Abstract
Pyruvate carboxylase is a major enzyme for biosynthesis of organic acids like; citric acid, fumeric acid, and l-malic acid. These organic acids play very important role for biological remediation of heavy metals. In this study, gene walking method was used to clone and characterize pyruvate carboxylase gene (F3PYC) from heavy metal resistant indigenous fungal isolate Aspergillus flavus (F3). 3579 bp of an open reading frame which encodes 1193 amino acid protein (isoelectric point: 6.10) with a calculated molecular weight of 131.2008 kDa was characterized. Deduced protein showed 90–95% similarity to those deduced from PYC gene from different fungal strains including; Aspergillus parasiticus, Neosartorya fischeri, Aspergillus fumigatus, Aspergillus clavatus, and Aspergillus niger. Protein generated from the PYC gene was a homotetramer (α4) and having four potential N-linked glycosylation sites and had no signal peptide. Amongst most possible N-glycosylation sites were –N–S–S–I– at 36 amino acid, –N–G–T–V– at 237 amino acid, N–G–S–S– at 517 amino acid, and N–T–S–R– at 1111 amino acid, with several functions have been proposed for the carbohydrate moiety such as thermal stability, pH, and temperature optima for activity and stabilization of the three-dimensional structure. Hence, cloning of F3PYC gene from A. flavus has important biotechnological applications.
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This work was supported by the State Key Laboratory of Environmental Criteria and Risk Assessment (No. SKLECRA2013FP12) and the Public science and technology research funds projects of ocean, China (201105020).
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Dear Editor, this is to certify that the reported work in this manuscript entitled “Cloning and characterization of F3PYC gene encoding Pyruvate carboxylase in Aspergillus flavus (F3) strain” submitted for publication is an original one and has not been submitted for publication elsewhere. We further certify that paper citation to the previously reported work have been given and no data/table/figure has been quoted verbatim from other publications without giving due acknowledgment and without the permission of the authors. The consent of all the authors of this paper has been obtained for submitting the paper to the “3Biotech Journal”.
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Qayyum, S., Khan, I., Bhatti, Z.A. et al. Cloning and characterization of F3PYC gene encoding pyruvate carboxylase in Aspergillus flavus strain (F3). 3 Biotech 7, 245 (2017). https://doi.org/10.1007/s13205-017-0806-6
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DOI: https://doi.org/10.1007/s13205-017-0806-6