Property Improvement of α-Amylase from Bacillus stearothermophilus by Deletion of Amino Acid Residues Arginine 179 and Glycine 180
To improve the properties of α-amylase from Bacillus stearothermophilus (AmyS), a deletion mutant AmyS∆R179-G180 was constructed by deleting arginine (Arg179) and glycine (Gly180) using site-directed mutagenesis. AmyS and AmyS∆R179-G180 were expressed in Bacillus subtilis and purified by ammonium su...
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| Format: | Article |
| Language: | English |
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University of Zagreb Faculty of Food Technology and Biotechnology
2018-01-01
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| Series: | Food Technology and Biotechnology |
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| Online Access: | https://hrcak.srce.hr/file/290729 |
| _version_ | 1827282867333365760 |
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| author | Yuanming Gai Jingqi Chen Shibin Zhang Beiwei Zhu Dawei Zhang |
| author_facet | Yuanming Gai Jingqi Chen Shibin Zhang Beiwei Zhu Dawei Zhang |
| author_sort | Yuanming Gai |
| collection | DOAJ |
| description | To improve the properties of α-amylase from Bacillus stearothermophilus (AmyS), a deletion mutant AmyS∆R179-G180 was constructed by deleting arginine (Arg179) and glycine (Gly180) using site-directed mutagenesis. AmyS and AmyS∆R179-G180 were expressed in Bacillus subtilis and purified by ammonium sulfate precipitation, after which the enzymatic properties were characterized and compared. By deleting amino acids Arg179 and Gly180, the thermostability of α-amylase AmyS∆R179-G180 was enhanced and the half-life at 100 °C significantly increased from 24 to 33 min. In addition, AmyS∆R179-G180 exhibited greater acid resistance and lower calcium requirements to maintain α-amylase activity. The secretory capacity of the recombinant strain was evaluated by fed-batch fermentation in a 7.5-litre fermentor in which high α-amylase activity was obtained. The highest activity reached 3300 U/mL with a high productivity of 45.8 U/(mL•h). |
| first_indexed | 2024-04-24T09:26:19Z |
| format | Article |
| id | doaj.art-00ece8d37cb1458aa8d8ed4746e8e34f |
| institution | Directory Open Access Journal |
| issn | 1330-9862 1334-2606 |
| language | English |
| last_indexed | 2024-04-24T09:26:19Z |
| publishDate | 2018-01-01 |
| publisher | University of Zagreb Faculty of Food Technology and Biotechnology |
| record_format | Article |
| series | Food Technology and Biotechnology |
| spelling | doaj.art-00ece8d37cb1458aa8d8ed4746e8e34f2024-04-15T14:42:00ZengUniversity of Zagreb Faculty of Food Technology and BiotechnologyFood Technology and Biotechnology1330-98621334-26062018-01-01561586410.17113/ftb.56.01.18.5448Property Improvement of α-Amylase from Bacillus stearothermophilus by Deletion of Amino Acid Residues Arginine 179 and Glycine 180Yuanming Gai0Jingqi Chen1Shibin Zhang2Beiwei Zhu3Dawei Zhang4Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesTianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesKey Laboratory of Systems Microbial Biotechnology, Chinese Academy of SciencesSchool of Food Science and Technology, Dalian Polytechnic University, National Engineering Research Center of SeafoodKey Laboratory of Systems Microbial Biotechnology, Chinese Academy of SciencesTo improve the properties of α-amylase from Bacillus stearothermophilus (AmyS), a deletion mutant AmyS∆R179-G180 was constructed by deleting arginine (Arg179) and glycine (Gly180) using site-directed mutagenesis. AmyS and AmyS∆R179-G180 were expressed in Bacillus subtilis and purified by ammonium sulfate precipitation, after which the enzymatic properties were characterized and compared. By deleting amino acids Arg179 and Gly180, the thermostability of α-amylase AmyS∆R179-G180 was enhanced and the half-life at 100 °C significantly increased from 24 to 33 min. In addition, AmyS∆R179-G180 exhibited greater acid resistance and lower calcium requirements to maintain α-amylase activity. The secretory capacity of the recombinant strain was evaluated by fed-batch fermentation in a 7.5-litre fermentor in which high α-amylase activity was obtained. The highest activity reached 3300 U/mL with a high productivity of 45.8 U/(mL•h).https://hrcak.srce.hr/file/290729α-amylaseBacillus subtilisfermentationsite-directed mutagenesisthermostability |
| spellingShingle | Yuanming Gai Jingqi Chen Shibin Zhang Beiwei Zhu Dawei Zhang Property Improvement of α-Amylase from Bacillus stearothermophilus by Deletion of Amino Acid Residues Arginine 179 and Glycine 180 Food Technology and Biotechnology α-amylase Bacillus subtilis fermentation site-directed mutagenesis thermostability |
| title | Property Improvement of α-Amylase from Bacillus stearothermophilus by Deletion of Amino Acid Residues Arginine 179 and Glycine 180 |
| title_full | Property Improvement of α-Amylase from Bacillus stearothermophilus by Deletion of Amino Acid Residues Arginine 179 and Glycine 180 |
| title_fullStr | Property Improvement of α-Amylase from Bacillus stearothermophilus by Deletion of Amino Acid Residues Arginine 179 and Glycine 180 |
| title_full_unstemmed | Property Improvement of α-Amylase from Bacillus stearothermophilus by Deletion of Amino Acid Residues Arginine 179 and Glycine 180 |
| title_short | Property Improvement of α-Amylase from Bacillus stearothermophilus by Deletion of Amino Acid Residues Arginine 179 and Glycine 180 |
| title_sort | property improvement of α amylase from bacillus stearothermophilus by deletion of amino acid residues arginine 179 and glycine 180 |
| topic | α-amylase Bacillus subtilis fermentation site-directed mutagenesis thermostability |
| url | https://hrcak.srce.hr/file/290729 |
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