تلخیص
The importance of chitinases over the years had attracted huge biotechnological attention
because its usage cut across wide range of field. It plays a significant role in the defensive mechanism
against fungal pathogens.
Methods: In this study, an endochitinase gene was isolated from Trichoderma harzianum, and characterized insilico by using various bioinformatics tools. Further, the gene was cloned in eukaryotic expression vector
(pPICZA) under the control of AOX1 promoter for recombinant expression in Pichia pastoris GS115 host strain.
Results: The chitinase cDNA was ~1000 bp long, while in-silico studies revealed an open reading frame of 888
bp encoding 295 amino acids with a calculated molecular mass of 37332.76 Da and an estimated isoelectric
point of 4.07. Recombinant chitinase protein expressed intracellularly and revealed high expression in P. pastoris
host. The 37 kDa recombinant chitinase protein developed with antigen antibody confirmed its expression in P.
pastoris.
Conclusion: Conclusively, T. harzianum derived chitinase gene was successfully over expressed in P. pastoris
where recombinant protein was expressed intracellular in the form of inclusion bodies
Muhammad Saleem Iqbal Khan, Anwar Khan, Olawale Samue, Adeyinka, Iqra Yousaf, Saman Riaz, Bisma Bashir, Muhammad Tariq, Bushra Tabassum. (2019) Molecular cloning and expression of recombinant Trichoderma harzianum chitinase in Pichia pastoris, Advancements in Life Sciences, Volume 7, Issue 3.
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