Research Article

The lipolytic activity of LipJ, a stress-induced enzyme, is regulated by its C-terminal adenylate cyclase domain

https://orcid.org/0000-0002-4444-3431

Department of Biotechnology, BMS Block-1, South Campus, Panjab University, Chandigarh, India

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Jashandeep Kaur

https://orcid.org/0000-0002-6984-8945

Department of Biotechnology, BMS Block-1, South Campus, Panjab University, Chandigarh, India

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Pratibha Maan

https://orcid.org/0000-0002-2410-5312

Department of Biotechnology, BMS Block-1, South Campus, Panjab University, Chandigarh, India

Department of Experimental Medicine and Biotechnology PGIMER, Chandigarh, India

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Arbind Kumar

COVID Testing Facility, CSIR-Institute of Himalayan Bioresources & Technology, Palampur, Himachal Pradesh, India

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Jagdeep Kaur

*Author for correspondence: Tel.: +91 0172 253 4085;

E-mail Address: jagsekhon@pu.ac.in

Department of Biotechnology, BMS Block-1, South Campus, Panjab University, Chandigarh, India

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Published Online:7 May 2021https://doi.org/10.2217/fmb-2020-0223

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Abstract

Aim: The confirmation of lipolytic activity and role of Rv1900c in the Mycobacterium physiology Methods:rv1900c/N-terminus domain (rv1900NT) were cloned in pET28a/Escherichia coli, purified by affinity chromatography and characterized. Results: A zone of clearance on tributyrin-agar and activity with pNP-decanoate confirmed the lipolytic activity of Rv1900c. The Rv1900NT demonstrated higher enzyme specific activity, V_max and k_cat, but Rv1900c was more thermostable. The lipolytic activity of Rv1900c decreased in presence of ATP. Mycobacterium smegmatis expressed rv1900c/rv1900NT-altered colony morphology, growth, cell surface properties and survival under stress conditions. The effect was more prominent with Rv1900NT as compared with Rv1900c. Conclusion: The study confirmed the lipolytic activity of Rv1900c and suggested its regulation by the adenylate cyclase domain and role in the intracellular survival of bacteria.

Lay abstract

Tuberculosis (TB) remains the top contagious/infectious killer in the world. It is caused by the bacteria Mycobacterium tuberculosis. The bacteria resides/replicates in the immune cell that normally has to eradicate infectious microorganisms. Though the treatment of TB is available, the emergence of drug-resistant bacteria is of major concern. The treatment of drug-resistant TB has been reported to be more difficult due to lengthy and complex treatment regimens. Therefore, there is an urgent need for new and better drugs to treat TB/drug-resistant TB. For this purpose understanding the role of each protein in the physiology of mycobacteria is required. Lipids play a critical role in the intracellular survival of this pathogen in the host. Our study demonstrated that LipJ supported the intracellular survival of bacteria. Therefore, it could be a potential drug target.

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Papers of special note have been highlighted as: • of interest; •• of considerable interest

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Vol. 16, No. 7

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History

Received 31 August 2020

Accepted 24 March 2021

Published online 7 May 2021

Published in print May 2021

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Supplementary data

To view the supplementary data that accompany this paper please visit the journal website at: www.futuremedicine.com/doi/suppl/10.2217/fmb-2020-0223

Author contributions

J Kaur framed the idea, outlined the study, supervised the work and edited the final draft of the manuscript. B Kumari conducted literature searches, accomplished the major experimental work and wrote the first draft of the manuscript. A Kumar assisted in bioinformatics and biophysical analysis, J Kaur and P Maan helped in editing the final draft of the manuscript. All the authors have read and validated the final manuscript.

Financial & competing interests disclosure

B Kumari received fellowship for carrying out her PhD work from CSIR, India. The instruments used for the present study were supported by DST-FIST, UGC-SAP and DST-PURSE grant to the department of Biotechnology. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

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