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Evaluation of recombinant glucoamylase expression by a native and α-mating factor secretion signal in pichia pastoris

The purpose of the study was to measure the secretion levels of recombinant glucoamylase from Pichia pastoris, by using the α-mating factor secretion signal peptide (α-MF) and the native signal peptide of glucoamylase from Aspergillus flavus NSH9. The Aspergillus flavus NSH9 gene (with and without native signal sequences), encoding a pH and thermostable glucoamylase with an SBD, was successfully cloned and expressed in Pichia pastoris to produce recombinant glucoamylases. | EVALUATION OF RECOMBINANT GLUCOAMYLASE EXPRESSION BY A NATIVE AND α-MATING FACTOR SECRETION SIGNAL IN PICHIA PASTORIS Kazi Muhammad Rezaul Karim1 and Tasmia Tasnim2 Address es Kazi Muhammad Rezaul Karim Ph.D. 1 Institute of Nutrition and Food Science University of Dhaka Dhaka-1000 Bangladesh. 2 Department of Nutrition and Food Engineering Faculty of Allied Health Science Daffodil International University Dhaka-1207 Bangladesh. Corresponding author rkarim98@gmail.com rezaul.infs@du.ac.bd https doi.org 10.15414 jmbfs.3428 ARTICLE INFO ABSTRACT Received 11. 7. 2021 Raw starch degrading enzyme specially glucoamylase with starch binding domain SBD has great values in the starch processing industry because it digests the starch particles below the gelatinization temperature by releasing glucose from the non-reducing ends sequentially. Revised 10. 11. 2021 The purpose of the study was to measure the secretion levels of recombinant glucoamylase from Pichia pastoris by using the α-mating Accepted 12. 11. 2021 factor secretion signal peptide α-MF and the native signal peptide of glucoamylase from Aspergillus flavus NSH9. The Aspergillus flavus Published xx.xx.201x NSH9 gene with and without native signal sequences encoding a pH and thermostable glucoamylase with an SBD was successfully cloned and expressed in Pichia pastoris to produce recombinant glucoamylases. The constructed recombinant plasmids pPICZB_GA2 Regular article having a native signal peptides and pPICZαC_GA2 having the α-MF were 5144 and 5356 bp in length respectively. Recombinant pichia having α-MF signal sequence plasmid pPICZαC_GA2 gave the highest level of secretions of recombinant glucoamylase after 6 days of incubation period with 0.5 methanol. In conclusion yeast expression vector signal peptide is more efficient for heterologous expression secretions of recombinant glucoamylase compared to its native signal sequences. Keywords Raw starch degrading glucoamylase signal peptide Pichia pastoris Aspergillus