Enhancing cellulase biosynthesis of Bacillus subtilis Z2 by regulating intracellular NADH level
Summary: Optimizing cellulase biosynthesis in Bacillus subtilis is crucial for enhancing enzymatic yield in lignocellulosic biomass conversion. However, the regulatory mechanisms linking intracellular NAD(H/+) levels to cellulase production remain elusive. In this study, we systematically screened 1...
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Elsevier
2025-05-01
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| Series: | iScience |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589004225006029 |
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| author | Shuai Liu Yi Li Lin Quan Hai-Xia Liu Yang Luo Yong-Zhong Wang |
| author_facet | Shuai Liu Yi Li Lin Quan Hai-Xia Liu Yang Luo Yong-Zhong Wang |
| author_sort | Shuai Liu |
| collection | DOAJ |
| description | Summary: Optimizing cellulase biosynthesis in Bacillus subtilis is crucial for enhancing enzymatic yield in lignocellulosic biomass conversion. However, the regulatory mechanisms linking intracellular NAD(H/+) levels to cellulase production remain elusive. In this study, we systematically screened 13 genes associated with NAD+ biosynthesis and NADH regeneration in B. subtilis Z2. Employing a modular engineering strategy with four distinct modules, we directed metabolic flux to enhance NAD+ biosynthesis and NADH regeneration. Key genes (ycel, nadV, nadM, mdh, and sucB) were identified, and their co-expression in B. subtilis Z2 resulted in a 13.09-fold increase in intracellular NADH levels and a consequential 2.24- and 2.04-fold enhancement in the filter paper-hydrolyzing (FPase [representing total cellulase]) activity and carboxymethylcellulose (CMCase [representing endoglucanase]) activity, respectively. Experimental validations, including antagonist LaCl3 treatment and spcF gene deletion, unequivocally established the calcium signaling pathway’s role in regulating cellulase gene overexpression in response to elevated intracellular NAD(H/+) levels. |
| format | Article |
| id | doaj-art-c5675913a39e4364a7d51ccdcc856e17 |
| institution | OA Journals |
| issn | 2589-0042 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj-art-c5675913a39e4364a7d51ccdcc856e172025-08-20T02:17:34ZengElsevieriScience2589-00422025-05-0128511234110.1016/j.isci.2025.112341Enhancing cellulase biosynthesis of Bacillus subtilis Z2 by regulating intracellular NADH levelShuai Liu0Yi Li1Lin Quan2Hai-Xia Liu3Yang Luo4Yong-Zhong Wang5Key Laboratory of Biorheological Science and Technology (Chongqing University), Ministry of Education, College of Bioengineering, Chongqing University, Chongqing 400030, ChinaKey Laboratory of Biorheological Science and Technology (Chongqing University), Ministry of Education, College of Bioengineering, Chongqing University, Chongqing 400030, ChinaKey Laboratory of Biorheological Science and Technology (Chongqing University), Ministry of Education, College of Bioengineering, Chongqing University, Chongqing 400030, ChinaChongqing Key Laboratory of Translational Research for Cancer Metastasis and Individualized Treatment, Chongqing University Cancer Hospital, Chongqing 400030, China; Corresponding authorCenter of Smart Laboratory and Molecular Medicine, NHC Key Laboratory of Birth Defects and Reproductive Health, Jiangjin Hospital, School of Medicine, Chongqing University, Chongqing 400030, China; Corresponding authorKey Laboratory of Biorheological Science and Technology (Chongqing University), Ministry of Education, College of Bioengineering, Chongqing University, Chongqing 400030, China; Corresponding authorSummary: Optimizing cellulase biosynthesis in Bacillus subtilis is crucial for enhancing enzymatic yield in lignocellulosic biomass conversion. However, the regulatory mechanisms linking intracellular NAD(H/+) levels to cellulase production remain elusive. In this study, we systematically screened 13 genes associated with NAD+ biosynthesis and NADH regeneration in B. subtilis Z2. Employing a modular engineering strategy with four distinct modules, we directed metabolic flux to enhance NAD+ biosynthesis and NADH regeneration. Key genes (ycel, nadV, nadM, mdh, and sucB) were identified, and their co-expression in B. subtilis Z2 resulted in a 13.09-fold increase in intracellular NADH levels and a consequential 2.24- and 2.04-fold enhancement in the filter paper-hydrolyzing (FPase [representing total cellulase]) activity and carboxymethylcellulose (CMCase [representing endoglucanase]) activity, respectively. Experimental validations, including antagonist LaCl3 treatment and spcF gene deletion, unequivocally established the calcium signaling pathway’s role in regulating cellulase gene overexpression in response to elevated intracellular NAD(H/+) levels.http://www.sciencedirect.com/science/article/pii/S2589004225006029Natural sciencesBiological sciencesBiotechnologyMicrobial biotechnologyEnzyme engineering |
| spellingShingle | Shuai Liu Yi Li Lin Quan Hai-Xia Liu Yang Luo Yong-Zhong Wang Enhancing cellulase biosynthesis of Bacillus subtilis Z2 by regulating intracellular NADH level iScience Natural sciences Biological sciences Biotechnology Microbial biotechnology Enzyme engineering |
| title | Enhancing cellulase biosynthesis of Bacillus subtilis Z2 by regulating intracellular NADH level |
| title_full | Enhancing cellulase biosynthesis of Bacillus subtilis Z2 by regulating intracellular NADH level |
| title_fullStr | Enhancing cellulase biosynthesis of Bacillus subtilis Z2 by regulating intracellular NADH level |
| title_full_unstemmed | Enhancing cellulase biosynthesis of Bacillus subtilis Z2 by regulating intracellular NADH level |
| title_short | Enhancing cellulase biosynthesis of Bacillus subtilis Z2 by regulating intracellular NADH level |
| title_sort | enhancing cellulase biosynthesis of bacillus subtilis z2 by regulating intracellular nadh level |
| topic | Natural sciences Biological sciences Biotechnology Microbial biotechnology Enzyme engineering |
| url | http://www.sciencedirect.com/science/article/pii/S2589004225006029 |
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