Levansucrase from <i>Bacillus amyloliquefaciens</i> KK9 and Its Y237S Variant Producing the High Bioactive Levan-Type Fructooligosaccharides
Levan-typed fructooligosaccharide (LFOS), a β-2,6 linked oligofructose, displays the potential application as a prebiotic and therapeutic dietary supplement. In the present study, LFOS was synthesized using levansucrase from <i>Bacillus amyloliquefaciens</i> KK9 (LsKK9). The wild-type Ls...
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MDPI AG
2020-04-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/10/5/692 |
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| author | Pongsakorn Phengnoi Thanapon Charoenwongpaiboon Karan Wangpaiboon Methus Klaewkla Santhana Nakapong Wonnop Visessanguan Kazuo Ito Rath Pichyangkura Kamontip Kuttiyawong |
| author_facet | Pongsakorn Phengnoi Thanapon Charoenwongpaiboon Karan Wangpaiboon Methus Klaewkla Santhana Nakapong Wonnop Visessanguan Kazuo Ito Rath Pichyangkura Kamontip Kuttiyawong |
| author_sort | Pongsakorn Phengnoi |
| collection | DOAJ |
| description | Levan-typed fructooligosaccharide (LFOS), a β-2,6 linked oligofructose, displays the potential application as a prebiotic and therapeutic dietary supplement. In the present study, LFOS was synthesized using levansucrase from <i>Bacillus amyloliquefaciens</i> KK9 (LsKK9). The wild-type LsKK9 was cloned and expressed in <i>E. coli</i>, and purified by cation exchanger chromatography. Additionally, Y237S variant of LsKK9 was constructed based on sequence alignment and structural analysis to enhance the LFOS production. High-performance anion-exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD) analysis indicated that Y237S variant efficiently produced a higher amount of short-chain LFOS than wild type. Also, the concentration of enzyme and sucrose in the reactions was optimized. Finally, prebiotic activity assay demonstrated that LFOS produced by Y237S variant had higher prebiotic activity than that of the wild-type enzyme, making the variant enzyme attractive for food biotechnology. |
| first_indexed | 2024-03-10T20:09:21Z |
| format | Article |
| id | doaj.art-4b4e904cde954adb81a971184981b1ec |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-10T20:09:21Z |
| publishDate | 2020-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-4b4e904cde954adb81a971184981b1ec2023-11-19T23:03:51ZengMDPI AGBiomolecules2218-273X2020-04-0110569210.3390/biom10050692Levansucrase from <i>Bacillus amyloliquefaciens</i> KK9 and Its Y237S Variant Producing the High Bioactive Levan-Type FructooligosaccharidesPongsakorn Phengnoi0Thanapon Charoenwongpaiboon1Karan Wangpaiboon2Methus Klaewkla3Santhana Nakapong4Wonnop Visessanguan5Kazuo Ito6Rath Pichyangkura7Kamontip Kuttiyawong8Department of Chemistry, Faculty of Liberal Arts and Science, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom 73140, ThailandDepartment of Chemistry, Faculty of Science, Silpakorn University, Nakhon Pathom 73000, ThailandDepartment of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, ThailandDepartment of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, ThailandDepartment of Chemistry, Faculty of Science, Ramkhamhaeng University, Bangkok 10240, ThailandNational Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Pathumthani 12120, ThailandGraduate School of Science, Osaka City University, Osaka 558-8585, JapanDepartment of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, ThailandDepartment of Chemistry, Faculty of Liberal Arts and Science, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom 73140, ThailandLevan-typed fructooligosaccharide (LFOS), a β-2,6 linked oligofructose, displays the potential application as a prebiotic and therapeutic dietary supplement. In the present study, LFOS was synthesized using levansucrase from <i>Bacillus amyloliquefaciens</i> KK9 (LsKK9). The wild-type LsKK9 was cloned and expressed in <i>E. coli</i>, and purified by cation exchanger chromatography. Additionally, Y237S variant of LsKK9 was constructed based on sequence alignment and structural analysis to enhance the LFOS production. High-performance anion-exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD) analysis indicated that Y237S variant efficiently produced a higher amount of short-chain LFOS than wild type. Also, the concentration of enzyme and sucrose in the reactions was optimized. Finally, prebiotic activity assay demonstrated that LFOS produced by Y237S variant had higher prebiotic activity than that of the wild-type enzyme, making the variant enzyme attractive for food biotechnology.https://www.mdpi.com/2218-273X/10/5/692<i>Bacillus amyloliquefaciens</i>fructooligosaccharidelevansucraseprebioticmutagenesis |
| spellingShingle | Pongsakorn Phengnoi Thanapon Charoenwongpaiboon Karan Wangpaiboon Methus Klaewkla Santhana Nakapong Wonnop Visessanguan Kazuo Ito Rath Pichyangkura Kamontip Kuttiyawong Levansucrase from <i>Bacillus amyloliquefaciens</i> KK9 and Its Y237S Variant Producing the High Bioactive Levan-Type Fructooligosaccharides Biomolecules <i>Bacillus amyloliquefaciens</i> fructooligosaccharide levansucrase prebiotic mutagenesis |
| title | Levansucrase from <i>Bacillus amyloliquefaciens</i> KK9 and Its Y237S Variant Producing the High Bioactive Levan-Type Fructooligosaccharides |
| title_full | Levansucrase from <i>Bacillus amyloliquefaciens</i> KK9 and Its Y237S Variant Producing the High Bioactive Levan-Type Fructooligosaccharides |
| title_fullStr | Levansucrase from <i>Bacillus amyloliquefaciens</i> KK9 and Its Y237S Variant Producing the High Bioactive Levan-Type Fructooligosaccharides |
| title_full_unstemmed | Levansucrase from <i>Bacillus amyloliquefaciens</i> KK9 and Its Y237S Variant Producing the High Bioactive Levan-Type Fructooligosaccharides |
| title_short | Levansucrase from <i>Bacillus amyloliquefaciens</i> KK9 and Its Y237S Variant Producing the High Bioactive Levan-Type Fructooligosaccharides |
| title_sort | levansucrase from i bacillus amyloliquefaciens i kk9 and its y237s variant producing the high bioactive levan type fructooligosaccharides |
| topic | <i>Bacillus amyloliquefaciens</i> fructooligosaccharide levansucrase prebiotic mutagenesis |
| url | https://www.mdpi.com/2218-273X/10/5/692 |
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