Appraisal of cytotoxicity and acrylamide mitigation potential of L-asparaginase SlpA from fish gut microbiome.

Autor: Johny TK; Department of Biotechnology, Cochin University of Science and Technology, Cochin, Kerala, 682022, India., Puthusseri RM; Department of Biotechnology, Cochin University of Science and Technology, Cochin, Kerala, 682022, India., Saidumohamed BE; Department of Biotechnology, Cochin University of Science and Technology, Cochin, Kerala, 682022, India., Sheela UB; Department of Plant Biotechnology, College of Agriculture, Vellayani, Kerala, 695522, India., Puthusseri SP; Department of Biotechnology, Cochin University of Science and Technology, Cochin, Kerala, 682022, India., Sasidharan RS; Department of Zoology, Government College Kariavattom, University of Kerala, Thiruvananthapuram, Kerala, 695581, India., Bhat SG; Department of Biotechnology, Cochin University of Science and Technology, Cochin, Kerala, 682022, India. saritagbhat@gmail.com.
Jazyk: angličtina
Zdroj: Applied microbiology and biotechnology [Appl Microbiol Biotechnol] 2022 May; Vol. 106 (9-10), pp. 3583-3598. Date of Electronic Publication: 2022 May 17.
DOI: 10.1007/s00253-022-11954-7
Abstrakt: L-asparaginase catalyzes the hydrolysis of L-asparagine to L-aspartic acid and ammonia. It has application in the treatment of acute lymphoblastic leukemia in children, as well as in other malignancies, in addition to its role as a food processing aid for the mitigation of acrylamide formation in the baking industry. Its use in cancer chemotherapy is limited due to problems such as its intrinsic glutaminase activity and associated side effects, leading to an increased interest in the search for novel L-asparaginases without L-glutaminase activity. This study reports the cloning and expression of an L-asparaginase contig obtained from whole metagenome shotgun sequencing of Sardinella longiceps gut microbiota. Purified recombinant glutaminase-free L-asparaginase SlpA was a 74 kDa homodimer, with maximal activity at pH 8 and 30 °C. K m and V max of SlpA were determined to be 3.008 mM and 0.014 mM/min, respectively. SlpA displayed cytotoxic activity against K-562 (chronic myeloid leukemia) and MCF-7 (breast cancer) cell lines with IC 50 values of 0.3443 and 2.692 U/mL, respectively. SlpA did not show any cytotoxic activity against normal lymphocytes and was proved to be hemocompatible. Pre-treatment of biscuit and bread dough with different concentrations of SlpA resulted in a clear, dose-dependent reduction of acrylamide formation during baking. KEY POINTS: • Cloned and expressed L-asparaginase (SlpA) from fish gut microbiota • Purified SlpA displayed good cytotoxicity against K-562 and MCF-7 cell lines • SlpA addition caused a significant reduction of acrylamide formation during baking.
(© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)
Databáze: MEDLINE
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