Fermentation Kinetics and Gene Expression Patterns in Adenosine Biosynthesis by <i>Bacillus subtilis</i>

Adenosine holds significant application value in the fields of food additives and pharmaceutical intermediate synthesis. Engineering strains to enhance their efficiency in utilizing fermentation substrates is considered an effective strategy for improving production yield. However, modifications to...

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Main Authors: Gengxuan Yan, Haoqiong Wu, Shumei Zhang, Chunyan Liu, Yuan Tian, Chong Yu
Format: Article
Language:English
Published: MDPI AG 2025-07-01
Series:Applied Sciences
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Online Access:https://www.mdpi.com/2076-3417/15/13/7592
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author Gengxuan Yan
Haoqiong Wu
Shumei Zhang
Chunyan Liu
Yuan Tian
Chong Yu
author_facet Gengxuan Yan
Haoqiong Wu
Shumei Zhang
Chunyan Liu
Yuan Tian
Chong Yu
author_sort Gengxuan Yan
collection DOAJ
description Adenosine holds significant application value in the fields of food additives and pharmaceutical intermediate synthesis. Engineering strains to enhance their efficiency in utilizing fermentation substrates is considered an effective strategy for improving production yield. However, modifications to adenosine-producing strains remain challenging due to the complex physiological and metabolic regulation governing adenosine biosynthesis. In this study, the molecular regulatory mechanisms of adenosine biosynthesis in a high-yielding <i>Bacillus subtilis</i> strain were analyzed through transcriptome sequencing. Under conditions in which an additional 10 g/L glutamine and 6 g/L hypoxanthine were supplemented at 48 h of cultivation to promote adenosine synthesis, a total of 105 significantly differentially expressed genes (69 downregulated and 36 upregulated) were identified, with key genes related to adenosine biosynthesis primarily concentrated in the downstream purine metabolic pathway. Notably, core biosynthetic genes including <i>purD</i>, <i>guaC</i>, <i>purH</i>, and <i>purN</i> showed significant downregulation in the high-yielding strain, suggesting that adenosine accumulation might inhibit related gene expression through negative feedback mechanisms. Fermentation kinetic analysis revealed that biomass reached its peak at 48 h (OD<sub>600</sub> = 0.82), with a glucose consumption rate of 73.28% at this stage. Gene expression pattern analysis demonstrated that <i>purD</i>, <i>guaC</i>, <i>purH</i>, and <i>purN</i> maintained relatively stable expression levels during fermentation. However, the exogenous supplementation of inosine (6 g/L) and glutamine (10 g/L) induced significant inhibition of their expression—a trend paralleling that observed with exogenous adenosine addition. This research elucidates key regulatory nodes in the adenosine biosynthesis of <i>Bacillus subtilis</i> and provides theoretical support and candidate targets for the targeted modification of industrial strains through metabolic engineering strategies.
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spelling doaj-art-59f423d3eb654312bc3e935ef4d9720a2025-08-20T03:50:16ZengMDPI AGApplied Sciences2076-34172025-07-011513759210.3390/app15137592Fermentation Kinetics and Gene Expression Patterns in Adenosine Biosynthesis by <i>Bacillus subtilis</i>Gengxuan Yan0Haoqiong Wu1Shumei Zhang2Chunyan Liu3Yuan Tian4Chong Yu5Institute of Microbiology Heilongjiang Academy of Sciences, Harbin 150010, ChinaInstitute of Microbiology Heilongjiang Academy of Sciences, Harbin 150010, ChinaInstitute of Microbiology Heilongjiang Academy of Sciences, Harbin 150010, ChinaInstitute of Microbiology Heilongjiang Academy of Sciences, Harbin 150010, ChinaInstitute of Microbiology Heilongjiang Academy of Sciences, Harbin 150010, ChinaInstitute of Microbiology Heilongjiang Academy of Sciences, Harbin 150010, ChinaAdenosine holds significant application value in the fields of food additives and pharmaceutical intermediate synthesis. Engineering strains to enhance their efficiency in utilizing fermentation substrates is considered an effective strategy for improving production yield. However, modifications to adenosine-producing strains remain challenging due to the complex physiological and metabolic regulation governing adenosine biosynthesis. In this study, the molecular regulatory mechanisms of adenosine biosynthesis in a high-yielding <i>Bacillus subtilis</i> strain were analyzed through transcriptome sequencing. Under conditions in which an additional 10 g/L glutamine and 6 g/L hypoxanthine were supplemented at 48 h of cultivation to promote adenosine synthesis, a total of 105 significantly differentially expressed genes (69 downregulated and 36 upregulated) were identified, with key genes related to adenosine biosynthesis primarily concentrated in the downstream purine metabolic pathway. Notably, core biosynthetic genes including <i>purD</i>, <i>guaC</i>, <i>purH</i>, and <i>purN</i> showed significant downregulation in the high-yielding strain, suggesting that adenosine accumulation might inhibit related gene expression through negative feedback mechanisms. Fermentation kinetic analysis revealed that biomass reached its peak at 48 h (OD<sub>600</sub> = 0.82), with a glucose consumption rate of 73.28% at this stage. Gene expression pattern analysis demonstrated that <i>purD</i>, <i>guaC</i>, <i>purH</i>, and <i>purN</i> maintained relatively stable expression levels during fermentation. However, the exogenous supplementation of inosine (6 g/L) and glutamine (10 g/L) induced significant inhibition of their expression—a trend paralleling that observed with exogenous adenosine addition. This research elucidates key regulatory nodes in the adenosine biosynthesis of <i>Bacillus subtilis</i> and provides theoretical support and candidate targets for the targeted modification of industrial strains through metabolic engineering strategies.https://www.mdpi.com/2076-3417/15/13/7592<i>Bacillus subtilis</i>fermentation kineticsadenosine biosynthesisgene expression
spellingShingle Gengxuan Yan
Haoqiong Wu
Shumei Zhang
Chunyan Liu
Yuan Tian
Chong Yu
Fermentation Kinetics and Gene Expression Patterns in Adenosine Biosynthesis by <i>Bacillus subtilis</i>
Applied Sciences
<i>Bacillus subtilis</i>
fermentation kinetics
adenosine biosynthesis
gene expression
title Fermentation Kinetics and Gene Expression Patterns in Adenosine Biosynthesis by <i>Bacillus subtilis</i>
title_full Fermentation Kinetics and Gene Expression Patterns in Adenosine Biosynthesis by <i>Bacillus subtilis</i>
title_fullStr Fermentation Kinetics and Gene Expression Patterns in Adenosine Biosynthesis by <i>Bacillus subtilis</i>
title_full_unstemmed Fermentation Kinetics and Gene Expression Patterns in Adenosine Biosynthesis by <i>Bacillus subtilis</i>
title_short Fermentation Kinetics and Gene Expression Patterns in Adenosine Biosynthesis by <i>Bacillus subtilis</i>
title_sort fermentation kinetics and gene expression patterns in adenosine biosynthesis by i bacillus subtilis i
topic <i>Bacillus subtilis</i>
fermentation kinetics
adenosine biosynthesis
gene expression
url https://www.mdpi.com/2076-3417/15/13/7592
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