pPIC9K-anPhyA

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	The resultant recombinant P. pastoris was inoculated in a 1-L Erlenmeyer flask with 200 mL of standard medium (1.34% [w/v] yeast nitrogen base, 2% [w/v] peptone, 1% [w/v] yeast extract, 0.4 mg/L biotin, and 100 mM potassium phosphate [pH 6.0]) containing 1% (w/v) glycerol (Buffered Glycerol-complex Medium). The inocula were placed in an incubator shaker at 29°C/250 rpm for 36–38 h until a wet cell weight of 30 g/L was achieved.

Protocol tips

The resultant recombinant P. pastoris was inoculated in a 1-L Erlenmeyer flask with 200 mL of standard medium (1.34% [w/v] yeast nitrogen base, 2% [w/v] peptone, 1% [w/v] yeast extract, 0.4 mg/L biotin, and 100 mM potassium phosphate [pH 6.0]) containing 1% (w/v) glycerol (Buffered Glycerol-complex Medium). The inocula were placed in an incubator shaker at 29°C/250 rpm for 36–38 h until a wet cell weight of 30 g/L was achieved.

Publication protocol

"The Mut+ strain of Pichia pastoris (Komagataella phaffii) GS115 (His4) was used to express Aspergillus niger PhyA phytase under the control of the inducible AOX1 promoter. The alpha-mating factor secretion signal of S. cerevisiae was also incorporated into the plasmid construct (Han et al., 2018). P. pastoris was grown in liquid and solid media during fermentation in accordance with the procedure used by Rahimi et al. (2019) with slight modifications.

The resultant recombinant P. pastoris was inoculated in a 1-L Erlenmeyer flask with 200 mL of standard medium (1.34% [w/v] yeast nitrogen base, 2% [w/v] peptone, 1% [w/v] yeast extract, 0.4 mg/L biotin, and 100 mM potassium phosphate [pH 6.0]) containing 1% (w/v) glycerol (Buffered Glycerol-complex Medium). The inocula were placed in an incubator shaker at 29°C/250 rpm for 36–38 h until a wet cell weight of 30 g/L was achieved."

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Protein Expression Eukaryotic cells - P. pastoris Aspergillus niger PhyA using pPIC9K-anPhyA from Zunxi Huang, Engineering Research Center of Sustainable Developm.

Manufacturer protocol

Download the product protocol from Zunxi Huang, Engineering Research Center of Sustainable Developm for pPIC9K-anPhyA below.

Videos

Check out videos that might be relevant for performing Protein Expression Eukaryotic cells - P. pastoris Aspergillus niger PhyA using pPIC9K-anPhyA from Zunxi Huang, Engineering Research Center of Sustainable Developm. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.