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Structural-genetic insight and optimization of protease production from a novel strain of Aeromonas veronii CMF, a gut isolate of Chrysomya megacephala

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Abstract

Structural-genetic characterization of protease producing genes and enzymes from microbial sources are seldom appreciated despite having its substantial utilization in protein engineering or genetic manipulation for biotechnological applications. Aeromonas veronii CMF, a mesophilic bacterium isolated from the gut of Chrysomya megacephala, was found to exhibited significant level of protease activity. For the revelation of genetic potential in relation to protease production, whole genome of this organism was sequenced and analysed while structure–function of different protease enzyme was predicated using various in silico analysis. The 4.5 mb CMF genome was found to encompass various types of protease and mostly they are neutral in nature. Enzyme production was highest in an optimum pH and temperature of 6.0 (32.09 ± 1.015 U/ml) and 35ºC (41.65 ± 1.152 U/ml), respectively. Other culture parameters for optimum production of protease were determined to be inoculum size (1%), incubation period (72 h), shaking condition (125 rpm), carbon and nitrogen source [2% lactose (92.21 ± 3.16 U/ml) and 0.5% urea (163.62 ± 4.31 U/ml), respectively] and effect of surfactants [0.02 mg/ml Tween 80 (174.72 ± 4.48 U/ml)]. Furthermore, A. veronii CMF exhibited significant enzyme production like serine protease (15.22 ± 0.563 U/ml), aspartate protease (33.16 ± 0.762 U/ml) and collagenase (17.26 ± 0.626 U/ml). Genomic information and results of physio-biochemical assays indicate its cost-effective potential use in different enzyme-industry.

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source and surfactants on protease production by CMF

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Acknowledgements

We thank Dr. Wriddhiman Ghosh (Bose Institute, India) for his constructive and intellectual suggestions in manuscript preparation and for providing all kind of computational facilities. Financial support for conducting the microbiological studies was provided by Visva Bharati University via departmental revenue budget grants. R.B. and J.S. received fellowships from Council of Scientific and Industrial Research, GoI. S.B is thankful to Department of Biotechnology, Govt. of India for granting DBT Twinning Project [No. BT/PR25738/NER/95/1329/2017 dated December 24, 2018].

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  1. Rashmi Bhattacherjee, Subhrangshu Mandal and Sandipan Banerjee equal contributions and Joint first author.

Authors and Affiliations

  1. Diptera Section. Zoological Survey of India, M-Block, New Alipore, Kolkata, 700053, India

    Rashmi Bhattacherjee & Dhriti Banerjee

  2. Department of Microbiology, Bose Institute, P-1/12 CIT Scheme VIIM, Kolkata, 700054, India

    Subhrangshu Mandal & Jagannath Sarkar

  3. Mycology and Plant Pathology Laboratory, Department of Botany, Visva Bharati, Siksha Bhavana, Birbhum, Santiniketan, 731235, West Bengal, India

    Sandipan Banerjee, Kunal Kumar Saha & Narayan Chandra Mandal

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Contributions

NCM conceived the study, designed the experiments, interpreted the results and wrote the paper in conjunction with SM. Microbiological experiments and data analysis performed by RB, SM, SB, KS, JS and DB. Bioinformatics analysis performed by SM while SB and KS performed all biochemical experiments. All authors read and vetted the manuscript.

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Correspondence to Narayan Chandra Mandal.

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Bhattacherjee, R., Mandal, S., Banerjee, S. et al. Structural-genetic insight and optimization of protease production from a novel strain of Aeromonas veronii CMF, a gut isolate of Chrysomya megacephala. Arch Microbiol 203, 2961–2977 (2021). https://doi.org/10.1007/s00203-021-02282-x

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