[The analysis of heterogeneity of HWTX-I expressed in Pichia pastoris].
Sheng Wu Gong Cheng Xue Bao
; 22(2): 215-9, 2006 Mar.
Article
in Zh
| MEDLINE
| ID: mdl-16607946
ABSTRACT
To seek the reason of heterogeneity of recombinant HWTX-I (rHWTX-I) expressed in Pichia pastoris. We expressed HWTX-I gene of interest in Pichia pastoris GS115/HWTX-I. The heterogenous product expressed was separated, purified and identified by using Ion exchange HPLC, reverse HPLC, Tricine SDS-PAGE and MALDI-TOF Mass Spectrometry and then sequenced in both N-terminus and C-terminus. These results show that the heterogeneity of rHWTX-I results from the incomplete processing of signal peptide of N-terminus and the internal degradation of C-terminus. Biological activity assay shows that the activity of the heterogenous rHWTX-I only showed 30% activity compared with the native HWTX-I. The Solutions to how to avoid the heterogeneity are also discussed.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Pichia
/
Spider Venoms
/
Reptilian Proteins
/
Neurotoxins
Type of study:
Prognostic_studies
Limits:
Animals
Language:
Zh
Journal:
Sheng Wu Gong Cheng Xue Bao
Journal subject:
BIOTECNOLOGIA
Year:
2006
Document type:
Article
Affiliation country:
China