High Production of Nattokinase via Fed-Batch Fermentation of the γ-PGA-Deficient Strain of <i>Bacillus licheniformis</i>
During the production of nattokinase (NK) by <i>Bacillus</i> species, certain by-products such as poly-γ-glutamic acid (γ-PGA) are simultaneously synthesized. The impact of γ-PGA synthesis on NK production remains unclear. In this study, we knocked out the <i>pgsC</i> gene, a...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-12-01
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| Series: | Fermentation |
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| Online Access: | https://www.mdpi.com/2311-5637/9/12/1018 |
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| author | Xin Li Jing Yang Jun Liu Xiaohui Zhang Wei Wu Dazhong Yan Lihong Miao Dongbo Cai Xin Ma Shouwen Chen |
| author_facet | Xin Li Jing Yang Jun Liu Xiaohui Zhang Wei Wu Dazhong Yan Lihong Miao Dongbo Cai Xin Ma Shouwen Chen |
| author_sort | Xin Li |
| collection | DOAJ |
| description | During the production of nattokinase (NK) by <i>Bacillus</i> species, certain by-products such as poly-γ-glutamic acid (γ-PGA) are simultaneously synthesized. The impact of γ-PGA synthesis on NK production remains unclear. In this study, we knocked out the <i>pgsC</i> gene, a component of the γ-PGA synthetase cluster (<i>pgsBCA</i>), and constructed a novel recombinant strain, <i>Bacillus licheniformis</i> BL11. Next, we compared the fed-batch fermentation profiles of BL11 and its parental strain BL10, conducted transcriptional analysis, and measured intracellular ATP content. We also optimized glucose-feeding strategies under varying oxygen supply conditions. Our results indicated that the utilization rates of glucose and soybean meal were both improved in the <i>pgsC</i>-deficient strain BL11, and NK activity was enhanced. Furthermore, the transcriptional levels of genes involved in glycolysis and the TCA cycle were relatively upregulated in BL11. The maximal NK activity reached 2522.2 FU/mL at 54 h of fermentation using a constant glucose-feeding rate of 5.0 g/(L·h) under high oxygen supply conditions. The newly developed recombinant strain <i>B. licheniformis</i> BL11, along with the optimized feeding strategy, shows promise for large-scale NK production. |
| first_indexed | 2024-03-08T20:47:09Z |
| format | Article |
| id | doaj.art-cc42ca8b3fdb4ed58a15af6e5cf5a257 |
| institution | Directory Open Access Journal |
| issn | 2311-5637 |
| language | English |
| last_indexed | 2024-03-08T20:47:09Z |
| publishDate | 2023-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Fermentation |
| spelling | doaj.art-cc42ca8b3fdb4ed58a15af6e5cf5a2572023-12-22T14:07:55ZengMDPI AGFermentation2311-56372023-12-01912101810.3390/fermentation9121018High Production of Nattokinase via Fed-Batch Fermentation of the γ-PGA-Deficient Strain of <i>Bacillus licheniformis</i>Xin Li0Jing Yang1Jun Liu2Xiaohui Zhang3Wei Wu4Dazhong Yan5Lihong Miao6Dongbo Cai7Xin Ma8Shouwen Chen9School of Life Science and Technology, Wuhan Polytechnic University, Wuhan 430023, ChinaSchool of Life Science and Technology, Wuhan Polytechnic University, Wuhan 430023, ChinaSchool of Life Science and Technology, Wuhan Polytechnic University, Wuhan 430023, ChinaSchool of Life Science and Technology, Wuhan Polytechnic University, Wuhan 430023, ChinaSchool of Life Science and Technology, Wuhan Polytechnic University, Wuhan 430023, ChinaSchool of Life Science and Technology, Wuhan Polytechnic University, Wuhan 430023, ChinaSchool of Life Science and Technology, Wuhan Polytechnic University, Wuhan 430023, ChinaState Key Laboratory of Biocatalysis and Enzyme Engineering, Environmental Microbial Technology Center of Hubei Province, College of Life Sciences, Hubei University, Wuhan 430062, ChinaState Key Laboratory of Biocatalysis and Enzyme Engineering, Environmental Microbial Technology Center of Hubei Province, College of Life Sciences, Hubei University, Wuhan 430062, ChinaState Key Laboratory of Biocatalysis and Enzyme Engineering, Environmental Microbial Technology Center of Hubei Province, College of Life Sciences, Hubei University, Wuhan 430062, ChinaDuring the production of nattokinase (NK) by <i>Bacillus</i> species, certain by-products such as poly-γ-glutamic acid (γ-PGA) are simultaneously synthesized. The impact of γ-PGA synthesis on NK production remains unclear. In this study, we knocked out the <i>pgsC</i> gene, a component of the γ-PGA synthetase cluster (<i>pgsBCA</i>), and constructed a novel recombinant strain, <i>Bacillus licheniformis</i> BL11. Next, we compared the fed-batch fermentation profiles of BL11 and its parental strain BL10, conducted transcriptional analysis, and measured intracellular ATP content. We also optimized glucose-feeding strategies under varying oxygen supply conditions. Our results indicated that the utilization rates of glucose and soybean meal were both improved in the <i>pgsC</i>-deficient strain BL11, and NK activity was enhanced. Furthermore, the transcriptional levels of genes involved in glycolysis and the TCA cycle were relatively upregulated in BL11. The maximal NK activity reached 2522.2 FU/mL at 54 h of fermentation using a constant glucose-feeding rate of 5.0 g/(L·h) under high oxygen supply conditions. The newly developed recombinant strain <i>B. licheniformis</i> BL11, along with the optimized feeding strategy, shows promise for large-scale NK production.https://www.mdpi.com/2311-5637/9/12/1018nattokinase productionγ-PGA deficiency<i>Bacillus licheniformis</i>fed-batch fermentation |
| spellingShingle | Xin Li Jing Yang Jun Liu Xiaohui Zhang Wei Wu Dazhong Yan Lihong Miao Dongbo Cai Xin Ma Shouwen Chen High Production of Nattokinase via Fed-Batch Fermentation of the γ-PGA-Deficient Strain of <i>Bacillus licheniformis</i> Fermentation nattokinase production γ-PGA deficiency <i>Bacillus licheniformis</i> fed-batch fermentation |
| title | High Production of Nattokinase via Fed-Batch Fermentation of the γ-PGA-Deficient Strain of <i>Bacillus licheniformis</i> |
| title_full | High Production of Nattokinase via Fed-Batch Fermentation of the γ-PGA-Deficient Strain of <i>Bacillus licheniformis</i> |
| title_fullStr | High Production of Nattokinase via Fed-Batch Fermentation of the γ-PGA-Deficient Strain of <i>Bacillus licheniformis</i> |
| title_full_unstemmed | High Production of Nattokinase via Fed-Batch Fermentation of the γ-PGA-Deficient Strain of <i>Bacillus licheniformis</i> |
| title_short | High Production of Nattokinase via Fed-Batch Fermentation of the γ-PGA-Deficient Strain of <i>Bacillus licheniformis</i> |
| title_sort | high production of nattokinase via fed batch fermentation of the γ pga deficient strain of i bacillus licheniformis i |
| topic | nattokinase production γ-PGA deficiency <i>Bacillus licheniformis</i> fed-batch fermentation |
| url | https://www.mdpi.com/2311-5637/9/12/1018 |
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