Cis-Element Engineering Promotes the Expression of <i>Bacillus subtilis</i> Type I L-Asparaginase and Its Application in Food
Type I L-asparaginase from <i>Bacillus licheniformis</i> Z-1 (BlAase) was efficiently produced and secreted in <i>Bacillus subtilis</i> RIK 1285, but its low yield made it unsuitable for industrial use. Thus, a combined method was used in this study to boost BlAase synthesis...
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MDPI AG
2022-06-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/12/6588 |
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| author | Jiafeng Niu Ruxue Yan Juan Shen Xiaoyu Zhu Fanqiang Meng Zhaoxin Lu Fengxia Lu |
| author_facet | Jiafeng Niu Ruxue Yan Juan Shen Xiaoyu Zhu Fanqiang Meng Zhaoxin Lu Fengxia Lu |
| author_sort | Jiafeng Niu |
| collection | DOAJ |
| description | Type I L-asparaginase from <i>Bacillus licheniformis</i> Z-1 (BlAase) was efficiently produced and secreted in <i>Bacillus subtilis</i> RIK 1285, but its low yield made it unsuitable for industrial use. Thus, a combined method was used in this study to boost BlAase synthesis in <i>B. subtilis</i>. First, fifteen single strong promoters were chosen to replace the original promoter P43, with PyvyD achieving the greatest BlAase activity (436.28 U/mL). Second, dual-promoter systems were built using four promoters (PyvyD, P43, PaprE, and PspoVG) with relatively high BlAase expression levels to boost BlAase output, with the engine of promoter PaprE-PyvyD reaching 502.11 U/mL. The activity of BlAase was also increased (568.59 U/mL) by modifying key portions of the PaprE-PyvyD promoter. Third, when the ribosome binding site (RBS) sequence of promoter PyvyD was replaced, BlAase activity reached 790.1 U/mL, which was 2.27 times greater than the original promoter P43 strain. After 36 h of cultivation, the BlAase expression level in a 10 L fermenter reached 2163.09 U/mL, which was 6.2 times greater than the initial strain using promoter P43. Moreover, the application potential of BlAase on acrylamide migration in potato chips was evaluated. Results showed that 89.50% of acrylamide in fried potato chips could be removed when combined with blanching and BlAase treatment. These findings revealed that combining transcription and translation techniques are effective strategies to boost recombinant protein output, and BlAase can be a great candidate for controlling acrylamide in food processing. |
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| language | English |
| last_indexed | 2024-03-09T23:34:52Z |
| publishDate | 2022-06-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-2fd3b80bc93d4ecb9408593bbc065e582023-11-23T17:03:22ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-06-012312658810.3390/ijms23126588Cis-Element Engineering Promotes the Expression of <i>Bacillus subtilis</i> Type I L-Asparaginase and Its Application in FoodJiafeng Niu0Ruxue Yan1Juan Shen2Xiaoyu Zhu3Fanqiang Meng4Zhaoxin Lu5Fengxia Lu6College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, ChinaCollege of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, ChinaCollege of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, ChinaCollege of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, ChinaCollege of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, ChinaCollege of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, ChinaCollege of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, ChinaType I L-asparaginase from <i>Bacillus licheniformis</i> Z-1 (BlAase) was efficiently produced and secreted in <i>Bacillus subtilis</i> RIK 1285, but its low yield made it unsuitable for industrial use. Thus, a combined method was used in this study to boost BlAase synthesis in <i>B. subtilis</i>. First, fifteen single strong promoters were chosen to replace the original promoter P43, with PyvyD achieving the greatest BlAase activity (436.28 U/mL). Second, dual-promoter systems were built using four promoters (PyvyD, P43, PaprE, and PspoVG) with relatively high BlAase expression levels to boost BlAase output, with the engine of promoter PaprE-PyvyD reaching 502.11 U/mL. The activity of BlAase was also increased (568.59 U/mL) by modifying key portions of the PaprE-PyvyD promoter. Third, when the ribosome binding site (RBS) sequence of promoter PyvyD was replaced, BlAase activity reached 790.1 U/mL, which was 2.27 times greater than the original promoter P43 strain. After 36 h of cultivation, the BlAase expression level in a 10 L fermenter reached 2163.09 U/mL, which was 6.2 times greater than the initial strain using promoter P43. Moreover, the application potential of BlAase on acrylamide migration in potato chips was evaluated. Results showed that 89.50% of acrylamide in fried potato chips could be removed when combined with blanching and BlAase treatment. These findings revealed that combining transcription and translation techniques are effective strategies to boost recombinant protein output, and BlAase can be a great candidate for controlling acrylamide in food processing.https://www.mdpi.com/1422-0067/23/12/6588L-asparaginase<i>Bacillus subtilis</i>promoterdual-promoter systemsribosome binding sitescale-up fermentation |
| spellingShingle | Jiafeng Niu Ruxue Yan Juan Shen Xiaoyu Zhu Fanqiang Meng Zhaoxin Lu Fengxia Lu Cis-Element Engineering Promotes the Expression of <i>Bacillus subtilis</i> Type I L-Asparaginase and Its Application in Food International Journal of Molecular Sciences L-asparaginase <i>Bacillus subtilis</i> promoter dual-promoter systems ribosome binding site scale-up fermentation |
| title | Cis-Element Engineering Promotes the Expression of <i>Bacillus subtilis</i> Type I L-Asparaginase and Its Application in Food |
| title_full | Cis-Element Engineering Promotes the Expression of <i>Bacillus subtilis</i> Type I L-Asparaginase and Its Application in Food |
| title_fullStr | Cis-Element Engineering Promotes the Expression of <i>Bacillus subtilis</i> Type I L-Asparaginase and Its Application in Food |
| title_full_unstemmed | Cis-Element Engineering Promotes the Expression of <i>Bacillus subtilis</i> Type I L-Asparaginase and Its Application in Food |
| title_short | Cis-Element Engineering Promotes the Expression of <i>Bacillus subtilis</i> Type I L-Asparaginase and Its Application in Food |
| title_sort | cis element engineering promotes the expression of i bacillus subtilis i type i l asparaginase and its application in food |
| topic | L-asparaginase <i>Bacillus subtilis</i> promoter dual-promoter systems ribosome binding site scale-up fermentation |
| url | https://www.mdpi.com/1422-0067/23/12/6588 |
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