Enhancing trehalose production via Bacillus species G1 cyclodextrin glucanotransferase mutants: modifying disproportionation characteristics and thermal stability
Inefficient conversion of small molecule maltooligosaccharides into trehalose greatly affects the cost of the production of trehalose by double enzyme method [maltooligosyl trehalose synthase (MTSase) and maltooligosyl trehalose trehalohyrolase (MTHase)]. This study used directed evolution to increa...
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Frontiers Media S.A.
2024-11-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1500232/full |
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| author | Bobo Miao Bobo Miao Di Huang Di Huang Tengfei Wang Tengfei Wang Hongling Liu Hongling Liu Zhifeng Hao Haibo Yuan Haibo Yuan Yi Jiang Yi Jiang |
| author_facet | Bobo Miao Bobo Miao Di Huang Di Huang Tengfei Wang Tengfei Wang Hongling Liu Hongling Liu Zhifeng Hao Haibo Yuan Haibo Yuan Yi Jiang Yi Jiang |
| author_sort | Bobo Miao |
| collection | DOAJ |
| description | Inefficient conversion of small molecule maltooligosaccharides into trehalose greatly affects the cost of the production of trehalose by double enzyme method [maltooligosyl trehalose synthase (MTSase) and maltooligosyl trehalose trehalohyrolase (MTHase)]. This study used directed evolution to increase oligosaccharide utilization by the cyclomaltodextrin glucanotransferase (CGTase) from Bacillus species G1. This enzyme was chosen for its adaptability and stability in trehalose production. Model analysis revealed that the hydrogen bond distance between the N33K mutant and maltose reduced from 2.6 Å to 2.3 Å, increasing maltose affinity and boosting transglycosylation activity by 2.1-fold compared to the wild type. Further mutations improved thermal stability and optimum temperature, resulting in the N33K/S211G mutant. Consistent results from repeated experiments showed that the N33K/S211G mutant increased trehalose yield by 32.6% using maltodextrin. The results enhanced the double-enzyme method formed by MTSase and MTHase for trehalose production. Overall, we have identified optimal catalytic conditions, demonstrating significant potential for industrial-scale trehalose production with enhanced efficiency and cost-effectiveness. |
| format | Article |
| id | doaj-art-5f5b8bae82c14a9ba6b0beeab2911fdc |
| institution | Kabale University |
| issn | 1664-302X |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj-art-5f5b8bae82c14a9ba6b0beeab2911fdc2024-11-19T06:15:17ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2024-11-011510.3389/fmicb.2024.15002321500232Enhancing trehalose production via Bacillus species G1 cyclodextrin glucanotransferase mutants: modifying disproportionation characteristics and thermal stabilityBobo Miao0Bobo Miao1Di Huang2Di Huang3Tengfei Wang4Tengfei Wang5Hongling Liu6Hongling Liu7Zhifeng Hao8Haibo Yuan9Haibo Yuan10Yi Jiang11Yi Jiang12State Key Laboratory of Biobased Material and Green Papermaking (LBMP), Qilu University of Technology (Shandong Academy of Sciences), Jinan, Shandong, ChinaKey Laboratory of Shandong Microbial Engineering, College of Bioengineering, QiLu University of Technology (Shandong Academy of Sciences), Jinan, Shandong, ChinaState Key Laboratory of Biobased Material and Green Papermaking (LBMP), Qilu University of Technology (Shandong Academy of Sciences), Jinan, Shandong, ChinaKey Laboratory of Shandong Microbial Engineering, College of Bioengineering, QiLu University of Technology (Shandong Academy of Sciences), Jinan, Shandong, ChinaState Key Laboratory of Biobased Material and Green Papermaking (LBMP), Qilu University of Technology (Shandong Academy of Sciences), Jinan, Shandong, ChinaKey Laboratory of Shandong Microbial Engineering, College of Bioengineering, QiLu University of Technology (Shandong Academy of Sciences), Jinan, Shandong, ChinaState Key Laboratory of Biobased Material and Green Papermaking (LBMP), Qilu University of Technology (Shandong