Optimal Secretory Expression of Acetaldehyde Dehydrogenase from <i>Issatchenkia terricola</i> in <i>Bacillus subtilis</i> through a Combined Strategy
Acetaldehyde dehydrogenases are potential enzyme preparations that can be used to detoxify acetaldehyde and other exogenous aldehydes from pharmaceuticals, food, and biofuel production. In this study, we enhanced the expression of acetaldehyde dehydrogenase sourced from <i>Issatchenkia terrico...
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MDPI AG
2022-01-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/27/3/747 |
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| author | Jing Lu Yu Zhao Yu Cheng Rong Hu Yaowei Fang MingSheng Lyu Shujun Wang Zhaoxin Lu |
| author_facet | Jing Lu Yu Zhao Yu Cheng Rong Hu Yaowei Fang MingSheng Lyu Shujun Wang Zhaoxin Lu |
| author_sort | Jing Lu |
| collection | DOAJ |
| description | Acetaldehyde dehydrogenases are potential enzyme preparations that can be used to detoxify acetaldehyde and other exogenous aldehydes from pharmaceuticals, food, and biofuel production. In this study, we enhanced the expression of acetaldehyde dehydrogenase sourced from <i>Issatchenkia terricola</i> (istALDH) in <i>Bacillus subtilis</i> using a combinatorial strategy for the optimization of signal peptides, promoters, and growth conditions. First, a library of various signal peptides was constructed to identify the optimal signal peptides for efficient istALDH secretion. The signal peptide yqzG achieved the highest extracellular istALDH activity (204.85 ± 3.31 U/mL). Second, the aprE promoter was replaced by a constitutive promoter (i.e., P43) and an inducible promoter (i.e., Pglv), resulting in 12.40% and 19.97% enhanced istALDH, respectively. Furthermore, the tandem promoter P43-Pglv provided a better performance, resulting in 30.96% enhanced istALDH activity. Third, the production of istALDH was optimized by testing one factor at a time. Physical parameters were optimized including the inducer (e.g., maltose) concentrations, incubation temperatures, and inoculation amounts, and the results were 2.0%, 35 <inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><msup><mrow></mrow><mo>∘</mo></msup></semantics></math></inline-formula>C, and 2.0%, respectively. The optimized medium results were 2.0% glucose, 1.5% peptone, 2.5% yeast extract, 1% NaCl, and 0.5% (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub>. The extracellular istALDH activity was 331.19 ± 4.19 U/mL, yielding the highest production reported in the literature to date. |
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| series | Molecules |
| spelling | doaj.art-6ad3538837cf4dcfaef8dee44bd636942023-11-23T17:11:49ZengMDPI AGMolecules1420-30492022-01-0127374710.3390/molecules27030747Optimal Secretory Expression of Acetaldehyde Dehydrogenase from <i>Issatchenkia terricola</i> in <i>Bacillus subtilis</i> through a Combined StrategyJing Lu0Yu Zhao1Yu Cheng2Rong Hu3Yaowei Fang4MingSheng Lyu5Shujun Wang6Zhaoxin Lu7Jiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaCollege of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, ChinaAcetaldehyde dehydrogenases are potential enzyme preparations that can be used to detoxify acetaldehyde and other exogenous aldehydes from pharmaceuticals, food, and biofuel production. In this study, we enhanced the expression of acetaldehyde dehydrogenase sourced from <i>Issatchenkia terricola</i> (istALDH) in <i>Bacillus subtilis</i> using a combinatorial strategy for the optimization of signal peptides, promoters, and growth conditions. First, a library of various signal peptides was constructed to identify the optimal signal peptides for efficient istALDH secretion. The signal peptide yqzG achieved the highest extracellular istALDH activity (204.85 ± 3.31 U/mL). Second, the aprE promoter was replaced by a constitutive promoter (i.e., P43) and an inducible promoter (i.e., Pglv), resulting in 12.40% and 19.97% enhanced istALDH, respectively. Furthermore, the tandem promoter P43-Pglv provided a better performance, resulting in 30.96% enhanced istALDH activity. Third, the production of istALDH was optimized by testing one factor at a time. Physical parameters were optimized including the inducer (e.g., maltose) concentrations, incubation temperatures, and inoculation amounts, and the results were 2.0%, 35 <inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><msup><mrow></mrow><mo>∘</mo></msup></semantics></math></inline-formula>C, and 2.0%, respectively. The optimized medium results were 2.0% glucose, 1.5% peptone, 2.5% yeast extract, 1% NaCl, and 0.5% (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub>. The extracellular istALDH activity was 331.19 ± 4.19 U/mL, yielding the highest production reported in the literature to date.https://www.mdpi.com/1420-3049/27/3/747acetaldehyde dehydrogenase<i>Bacillus subtilis</i>signal peptidetandem promoteroptimization of fermentation |
| spellingShingle | Jing Lu Yu Zhao Yu Cheng Rong Hu Yaowei Fang MingSheng Lyu Shujun Wang Zhaoxin Lu Optimal Secretory Expression of Acetaldehyde Dehydrogenase from <i>Issatchenkia terricola</i> in <i>Bacillus subtilis</i> through a Combined Strategy Molecules acetaldehyde dehydrogenase <i>Bacillus subtilis</i> signal peptide tandem promoter optimization of fermentation |
| title | Optimal Secretory Expression of Acetaldehyde Dehydrogenase from <i>Issatchenkia terricola</i> in <i>Bacillus subtilis</i> through a Combined Strategy |
| title_full | Optimal Secretory Expression of Acetaldehyde Dehydrogenase from <i>Issatchenkia terricola</i> in <i>Bacillus subtilis</i> through a Combined Strategy |
| title_fullStr | Optimal Secretory Expression of Acetaldehyde Dehydrogenase from <i>Issatchenkia terricola</i> in <i>Bacillus subtilis</i> through a Combined Strategy |
| title_full_unstemmed | Optimal Secretory Expression of Acetaldehyde Dehydrogenase from <i>Issatchenkia terricola</i> in <i>Bacillus subtilis</i> through a Combined Strategy |
| title_short | Optimal Secretory Expression of Acetaldehyde Dehydrogenase from <i>Issatchenkia terricola</i> in <i>Bacillus subtilis</i> through a Combined Strategy |
| title_sort | optimal secretory expression of acetaldehyde dehydrogenase from i issatchenkia terricola i in i bacillus subtilis i through a combined strategy |
| topic | acetaldehyde dehydrogenase <i>Bacillus subtilis</i> signal peptide tandem promoter optimization of fermentation |
| url | https://www.mdpi.com/1420-3049/27/3/747 |
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