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Acinetobacter indicus CCS-12: A New Bacterial Source for the Production and Biochemical Characterization of Thermostable Chitinase with Promising Antifungal Activity

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Abstract

Background

In the recent epoch, chitinolytic enzymes have attained much attention as promising biocontrol agents and ability to convert recalcitrant chitinous wastes into valuable products. Some chitinolytic microorganisms have emerged as the potential candidate to carry a noticeable ability to degrade this polysaccharide in nature.

Objective

Therefore, in this study, a new bacterial strain Acinetobacter indicus CCS-12 was isolated, identified and characterized for the production of a highly active chitinolytic enzyme. The production of extracellularly secreted chitinase was enhanced through the optimization of various cultivation parameters.

Results

Hence, the yield was improved by 2.32-fold in 3ZYB medium (pH 7.0) containing 1% (w/v) colloidal chitin and malt extract when incubated at 45 °C for 48 h with agitation (200 rpm). The enzyme was purified to homogeneity with an estimated molecular mass of 50 kDa by SDS-PAGE and showed optimal activity at pH 7.0 (phosphate buffer) and 60 °C. It was quite stable for 4 h over a range of pH (5.5 to 10.0) and temperature (20 to 60 °C), with a thermal denaturing half-life of 4 h at 75 °C. No obvious inhibitory influence was perceived with different chemical modulators, though enzyme activity was significantly enhanced with Ca2+ (191.76%), Mn2+ (161.25%), Mg2+ (158.44%), Na+ (147.44%), Fe2+ (131.41%) and Cu2+ (125.06%), EDTA (126.02%) and β-mercaptoethanol (131.06%). The Km, Vmax, kcat, and kcat Km−1 values of purified chitinase for colloidal chitin were 0.61 ± 0.11 mg mL−1 and 314.26 ± 0.16 µmol mg−1 min−1, 966.23 s−1 and 1583.98 mL mg−1 s−1, respectively. The purified chitinase exhibited perceptible antifungal activity against some destructive phytopathogens.

Conclusion

With these auspicious features, this enzyme can be effectively used as a biocontrol agent, food additive, for the recycling of chitinous waste materials, in pharmaceutical and various other industries.

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Data Availability

All data generated or analysed during this study are included in this published article [and its supplementary information file].

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Authors and Affiliations

  1. Institute of Industrial Biotechnology, Government College University, Lahore, 54000, Pakistan

    Fatima Akram, Ikram ul Haq & Rabia Akram

  2. Pakistan Academy of Science, Islamabad, Pakistan

    Ikram ul Haq

  3. University of the Punjab, Quaid-i-Azam Campus, Lahore, 54000, Pakistan

    Ayesha Roohi

  4. Kinnaird College for Women, 93 Jail Road, Lahore, 54000, Pakistan

    Ayesha Roohi

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  1. Fatima Akram
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Correspondence to Fatima Akram.

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Akram, F., Haq, I.u., Roohi, A. et al. Acinetobacter indicus CCS-12: A New Bacterial Source for the Production and Biochemical Characterization of Thermostable Chitinase with Promising Antifungal Activity. Waste Biomass Valor 13, 3371–3388 (2022). https://doi.org/10.1007/s12649-022-01753-6

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