Molecular expression of a recombinant thermostable bacterial amylase from Geobacillus stearothermophilus SR74 using methanol-free Meyerozyma guilliermondii strain SO yeast system

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...

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Bibliographic Details
Published in:Bioresources Vol. 15; no. 2; pp. 3161 - 3172
Main Authors: Nasir, Nurul S. M., Leow, Chor T., Oslan, Siti N. H., Salleh, Abu B., Oslan, Siti N.
Format: Journal Article
Language:English
Published: Raleigh North Carolina State University 01-05-2020
Subjects:
Alcohol oxidase
Amylases
Cloning
E coli
Enzymes
Fermentation
Formaldehyde dehydrogenase
Geobacillus stearothermophilus
Glycerol
Industrial applications
Iodine
Methanol
Meyerozyma guilliermondii
Optimization
Plasmids
Potassium
Proteins
Starch
Yeast
Yeasts
α-Amylase
United States > US
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Summary: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.
ISSN:1930-2126
1930-2126
DOI:10.15376/biores.15.2.3161-3172