Molecular expression of a recombinant thermostable bacterial amylase from geobacillus stearothermophilus SR74 using methanol-free meyerozyma guilliermondii strain SO yeast system
α-Amylase, which was isolated from Geobacillus stearothermophilus SR74, has shown its potential to be used in industrial applications. However, its expression in the Pichia pastoris expression system with the alcohol oxidase 1 promoter (PAOX1) requires high methanol consumption and is time-consuming...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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College of Natural Resources, North Carolina State University
2020
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| Online Access: | http://psasir.upm.edu.my/id/eprint/86820/ http://psasir.upm.edu.my/id/eprint/86820/1/Molecular%20expression%20of%20a%20recombinant%20thermostable%20bacterial.pdf |
| _version_ | 1848860295683899392 |
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| author | Mohamad Nasir, Nurul Syazwani Chor, Thean Leow Oslan, Siti N.H. Salleh, Abu Bakar Oslan, Siti Nurbaya |
| author_facet | Mohamad Nasir, Nurul Syazwani Chor, Thean Leow Oslan, Siti N.H. Salleh, Abu Bakar Oslan, Siti Nurbaya |
| author_sort | Mohamad Nasir, Nurul Syazwani |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | α-Amylase, which was isolated from Geobacillus stearothermophilus SR74, has shown its potential to be used in industrial applications. However, its expression in the Pichia pastoris expression system with the alcohol oxidase 1 promoter (PAOX1) requires high methanol consumption and is time-consuming. This study aimed to express SR74 α-amylase in an alternative yeast system, using Meyerozyma guilliermondii strain SO, which was isolated from a spoiled orange (SO) under the regulation of a formaldehyde dehydrogenase promoter (PFLD). Qualitative screening showed that strain SO possessed a native amylase grown on YPD-starch plate at 30 °C. The recombinant SR74 α-amylase was further quantified and validated using the Western blot test. It was confirmed that SR74 α-amylase was expressed by strain SO extracellularly with a size of 59 kDa. Optimization in a shake flask showed that the recombinant SR74 α-amylase, which was regulated by PFLD, was successfully produced (26 U/mL) without any external inducer in the YPT medium after 24 h of cultivation. In conclusion, strain SO was able to produce SR74 amylase without methanol in one-fifth the fermentation time of P. pastoris. Further optimization of the expression may be done to improve the yield, as this methanol-free host is still underexplored. |
| first_indexed | 2025-11-15T12:42:58Z |
| format | Article |
| id | upm-86820 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T12:42:58Z |
| publishDate | 2020 |
| publisher | College of Natural Resources, North Carolina State University |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-868202021-11-17T10:43:56Z http://psasir.upm.edu.my/id/eprint/86820/ Molecular expression of a recombinant thermostable bacterial amylase from geobacillus stearothermophilus SR74 using methanol-free meyerozyma guilliermondii strain SO yeast system Mohamad Nasir, Nurul Syazwani Chor, Thean Leow Oslan, Siti N.H. Salleh, Abu Bakar Oslan, Siti Nurbaya α-Amylase, which was isolated from Geobacillus stearothermophilus SR74, has shown its potential to be used in industrial applications. However, its expression in the Pichia pastoris expression system with the alcohol oxidase 1 promoter (PAOX1) requires high methanol consumption and is time-consuming. This study aimed to express SR74 α-amylase in an alternative yeast system, using Meyerozyma guilliermondii strain SO, which was isolated from a spoiled orange (SO) under the regulation of a formaldehyde dehydrogenase promoter (PFLD). Qualitative screening showed that strain SO possessed a native amylase grown on YPD-starch plate at 30 °C. The recombinant SR74 α-amylase was further quantified and validated using the Western blot test. It was confirmed that SR74 α-amylase was expressed by strain SO extracellularly with a size of 59 kDa. Optimization in a shake flask showed that the recombinant SR74 α-amylase, which was regulated by PFLD, was successfully produced (26 U/mL) without any external inducer in the YPT medium after 24 h of cultivation. In conclusion, strain SO was able to produce SR74 amylase without methanol in one-fifth the fermentation time of P. pastoris. Further optimization of the expression may be done to improve the yield, as this methanol-free host is still underexplored. College of Natural Resources, North Carolina State University 2020 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/86820/1/Molecular%20expression%20of%20a%20recombinant%20thermostable%20bacterial.pdf Mohamad Nasir, Nurul Syazwani and Chor, Thean Leow and Oslan, Siti N.H. and Salleh, Abu Bakar and Oslan, Siti Nurbaya (2020) Molecular expression of a recombinant thermostable bacterial amylase from geobacillus stearothermophilus SR74 using methanol-free meyerozyma guilliermondii strain SO yeast system. BioResources, 15 (2). 3161 - 3172. ISSN 1930-2126 https://ojs.cnr.ncsu.edu/index.php/BioRes/article/view/BioRes_15_2_3161_Mohamad_Nasir_Molecular_Expression_Recombinant_Thermostable |
| spellingShingle | Mohamad Nasir, Nurul Syazwani Chor, Thean Leow Oslan, Siti N.H. Salleh, Abu Bakar Oslan, Siti Nurbaya Molecular expression of a recombinant thermostable bacterial amylase from geobacillus stearothermophilus SR74 using methanol-free meyerozyma guilliermondii strain SO yeast system |
| title | Molecular expression of a recombinant thermostable bacterial amylase from geobacillus stearothermophilus SR74 using methanol-free meyerozyma guilliermondii strain SO yeast system |
| title_full | Molecular expression of a recombinant thermostable bacterial amylase from geobacillus stearothermophilus SR74 using methanol-free meyerozyma guilliermondii strain SO yeast system |
| title_fullStr | Molecular expression of a recombinant thermostable bacterial amylase from geobacillus stearothermophilus SR74 using methanol-free meyerozyma guilliermondii strain SO yeast system |
| title_full_unstemmed | Molecular expression of a recombinant thermostable bacterial amylase from geobacillus stearothermophilus SR74 using methanol-free meyerozyma guilliermondii strain SO yeast system |
| title_short | Molecular expression of a recombinant thermostable bacterial amylase from geobacillus stearothermophilus SR74 using methanol-free meyerozyma guilliermondii strain SO yeast system |
| title_sort | molecular expression of a recombinant thermostable bacterial amylase from geobacillus stearothermophilus sr74 using methanol-free meyerozyma guilliermondii strain so yeast system |
| url | http://psasir.upm.edu.my/id/eprint/86820/ http://psasir.upm.edu.my/id/eprint/86820/ http://psasir.upm.edu.my/id/eprint/86820/1/Molecular%20expression%20of%20a%20recombinant%20thermostable%20bacterial.pdf |