TAILIEUCHUNG - Biochemical and in silico evaluation of recombinant E. coli aminopeptidase and in vitro processed human interferon α-2b

Escherichia coli is an extensively used host for the production of recombinant proteins, making its N-terminal methionine aminopeptidase (MAP) an attractive candidate for studies on posttranslational protein processing. The present study describes the recombinant production and properties of MAP from the DH5α strain of E. coli. | Turkish Journal of Biology Research Article Turk J Biol (2018) 42: 240-249 © TÜBİTAK doi: Biochemical and in silico evaluation of recombinant E. coli aminopeptidase and in vitro processed human interferon α-2b 1 2 2, Amina ARIF , Kaleemuddin MOHAMMED , Muhammad Shahid NADEEM * 1 Faculty of Life Sciences, University of Central Punjab, Lahore, Pakistan 2 Department of Biochemistry, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia Received: Accepted/Published Online: Final Version: Abstract: Escherichia coli is an extensively used host for the production of recombinant proteins, making its N-terminal methionine aminopeptidase (MAP) an attractive candidate for studies on posttranslational protein processing. The present study describes the recombinant production and properties of MAP from the DH5α strain of E. coli. The soluble and active enzyme was produced in E. coli BL21 (DE3) RIL - codon plus cells under a T7 promoter system and purified by anion-exchange chromatography. It exhibited a molecular weight of 29, Da by MALDI-TOF analysis. The purified enzyme showed specific activity of U/mg with methionylp-nitroanilide and U/mg with synthetic tetrapeptide substrate ‘MGMM’ in a discontinuous HPLC-based assay. In vitro studies showed the processing of up to 36% of Met-INFα-2b in 40 min. In silico studies revealed that the ES-complex formation between the enzyme and interferon has a ΔG – kJ/mol. Molecular docking results showed that the processed INFα-2b has greater binding affinity with IFNAR2 receptor as indicated by ΔG – kJ/mol, significantly lower than that of methionine containing INFα-2b (ΔG – kJ/mol). These findings emphasize the functional superiority or better efficacy of N-terminal methionine processed recombinant interferon. Key words: Methionine aminopeptidase, E. coli, human interferon, in vitro, in .

TỪ KHÓA LIÊN QUAN
Đã phát hiện trình chặn quảng cáo AdBlock
Trang web này phụ thuộc vào doanh thu từ số lần hiển thị quảng cáo để tồn tại. Vui lòng tắt trình chặn quảng cáo của bạn hoặc tạm dừng tính năng chặn quảng cáo cho trang web này.