Abstract
To enhance the expression level of recombinant feruloyl esterase in Pichia pastoris, the Aspergillus niger feruloyl esterase (AnFaeA) gene was optimized based on the codon usage bias of P. pastoris and synthesized. The synthetic AnFaeA-syn was inserted into the pPICZαA and then integrated into P. pastoris X33. SDS-PAGE analysis of recombinant feruloyl esterase displayed a single protein band with an apparent molecular mass of 40.0 kDa. Western blot analysis confirmed that the expressed protein was the target protein. Furthermore, we showed that codon optimization increased enzyme yield for about 6.4-fold (35.12 U/mL vs 5.51 U/mL) in shake flask cultivation, compared to the non-codon-optimized control. Taken together, this study showed that the yield of feruloyl esterase could be increased by codon optimization.
Acknowledgements
This work was supported by the National Natural Science Foundation of China (No. 31201811) and the Specific Research Supporting Program for Discipline Construction at Sichuan Agricultural University.
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- Abbreviations
- AnFaeA
Aspergillus niger feruloyl esterase
- AOX1
alcohol oxidase
- FA
ferulic acid.
©2016 Institute of Molecular Biology, Slovak Academy of Sciences
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Articles in the same Issue
- Cellular and Molecular Biology
- Molecular detection of Mycobacterium tuberculosis complex in the 8th century skeletal remains from the territory of Slovakia
- Cellular and Molecular Biology
- First report of microorganisms of Caucasus glaciers (Georgia)
- Cellular and Molecular Biology
- Codon optimization of Aspergillus niger feruloyl esterase and its expression in Pichia pastoris
- Botany
- Response of lichens Cladonia arbuscula subsp. mitis and Cladonia furcata to nitrogen excess
- Botany
- No confirmation for previously suggested presence of diploid cytotypes of Sesleria (Poaceae) on the Balkan Peninsula
- Botany
- RCD1 homologues and their constituent WWE domain in plants: analysis of conservation through phylogeny methods
- Botany
- Nucleoli migration coupled with cytomixis
- Cellular and Molecular Biology
- Evaluation of appropriate reference gene for normalization of microRNA expression by real-time PCR in Lablab purpureus under abiotic stress conditions
- Zoology
- The fractal nature of the latitudinal biodiversity gradient
- Zoology
- A new species of Neoribates (Neoribates) (Acari: Oribatida: Parakalummidae) with key to the Neotropical species of the subgenus
- Zoology
- Diversity patterns of aquatic specialists and generalists: contrasts among two spring-fen mesohabitats and nearby streams
- Zoology
- Heteroptera (Insecta: Hemiptera) of the peat bogs of Belarusian Lakeland
- Cellular and Molecular Biology
- Cloning of monoacylglycerol o-acyltransferase 2 cDNA from a silkworm, Bombyx mori
- Zoology
- Biological aspect of the surface structure of the tongue in the adult red kangaroo (Macropus rufus) — light and scanning electron microscopy
- Zoology
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- Zoology
- Considerations on the vulnerability of the Eurasian water shrew Neomys fodiens to the presence of introduced brown trout Salmo trutta