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Published: 2025-09-18
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DOI: https://doi.org/10.52163/yhc.v66iCD14.3150
Issue
Vol. 66 No. CĐ14-NCKH Bệnh viện Giao thông vận tải
Section
Articles
How to Cite
Thu, P. D., Loan, P. T. B., Hiếu, B. M., Thảo, V. T., Thái, N. Q., & Minh, T. K. (2025). 43. INVESTIGATING CONDITION OF EXPRESSION CONDITIONS FOR RECOMBINANT HUMAN MONOAMINE OXIDASE B IN PICHIA PASTORIS. Vietnam Journal of Community Medicine, 66(CĐ14-NCKH Bệnh viện Giao thông vận tải). https://doi.org/10.52163/yhc.v66iCD14.3150

43. INVESTIGATING CONDITION OF EXPRESSION CONDITIONS FOR RECOMBINANT HUMAN MONOAMINE OXIDASE B IN PICHIA PASTORIS

Pham Diem Thu1, Phung Thi Bich Loan1, Bui Minh Hieu1, Vu Thanh Thao1, Nguyen Quoc Thai1, Thai Khac Minh2,3
1 School of Pharmacy, University of Medicine and Pharmacy, Ho Chi Minh City
2 University of Health Sciences - Ho Chi Minh City National University
3 Center for Research and Development of Health Care Products, VNU-HCM - Ho Chi Minh City National University

Main Article Content

Abstract

Objectives: To investigate and determine the optimal conditions for the expression of active recombinant human monoamine oxidase B (MAO-B) in Pichia pastoris KM71.


Methods: The P. pastoris KM71 strain carrying the gene encoding human MAO-B was used. Cultivation and induction conditions were investigated, including biomass-generating media (BMG, BMGY), induction media (BMM, BMMY), methanol concentrations (0.25%, 0.5%, 1%), and induction times (24 - 144 hours). Cell disruption conditions by mechanical bead beating were also optimized. MAO-B was purified by ion-exchange chromatography (HiTrap Q FF), and its activity was determined spectrophotometrically using benzylamine as a substrate.


Results: The minimal medium BMG - BMM yielded higher enzyme activity than the complex medium BMGY - BMMY. A methanol concentration of 0.5% and an induction time of 24 hours resulted in the highest MAO-B activity. The optimal cell disruption condition was 6 cycles of 6-minute beating and 10-minute incubation in ice bath. MAO-B was successfully purified, eluting mainly at 30% and 40% HIGH-QB concentrations, and the purified enzyme exhibited an activity of approximately 2977 U/L.


Conclusion: The study successfully expressed active recombinant human MAO-B in P. pastoris KM71 and identified optimal conditions for the expression, cell disruption, and initial purification processes, providing a foundation for further applications.

Article Details

Keywords

Human monoamine oxidase B, Pichia pastoris, recombinant

References

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