Academy of Sciences), Jinan, Shandong, ChinaKey Laboratory of Shandong Microbial Engineering, College of Bioengineering, QiLu University of Technology (Shandong Academy of Sciences), Jinan, Shandong, ChinaYantai Zhaoyi Biotechnology Co., Ltd, Yantai, ChinaState Key Laboratory of Biobased Material and Green Papermaking (LBMP), Qilu University of Technology (Shandong Academy of Sciences), Jinan, Shandong, ChinaKey Laboratory of Shandong Microbial Engineering, College of Bioengineering, QiLu University of Technology (Shandong Academy of Sciences), Jinan, Shandong, ChinaState Key Laboratory of Biobased Material and Green Papermaking (LBMP), Qilu University of Technology (Shandong Academy of Sciences), Jinan, Shandong, ChinaKey Laboratory of Shandong Microbial Engineering, College of Bioengineering, QiLu University of Technology (Shandong Academy of Sciences), Jinan, Shandong, ChinaInefficient conversion of small molecule maltooligosaccharides into trehalose greatly affects the cost of the production of trehalose by double enzyme method [maltooligosyl trehalose synthase (MTSase) and maltooligosyl trehalose trehalohyrolase (MTHase)]. This study used directed evolution to increase oligosaccharide utilization by the cyclomaltodextrin glucanotransferase (CGTase) from Bacillus species G1. This enzyme was chosen for its adaptability and stability in trehalose production. Model analysis revealed that the hydrogen bond distance between the N33K mutant and maltose reduced from 2.6 Å to 2.3 Å, increasing maltose affinity and boosting transglycosylation activity by 2.1-fold compared to the wild type. Further mutations improved thermal stability and optimum temperature, resulting in the N33K/S211G mutant. Consistent results from repeated experiments showed that the N33K/S211G mutant increased trehalose yield by 32.6% using maltodextrin. The results enhanced the double-enzyme method formed by MTSase and MTHase for trehalose production. Overall, we have identified optimal catalytic conditions, demonstrating significant potential for industrial-scale trehalose production with enhanced efficiency and cost-effectiveness.https://www.frontiersin.org/articles/10.3389/fmicb.2024.1500232/fulldirected evolutionmaltodextrinmaltoseprotein engineeringtransglycosylation |
| spellingShingle | Bobo Miao Bobo Miao Di Huang Di Huang Tengfei Wang Tengfei Wang Hongling Liu Hongling Liu Zhifeng Hao Haibo Yuan Haibo Yuan Yi Jiang Yi Jiang Enhancing trehalose production via Bacillus species G1 cyclodextrin glucanotransferase mutants: modifying disproportionation characteristics and thermal stability Frontiers in Microbiology directed evolution maltodextrin maltose protein engineering transglycosylation |
| title | Enhancing trehalose production via Bacillus species G1 cyclodextrin glucanotransferase mutants: modifying disproportionation characteristics and thermal stability |
| title_full | Enhancing trehalose production via Bacillus species G1 cyclodextrin glucanotransferase mutants: modifying disproportionation characteristics and thermal stability |
| title_fullStr | Enhancing trehalose production via Bacillus species G1 cyclodextrin glucanotransferase mutants: modifying disproportionation characteristics and thermal stability |
| title_full_unstemmed | Enhancing trehalose production via Bacillus species G1 cyclodextrin glucanotransferase mutants: modifying disproportionation characteristics and thermal stability |
| title_short | Enhancing trehalose production via Bacillus species G1 cyclodextrin glucanotransferase mutants: modifying disproportionation characteristics and thermal stability |
| title_sort | enhancing trehalose production via bacillus species g1 cyclodextrin glucanotransferase mutants modifying disproportionation characteristics and thermal stability |
| topic | directed evolution maltodextrin maltose protein engineering transglycosylation |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1500232/full |
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