Improved production of recombinant β-mannanase (TaMan5) in Pichia pastoris and its synergistic degradation of lignocellulosic biomass
Mannan, a highly abundant and cost-effective natural resource, holds great potential for the generation of high-value compounds such as bioactive polysaccharides and biofuels. In this study, we successfully enhanced the expression of constructed GH5 β-mannanase (TaMan5) from Trichoderma asperellum N...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2023-09-01
|
| Series: | Frontiers in Bioengineering and Biotechnology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2023.1244772/full |
| _version_ | 1797690433861582848 |
|---|---|
| author | Fengzhen Zheng Abdul Basit Zhiyue Zhang Huan Zhuang Jun Chen Jianfen Zhang |
| author_facet | Fengzhen Zheng Abdul Basit Zhiyue Zhang Huan Zhuang Jun Chen Jianfen Zhang |
| author_sort | Fengzhen Zheng |
| collection | DOAJ |
| description | Mannan, a highly abundant and cost-effective natural resource, holds great potential for the generation of high-value compounds such as bioactive polysaccharides and biofuels. In this study, we successfully enhanced the expression of constructed GH5 β-mannanase (TaMan5) from Trichoderma asperellum ND-1 by employing propeptide in Pichia pastoris. By replacing the α-factor with propeptide (MGNRALNSMKFFKSQALALLAATSAVA), TaMan5 activity was significantly increased from 67.5 to 91.7 U/mL. It retained higher activity in the presence of 20% ethanol and 15% NaCl. When incubated with a high concentration of mannotriose or mannotetraose, the transglycosylation action of TaMan5 can be detected, yielding the corresponding production of mannotetraose or mannooligosaccharides. Moreover, the unique mechanism whereby TaMan5 catalyzes the degradation of mannan into mannobiose involves the transglycosylation of mannose to mannotriose or mannotetraose as a substrate to produce a mannotetraose or mannopentose intermediate, respectively. Additionally, the production of soluble sugars from lignocellulose is a crucial step in bioethanol development, and it is noteworthy that TaMan5 could synergistically yield fermentable sugars from corn stover and bagasse. These findings offered valuable insights and strategies for enhancing β-mannanase expression and efficient conversion of lignocellulosic biomass, providing cost-effective and sustainable approaches for high-value biomolecule and biofuel production. |
| first_indexed | 2024-03-12T02:00:30Z |
| format | Article |
| id | doaj.art-af64637faf3e4e96843b400195e5b79d |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-03-12T02:00:30Z |
| publishDate | 2023-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-af64637faf3e4e96843b400195e5b79d2023-09-07T19:25:31ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852023-09-011110.3389/fbioe.2023.12447721244772Improved production of recombinant β-mannanase (TaMan5) in Pichia pastoris and its synergistic degradation of lignocellulosic biomassFengzhen Zheng0Abdul Basit1Zhiyue Zhang2Huan Zhuang3Jun Chen4Jianfen Zhang5College of Biological and Environmental Engineering, Zhejiang Shuren University, Hangzhou, ChinaDepartment of Microbiology, University of Jhang, Jhang, PakistanCollege of Biological and Environmental Engineering, Zhejiang Shuren University, Hangzhou, ChinaDepartment of ENT and Head and Neck Surgery, The Children’s Hospital Zhejiang University School of Medicine, Hangzhou, Zhejiang, ChinaInterdisciplinary Research Academy, Zhejiang Shuren University, Hangzhou, ChinaCollege of Biological and Environmental Engineering, Zhejiang Shuren University, Hangzhou, ChinaMannan, a highly abundant and cost-effective natural resource, holds great potential for the generation of high-value compounds such as bioactive polysaccharides and biofuels. In this study, we successfully enhanced the expression of constructed GH5 β-mannanase (TaMan5) from Trichoderma asperellum ND-1 by employing propeptide in Pichia pastoris. By replacing the α-factor with propeptide (MGNRALNSMKFFKSQALALLAATSAVA), TaMan5 activity was significantly increased from 67.5 to 91.7 U/mL. It retained higher activity in the presence of 20% ethanol and 15% NaCl. When incubated with a high concentration of mannotriose or mannotetraose, the transglycosylation action of TaMan5 can be detected, yielding the corresponding production of mannotetraose or mannooligosaccharides. Moreover, the unique mechanism whereby TaMan5 catalyzes the degradation of mannan into mannobiose involves the transglycosylation of mannose to mannotriose or mannotetraose as a substrate to produce a mannotetraose or mannopentose intermediate, respectively. Additionally, the production of soluble sugars from lignocellulose is a crucial step in bioethanol development, and it is noteworthy that TaMan5 could synergistically yield fermentable sugars from corn stover and bagasse. These findings offered valuable insights and strategies for enhancing β-mannanase expression and efficient conversion of lignocellulosic biomass, providing cost-effective and sustainable approaches for high-value biomolecule and biofuel production.https://www.frontiersin.org/articles/10.3389/fbioe.2023.1244772/fullTrichoderma asperellumβ-mannanaseimproved productiontransglycosylationcatalysis mechanismsynergism |
| spellingShingle | Fengzhen Zheng Abdul Basit Zhiyue Zhang Huan Zhuang Jun Chen Jianfen Zhang Improved production of recombinant β-mannanase (TaMan5) in Pichia pastoris and its synergistic degradation of lignocellulosic biomass Frontiers in Bioengineering and Biotechnology Trichoderma asperellum β-mannanase improved production transglycosylation catalysis mechanism synergism |
| title | Improved production of recombinant β-mannanase (TaMan5) in Pichia pastoris and its synergistic degradation of lignocellulosic biomass |
| title_full | Improved production of recombinant β-mannanase (TaMan5) in Pichia pastoris and its synergistic degradation of lignocellulosic biomass |
| title_fullStr | Improved production of recombinant β-mannanase (TaMan5) in Pichia pastoris and its synergistic degradation of lignocellulosic biomass |
| title_full_unstemmed | Improved production of recombinant β-mannanase (TaMan5) in Pichia pastoris and its synergistic degradation of lignocellulosic biomass |
| title_short | Improved production of recombinant β-mannanase (TaMan5) in Pichia pastoris and its synergistic degradation of lignocellulosic biomass |
| title_sort | improved production of recombinant β mannanase taman5 in pichia pastoris and its synergistic degradation of lignocellulosic biomass |
| topic | Trichoderma asperellum β-mannanase improved production transglycosylation catalysis mechanism synergism |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2023.1244772/full |
| work_keys_str_mv | AT fengzhenzheng improvedproductionofrecombinantbmannanasetaman5inpichiapastorisanditssynergisticdegradationoflignocellulosicbiomass AT abdulbasit improvedproductionofrecombinantbmannanasetaman5inpichiapastorisanditssynergisticdegradationoflignocellulosicbiomass AT zhiyuezhang improvedproductionofrecombinantbmannanasetaman5inpichiapastorisanditssynergisticdegradationoflignocellulosicbiomass AT huanzhuang improvedproductionofrecombinantbmannanasetaman5inpichiapastorisanditssynergisticdegradationoflignocellulosicbiomass AT junchen improvedproductionofrecombinantbmannanasetaman5inpichiapastorisanditssynergisticdegradationoflignocellulosicbiomass AT jianfenzhang improvedproductionofrecombinantbmannanasetaman5inpichiapastorisanditssynergisticdegradationoflignocellulosicbiomass |