Recombinant Expression and Bioactivity Comparison of Four Typical Fungal Immunomodulatory Proteins from Three Main Ganoderma Species
Abstract Background More than a dozen of fungal immunomodulatory proteins (FIPs) have been identified to date, most of which are from Ganoderma species. However, little is known about the similarities and differences between different Ganoderma FIPs’ bioactivities. In the current study, two FIP gene...
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doaj-59cd9f6c33184a92b046df9b7e0293bd2020-11-25T03:55:58ZengBMCBMC Biotechnology1472-67502018-12-0118111210.1186/s12896-018-0488-0Recombinant Expression and Bioactivity Comparison of Four Typical Fungal Immunomodulatory Proteins from Three Main Ganoderma SpeciesZheng-Wei Qu0Si-Ya Zhou1Shi-Xin Guan2Rui Gao3Zuo-Wen Duan4Xin Zhang5Wei-Yan Sun6Wen-Li Fan7Shui-Sen Chen8Li-Jing Chen9Jing-Wei Lin10Yan-Ye Ruan11Liaoning Province Key Laboratory of Agricultural Technology, College of Bioscience and Biotechnology, Shenyang Agricultural UniversityLiaoning Province Key Laboratory of Agricultural Technology, College of Bioscience and Biotechnology, Shenyang Agricultural UniversityCollege of Horticulture, Shenyang Agricultural UniversityLiaoning Province Key Laboratory of Agricultural Technology, College of Bioscience and Biotechnology, Shenyang Agricultural UniversityLiaoning Province Key Laboratory of Agricultural Technology, College of Bioscience and Biotechnology, Shenyang Agricultural UniversityLiaoning Province Key Laboratory of Agricultural Technology, College of Bioscience and Biotechnology, Shenyang Agricultural UniversityLiaoning Province Key Laboratory of Agricultural Technology, College of Bioscience and Biotechnology, Shenyang Agricultural UniversityCollege of Horticulture, Shenyang Agricultural UniversityLiaoning Province Key Laboratory of Agricultural Technology, College of Bioscience and Biotechnology, Shenyang Agricultural UniversityLiaoning Province Key Laboratory of Agricultural Technology, College of Bioscience and Biotechnology, Shenyang Agricultural UniversityLiaoning Province Key Laboratory of Agricultural Technology, College of Bioscience and Biotechnology, Shenyang Agricultural UniversityLiaoning Province Key Laboratory of Agricultural Technology, College of Bioscience and Biotechnology, Shenyang Agricultural UniversityAbstract Background More than a dozen of fungal immunomodulatory proteins (FIPs) have been identified to date, most of which are from Ganoderma species. However, little is known about the similarities and differences between different Ganoderma FIPs’ bioactivities. In the current study, two FIP genes termed FIP-gap1 and FIP-gap2 from G. applanatum, along with LZ-8 and FIP-gsi, another two representative Ganoderma FIP genes from G. lucidum and G. sinense were functionally expressed in Pichia. Subsequently, bioactivities of four recombinant Ganoderma FIPs were demonstrated and compared. Results All the four Ganoderma FIP genes could be effectively expressed in P. pastoris GS115 at expression levels ranging from 197.5 to 264.3 mg L− 1 and simply purified by one step chromatography using HisTrap™ FF prepack columns. Amino acid sequence analysis showed that they all possessed the FIP conserved fragments. The homologies of different Ganoderma FIPs were from 72.6 to 86.4%. In vitro haemagglutination exhibited that FIP-gap1, FIP-gsi and LZ-8 could agglutinate human, sheep and mouse red blood cells but FIP-gap2 agglutinated none. Besides, the immunomodulation activities of these Ganoderma FIPs were as: rFIP-gap2 > rFIP-gap1 > rLZ-8 and rFIP-gsi in terms of proliferation stimulation and cytokine induction on murine splenocytes. Additionally, the cytotoxic activity of different FIPs was: rFIP-gap1 > rLZ-8 > rFIP-gsi > rFIP-gap2, examined by their inhibition of three human carcinomas A549, Hela and MCF-7. Conclusions Taken together, four typical Ganoderma FIP genes could be functionally expressed in P. pastoris, which might supply as feasible efficient resources for further study and application. Both similarities and differences were indeed observed between Ganoderma FIPs in their amino acid sequences and bioactivities. Comprehensively, rFIP-gaps from G. applanatum proved to be more effective in immunomodulation and cytotoxic assays in vitro than rLZ-8 (G. lucidum) and rFIP-gsi (G. sinense).http://link.springer.com/article/10.1186/s12896-018-0488-0GanodermaFungal immunomodulatory proteinRecombinant expressionCytotoxicityYeast |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Zheng-Wei Qu Si-Ya Zhou Shi-Xin Guan Rui Gao Zuo-Wen Duan Xin Zhang Wei-Yan Sun Wen-Li Fan Shui-Sen Chen Li-Jing Chen Jing-Wei Lin Yan-Ye Ruan |
