Open AccessEVALUATION OF RECOMBINANT GLUCOAMYLASE EXPRESSION BY A NATIVE AND α-MATING FACTOR SECRETION SIGNAL IN PICHIA PASTORIS
Author(s) -
Kazi Muhammad Rezaul Karim,
Tasmia Tasnim
Publication year - 2022
Publication title -
journal of microbiology, biotechnology and food sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.186
H-Index - 7
ISSN - 1338-5178
DOI - 10.55251/jmbfs.3428
Subject(s) - pichia pastoris , recombinant dna , signal peptide , pichia , yeast , expression vector , biochemistry , plasmid , biology , heterologous , secretion , aspergillus niger , heterologous expression , microbiology and biotechnology , chemistry , gene
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. 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. The constructed recombinant plasmids pPICZB_GA2 (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.