Cloning, functional expression and characterization of Aspergillus sulphureus β-mannanase in Pichia pastoris

Cloning and expression of a family 10 xylanase gene (Aoxyn10) from Aspergillus oryzae in Pichia pastoris

“…It was reported that the purities of recombinant A. usamii xylanase AuXyn11D and A. sulphureus β-mannanase expressed in P. pastoris GS115 and X-33 reached 90 and 97%, respectively (Chen et al, 2007;Zhang et al, 2012). In this work, the expressed reAoXyn10 accounted for more than 88% of the total protein in the cultured supernatant, which was analyzed by protein band-scanning.…”

Overexpression of an optimized Aspergillus sulphureus β-mannanase gene in Pichia pastoris

“…The full-length cDNA sequence of A. sulphureusβ-mannanase gene was cloned in our previous study (Chen et al 2007). Then according to the codon bias of P. pastoris (Zhao et al 2000), the wild type β-mannanase gene (manWT) was optimized without amino acid alteration.…”

“…• C which last for 6 d. P. pastoris X-33 harbored the manWT, described previously (Chen et al 2007), was used as a control. To detect the stability of the heterogeneous gene, the yeast was successively inoculated in yeast extract peptone dextrose medium for 20 passages.…”

“…In our lab, an A. sulphureus β-mannanase gene has been cloned and expressed in Pichia pastoris (Chen et al 2007). In an effort to further increase the expression of this β-mannanase, the main goal of the present study was to modify the gene sequence according to codon bias of P. pastoris and assay the expressed enzyme activity.…”

Advanced Strategies for Improving the Production of Industrial Enzymes in Heterologous Host Systems

“…With the development of genetic engineering, a lot of studies have been carried out to improve the production of industrial enzymes from the molecular level. Genes of some enzymes, such as glucoamylase, amylase, lipase, cellulase and phytase, etc., have been cloned and expressed efficiently in heterologous host systems, such as Saccharomyces cerevisiae; Aspergillus niger and Pichia pastoris [1][2][3][4][5][6][7][8][9][10][11]. In this paper, advanced strategies for improving the production of industrial enzymes were reviewed from the following aspects: the introduction of the strong promoter, increasing the copy number of genes, changing the signal peptide sequence and designing the codon bias ( Figure 1).…”