spellingShingle |
Zheng-Wei Qu Si-Ya Zhou Shi-Xin Guan Rui Gao Zuo-Wen Duan Xin Zhang Wei-Yan Sun Wen-Li Fan Shui-Sen Chen Li-Jing Chen Jing-Wei Lin Yan-Ye Ruan Recombinant Expression and Bioactivity Comparison of Four Typical Fungal Immunomodulatory Proteins from Three Main Ganoderma Species BMC Biotechnology Ganoderma Fungal immunomodulatory protein Recombinant expression Cytotoxicity Yeast |
author_facet |
Zheng-Wei Qu Si-Ya Zhou Shi-Xin Guan Rui Gao Zuo-Wen Duan Xin Zhang Wei-Yan Sun Wen-Li Fan Shui-Sen Chen Li-Jing Chen Jing-Wei Lin Yan-Ye Ruan |
author_sort |
Zheng-Wei Qu |
title |
Recombinant Expression and Bioactivity Comparison of Four Typical Fungal Immunomodulatory Proteins from Three Main Ganoderma Species |
title_short |
Recombinant Expression and Bioactivity Comparison of Four Typical Fungal Immunomodulatory Proteins from Three Main Ganoderma Species |
title_full |
Recombinant Expression and Bioactivity Comparison of Four Typical Fungal Immunomodulatory Proteins from Three Main Ganoderma Species |
title_fullStr |
Recombinant Expression and Bioactivity Comparison of Four Typical Fungal Immunomodulatory Proteins from Three Main Ganoderma Species |
title_full_unstemmed |
Recombinant Expression and Bioactivity Comparison of Four Typical Fungal Immunomodulatory Proteins from Three Main Ganoderma Species |
title_sort |
recombinant expression and bioactivity comparison of four typical fungal immunomodulatory proteins from three main ganoderma species |
publisher |
BMC |
series |
BMC Biotechnology |
issn |
1472-6750 |
publishDate |
2018-12-01 |
description |
Abstract Background More than a dozen of fungal immunomodulatory proteins (FIPs) have been identified to date, most of which are from Ganoderma species. However, little is known about the similarities and differences between different Ganoderma FIPs’ bioactivities. In the current study, two FIP genes termed FIP-gap1 and FIP-gap2 from G. applanatum, along with LZ-8 and FIP-gsi, another two representative Ganoderma FIP genes from G. lucidum and G. sinense were functionally expressed in Pichia. Subsequently, bioactivities of four recombinant Ganoderma FIPs were demonstrated and compared. Results All the four Ganoderma FIP genes could be effectively expressed in P. pastoris GS115 at expression levels ranging from 197.5 to 264.3 mg L− 1 and simply purified by one step chromatography using HisTrap™ FF prepack columns. Amino acid sequence analysis showed that they all possessed the FIP conserved fragments. The homologies of different Ganoderma FIPs were from 72.6 to 86.4%. In vitro haemagglutination exhibited that FIP-gap1, FIP-gsi and LZ-8 could agglutinate human, sheep and mouse red blood cells but FIP-gap2 agglutinated none. Besides, the immunomodulation activities of these Ganoderma FIPs were as: rFIP-gap2 > rFIP-gap1 > rLZ-8 and rFIP-gsi in terms of proliferation stimulation and cytokine induction on murine splenocytes. Additionally, the cytotoxic activity of different FIPs was: rFIP-gap1 > rLZ-8 > rFIP-gsi > rFIP-gap2, examined by their inhibition of three human carcinomas A549, Hela and MCF-7. Conclusions Taken together, four typical Ganoderma FIP genes could be functionally expressed in P. pastoris, which might supply as feasible efficient resources for further study and application. Both similarities and differences were indeed observed between Ganoderma FIPs in their amino acid sequences and bioactivities. Comprehensively, rFIP-gaps from G. applanatum proved to be more effective in immunomodulation and cytotoxic assays in vitro than rLZ-8 (G. lucidum) and rFIP-gsi (G. sinense). |
topic |
Ganoderma Fungal immunomodulatory protein Recombinant expression Cytotoxicity Yeast |
url |
http://link.springer.com/article/10.1186/s12896-018-0488-0 |
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