Combined strategies for improving the heterologous expression of a novel xylanase from Fusarium oxysporum Fo47 in Pichia pastoris
Xylanase, an enzyme capable of hydrolyzing non-starch polysaccharides found in grain structures like wheat, has been found to improve the organizational structure of dough and thus increase its volume. In our past work, one promising xylanase FXYL derived from Fusarium oxysporum Fo47 and first expre...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
KeAi Communications Co., Ltd.
2024-09-01
|
| Series: | Synthetic and Systems Biotechnology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405805X24000462 |
| _version_ | 1797219331634888704 |
|---|---|
| author | Chun Liu Yaping Zhang Chunting Ye Fengguang Zhao Yian Chen Shuangyan Han |
| author_facet | Chun Liu Yaping Zhang Chunting Ye Fengguang Zhao Yian Chen Shuangyan Han |
| author_sort | Chun Liu |
| collection | DOAJ |
| description | Xylanase, an enzyme capable of hydrolyzing non-starch polysaccharides found in grain structures like wheat, has been found to improve the organizational structure of dough and thus increase its volume. In our past work, one promising xylanase FXYL derived from Fusarium oxysporum Fo47 and first expressed 779.64 U/mL activity in P. pastoris. It has shown significant potential in improving the quality of whole wheat bread, making it become a candidate for development as a new flour improver. After optimization of expression elements and gene dose, the xylanase activity of FXYL strain carrying three-copies reached 4240.92 U/mL in P. pastoris. In addition, 12 factors associated with the three stages of protein expression pathway were co-expressed individually in order in three-copies strain, and the translation factor Pab1 co-expression increased FXYL activity to 8893.53 U/mL. Nevertheless, combining the most effective or synergistic factors from three stages did not exhibit better results than co-expressing them alone. To further evaluate the industrial potential, the xylanase activity and protein concentration reached 81184.51 U/mL and 11.8 g/L in a 5 L fed-batch fermenter. These engineering strategies improved the expression of xylanase FXYL by more than 104-fold, providing valuable insights for the cost-effective industrial application of FXYL in the baking field. |
| first_indexed | 2024-04-24T12:31:57Z |
| format | Article |
| id | doaj.art-976a5358e2cf499f944252bb8843ee2f |
| institution | Directory Open Access Journal |
| issn | 2405-805X |
| language | English |
| last_indexed | 2024-04-24T12:31:57Z |
| publishDate | 2024-09-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Synthetic and Systems Biotechnology |
| spelling | doaj.art-976a5358e2cf499f944252bb8843ee2f2024-04-08T04:08:32ZengKeAi Communications Co., Ltd.Synthetic and Systems Biotechnology2405-805X2024-09-0193426435Combined strategies for improving the heterologous expression of a novel xylanase from Fusarium oxysporum Fo47 in Pichia pastorisChun Liu0Yaping Zhang1Chunting Ye2Fengguang Zhao3Yian Chen4Shuangyan Han5School of Biology and Biological Engineering, South China University of Technology, Guangzhou, 510640, ChinaSchool of Biology and Biological Engineering, South China University of Technology, Guangzhou, 510640, ChinaSchool of Biology and Biological Engineering, South China University of Technology, Guangzhou, 510640, ChinaSchool of Light Industry and Engineering, South China University of Technology, Guangzhou, 510640, ChinaSchool of Light Industry and Engineering, South China University of Technology, Guangzhou, 510640, ChinaSchool of Biology and Biological Engineering, South China University of Technology, Guangzhou, 510640, China; Corresponding author.Xylanase, an enzyme capable of hydrolyzing non-starch polysaccharides found in grain structures like wheat, has been found to improve the organizational structure of dough and thus increase its volume. In our past work, one promising xylanase FXYL derived from Fusarium oxysporum Fo47 and first expressed 779.64 U/mL activity in P. pastoris. It has shown significant potential in improving the quality of whole wheat bread, making it become a candidate for development as a new flour improver. After optimization of expression elements and gene dose, the xylanase activity of FXYL strain carrying three-copies reached 4240.92 U/mL in P. pastoris. In addition, 12 factors associated with the three stages of protein expression pathway were co-expressed individually in order in three-copies strain, and the translation factor Pab1 co-expression increased FXYL activity to 8893.53 U/mL. Nevertheless, combining the most effective or synergistic factors from three stages did not exhibit better results than co-expressing them alone. To further evaluate the industrial potential, the xylanase activity and protein concentration reached 81184.51 U/mL and 11.8 g/L in a 5 L fed-batch fermenter. These engineering strategies improved the expression of xylanase FXYL by more than 104-fold, providing valuable insights for the cost-effective industrial application of FXYL in the baking field.http://www.sciencedirect.com/science/article/pii/S2405805X24000462XylanasePichia pastorisHeterologous proteinSecretion expression |
| spellingShingle | Chun Liu Yaping Zhang Chunting Ye Fengguang Zhao Yian Chen Shuangyan Han Combined strategies for improving the heterologous expression of a novel xylanase from Fusarium oxysporum Fo47 in Pichia pastoris Synthetic and Systems Biotechnology Xylanase Pichia pastoris Heterologous protein Secretion expression |
| title | Combined strategies for improving the heterologous expression of a novel xylanase from Fusarium oxysporum Fo47 in Pichia pastoris |
| title_full | Combined strategies for improving the heterologous expression of a novel xylanase from Fusarium oxysporum Fo47 in Pichia pastoris |
| title_fullStr | Combined strategies for improving the heterologous expression of a novel xylanase from Fusarium oxysporum Fo47 in Pichia pastoris |
| title_full_unstemmed | Combined strategies for improving the heterologous expression of a novel xylanase from Fusarium oxysporum Fo47 in Pichia pastoris |
| title_short | Combined strategies for improving the heterologous expression of a novel xylanase from Fusarium oxysporum Fo47 in Pichia pastoris |
| title_sort | combined strategies for improving the heterologous expression of a novel xylanase from fusarium oxysporum fo47 in pichia pastoris |
| topic | Xylanase Pichia pastoris Heterologous protein Secretion expression |
| url | http://www.sciencedirect.com/science/article/pii/S2405805X24000462 |
| work_keys_str_mv | AT chunliu combinedstrategiesforimprovingtheheterologousexpressionofanovelxylanasefromfusariumoxysporumfo47inpichiapastoris AT yapingzhang combinedstrategiesforimprovingtheheterologousexpressionofanovelxylanasefromfusariumoxysporumfo47inpichiapastoris AT chuntingye combinedstrategiesforimprovingtheheterologousexpressionofanovelxylanasefromfusariumoxysporumfo47inpichiapastoris AT fengguangzhao combinedstrategiesforimprovingtheheterologousexpressionofanovelxylanasefromfusariumoxysporumfo47inpichiapastoris AT yianchen combinedstrategiesforimprovingtheheterologousexpressionofanovelxylanasefromfusariumoxysporumfo47inpichiapastoris AT shuangyanhan combinedstrategiesforimprovingtheheterologousexpressionofanovelxylanasefromfusariumoxysporumfo47inpichiapastoris |