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Cited by 26Enzyme Production and CharacterizationBacterial Genetics and BiotechnologyMicrobial Natural Products and Biosynthesis

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Isolation of lysozyme producing Bacillus subtilis Strains, identification of the new strain Bacillus subtilis BSN314 with the highest enzyme production capacity and optimization of culture conditions for maximum lysozyme production

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Naveed, Muhammad, Tianying, He, Wang, Fenghuan, Yin, Xian, Chan, Malik Wajid Hussain, Ullah, Asad, Xu, Baocai, Aslam, Sadar, Ali, Nawazish, Abbas, Qamar, Hussain, Ishtiaq, Khan, Ali, Khan, Abdul Majeed·2022·Current Research in Biotechnology
DOI 10.1016/j.crbiot.2022.06.002
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Abstract

Lysozyme is a naturally occurring enzyme that is ubiquitous and plays an important role in innate immunity through its potent bacteriolytic activity. The main objective of this study was to isolate, screen and identify lysozyme-producing Bacillus subtilis strains by 16S rDNA sequencing. Of the four Bacillus subtilis strains isolated (B18, B66, B70, and B124), only one (B66) was selected for further study because it exhibited the comparatively highest enzymatic activity. The 16S rDNA sequence of strain B66 was submitted to the Genbank database with accession number OM327451 and was assigned the name Bacillus subtilis BSN314. Bacillus subtilis BSN314 was selected for further study to maximize biomass production. To obtain the maximum biomass product of Bacillus subtilis BSN314, two statistical methods were applied, one related to the nutritional conditions affecting the growth medium (chemical components), called the single factor optimization method, and the other showing their effects on the cultivation conditions (orthogonal method). The results showed that the composition of the culture medium (carbon, nitrogen, inorganic salts and surfactants) and the culture conditions (temperature, pH, shaking speed and percentage of inoculum) have a specific influence on the biomass production of Bacillus subtilis BSN314. The culture medium containing the combination of A2B3C1D2 with 1.5% glucose, 2.5% soybean peptone, 0.5% K2HPO4, and 0.1% Tween-20 was determined to be the optimal medium for biomass production whereas, the order of culture media based on the effects of the selected conditions having lysozyme activity is as follows; glucose % > soyapeptone % > Tween-20 % > K2HPO4 %. The results showed that the A2B3C1D2 ratio had the highest enzyme activity of 55 U/mL.

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CC-BY-NC-ND · From the PDF
Fig. 1 Colonies and transparent circles on A and B plates, were obtained by preliminary screening from soil samples by double-layer method.
Fig. 1 Colonies and transparent circles on A and B plates, were obtained by preliminary screening from soil samples by double-layer method.CC-BY-NC-ND · From the PDF
Fig. 2 16S rDNA Electrophoresis Image. Where, M = DNA marker; 1 = B66; 2 = B124; 3 = B18; 4 = B70; 5 = negative control.
Fig. 2 16S rDNA Electrophoresis Image. Where, M = DNA marker; 1 = B66; 2 = B124; 3 = B18; 4 = B70; 5 = negative control.CC-BY-NC-ND · From the PDF
Fig. 3 Effect of carbon (A), and nitrogen (B) sources on lysozyme production.
Fig. 3 Effect of carbon (A), and nitrogen (B) sources on lysozyme production.CC-BY-NC-ND · From the PDF
Image 5
CC-BY-NC-ND · From the PDF
Fig. 5 Effect of temperature (A), and pH (B) on lysozyme production.
Fig. 5 Effect of temperature (A), and pH (B) on lysozyme production.CC-BY-NC-ND · From the PDF
Fig. 6 Effect of shaking speed (A), and inoculum volume (B) on lysozyme production.
Fig. 6 Effect of shaking speed (A), and inoculum volume (B) on lysozyme production.CC-BY-NC-ND · From the PDF

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Authors
Naveed, M., Tianying, H., Wang, F., Yin, X., Chan, M. W. H., Ullah, A., Xu, B., Aslam, S., Ali, N., Abbas, Q., Hussain, I., Khan, A., & Khan, A. M.
Affiliations
  • Beijing Technology and Business University (CN)
  • Federal Urdu University (PK)
  • Pakistan Council of Scientific & Industrial Research (PK)
  • University of Karachi (PK)
  • Karakoram International University (PK)
Corresponding author
  • Wang, Fenghuan
  • Xu, Baocai
ORCID
Venue
Current Research in Biotechnology
Publisher
Elsevier BV
Published
2022
Volume
4
Pages
290-301
ISSN
2590-2628 (ISSN-L)
DOI
10.1016/j.crbiot.2022.06.002
OpenAlex
W4283445989
Cited by
26
References
1
Citations by year
2026: 3 · 2025: 10 · 2024: 7 · 2023: 6
FWCI
2.75
Type
journal article
Language
en
Open access
Licence
Locations
  • Current Research in Biotechnology (publishedVersion, cc-by-nc-nd)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • National Natural Science Foundation of China (22178006)
  • National Key Research and Development Program of China (2021YFC2102800)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Enzyme Production and Characterization
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Immunology and Microbiology › Applied Microbiology and Biotechnology › Microbial Natural Products and Biosynthesis
Keywords
lysozyme production, Bacillus subtilis, bacterial isolation, 16S rRNA identification, culture medium optimization, orthogonal experimental design, biomass production, glucose, Tween 20, culture conditions, enzyme activity

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