were major free amino acids in BDA. The TTMP yield in BDA at 48 h (6.11 mg/g dry weight) was 360-fold higher than that in BSB. Transcriptomic and proteomic analysis demonstrated that compared with the soybean substrate, dehulled adlay substrate significantly up-regulated the expression of alsSD and ilvBH genes and their encoding proteins in B. subtilis BJ3-2, which were involved in C5-branched dibasic acid metabolism, 2-oxocarboxylic acid metabolism, and valine, leucine and isoleucine biosynthesis. Meanwhile, acetoin degradation was inhibited by down-regulating acetoin dehydrogenase complex (acoABCL) in citrate cycle, glycolysis/gluconeogenesis and carbon metabolism. Additionally, nitrogen metabolism pathway was transcriptionally enhanced to guarantee sufficient ammonium supply. Notably, protein-protein interaction and molecular docking analyses revealed that acetohydroxyacid synthase (ilvBH) interacted tightly with α-acetolactate decarboxylase (alsD), potentially forming a metabolic channel for acetoin synthesis. In conclusion, the efficient synthesis of TTMP in BDA was primarily attributed to the high synthesis and low degradation of acetoin, and the moderate synthesis of ammonium/ammonia. This study provided a theoretical basis for the targeted and efficient biosynthesis of TTMP.
(Copyright © 2026 Elsevier Ltd. All rights reserved.)
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Contributed Indexing: Keywords: Adlay; Bacillus subtilis BJ3–2; Proteomics; Tetramethylpyrazine; Transcriptomics
Substance Nomenclature: 0 (Pyrazines)
V80F4IA5XG (tetramethylpyrazine)
0 (Bacterial Proteins)
BG4D34CO2H (Acetoin)
0 (Amino Acids)
Entry Date(s): Date Created: 20260901 Date Completed: 20260901 Latest Revision: 20260901
Update Code: 20260902
DOI: 10.1016/j.foodres.2026.120080
PMID: 42680358
Βάση Δεδομένων: MEDLINE
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  Data: Insights into the mechanism of enhanced tetramethylpyrazine production in dehulled adlay fermented by Bacillus subtilis BJ3-2.
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  Data: <searchLink fieldCode="AU" term="%22Ma+Y%22">Ma Y</searchLink>; School of Liquor and Food Engineering, Guizhou University, Guiyang, 550025, China.<br /><searchLink fieldCode="AU" term="%22Zhang+H%22">Zhang H</searchLink>; Sinopharm Group of Tongjitang (Guizhou) Pharmaceutical Co., Ltd.,, Guiyang, 550025, China.<br /><searchLink fieldCode="AU" term="%22Zhang+R%22">Zhang R</searchLink>; Sinopharm Group of Tongjitang (Guizhou) Pharmaceutical Co., Ltd.,, Guiyang, 550025, China.<br /><searchLink fieldCode="AU" term="%22Chen+Q%22">Chen Q</searchLink>; Sinopharm Group of Tongjitang (Guizhou) Pharmaceutical Co., Ltd.,, Guiyang, 550025, China.<br /><searchLink fieldCode="AU" term="%22Peng+L%22">Peng L</searchLink>; School of Liquor and Food Engineering, Guizhou University, Guiyang, 550025, China.<br /><searchLink fieldCode="AU" term="%22Zeng+H%22">Zeng H</searchLink>; School of Liquor and Food Engineering, Guizhou University, Guiyang, 550025, China.<br /><searchLink fieldCode="AU" term="%22Qin+L%22">Qin L</searchLink>; School of Liquor and Food Engineering, Guizhou University, Guiyang, 550025, China; Guizhou Technology Innovation Center of Suantan, Anshun, 561000, China.<br /><searchLink fieldCode="AU" term="%22Wu+Y%22">Wu Y</searchLink>; Key Laboratory of Plant Resource Conservationand Germplasm Innovationin Mountainous Region (Ministry of Education), College of Life Sciences/Institute of Agro-Bioengineering, Guizhou University, Guiyang, 550025, China.<br /><searchLink fieldCode="AU" term="%22Wen+A%22">Wen A</searchLink>; School of Liquor and Food Engineering, Guizhou University, Guiyang, 550025, China; Guizhou Technology Innovation Center of Suantan, Anshun, 561000, China. Electronic address: aywen@gzu.edu.cn.<br /><searchLink fieldCode="AU" term="%22Sun+Y%22">Sun Y</searchLink>; Sinopharm Group of Tongjitang (Guizhou) Pharmaceutical Co., Ltd.,, Guiyang, 550025, China. Electronic address: suntjt@126.com.
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  Data: Tetramethylpyrazine (TTMP) is a vital bioactive alkaloid and characteristic flavor compound in fermented foods. Our previous study found that fermentation of adlay by Bacillus subtilis BJ3-2 efficiently accumulates TTMP, whereas the underlying high-yield mechanism remains unclear. This study investigated the fermentation characteristics, gene transcription and protein expression of B. subtilis BJ3-2 in dehulled adlay (BDA) and soybean (BSB), respectively, and elucidated the mechanism responsible for high-yield TTMP production. The results showed that glutamate, leucine and phenylalanine were major free amino acids in BDA. The TTMP yield in BDA at 48 h (6.11 mg/g dry weight) was 360-fold higher than that in BSB. Transcriptomic and proteomic analysis demonstrated that compared with the soybean substrate, dehulled adlay substrate significantly up-regulated the expression of alsSD and ilvBH genes and their encoding proteins in B. subtilis BJ3-2, which were involved in C5-branched dibasic acid metabolism, 2-oxocarboxylic acid metabolism, and valine, leucine and isoleucine biosynthesis. Meanwhile, acetoin degradation was inhibited by down-regulating acetoin dehydrogenase complex (acoABCL) in citrate cycle, glycolysis/gluconeogenesis and carbon metabolism. Additionally, nitrogen metabolism pathway was transcriptionally enhanced to guarantee sufficient ammonium supply. Notably, protein-protein interaction and molecular docking analyses revealed that acetohydroxyacid synthase (ilvBH) interacted tightly with α-acetolactate decarboxylase (alsD), potentially forming a metabolic channel for acetoin synthesis. In conclusion, the efficient synthesis of TTMP in BDA was primarily attributed to the high synthesis and low degradation of acetoin, and the moderate synthesis of ammonium/ammonia. This study provided a theoretical basis for the targeted and efficient biosynthesis of TTMP.<br /> (Copyright © 2026 Elsevier Ltd. All rights reserved.)
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  Data: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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  Data: <i>Keywords: </i>Adlay; Bacillus subtilis BJ3–2; Proteomics; Tetramethylpyrazine; Transcriptomics
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  Data: 0 (Pyrazines)<br />V80F4IA5XG (tetramethylpyrazine)<br />0 (Bacterial Proteins)<br />BG4D34CO2H (Acetoin)<br />0 (Amino Acids)
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        Type: general
      – SubjectFull: Bacterial Proteins genetics
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