Genome-Driven Discovery of a Fe<sup>2+</sup>-Dependent Chitin Deacetylase from <i>Bacillus pumilus</i> B866 with Enhanced Thermostability
Chitin deacetylase (CDA) plays a pivotal role in converting chitin to chitosan, yet industrial applications remain constrained by low enzymatic activity, instability under process conditions, and insufficient understanding of metalloenzyme activation mechanisms. Addressing these challenges, we condu...
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| Những tác giả chính: | , , , , , |
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| Định dạng: | Bài viết |
| Ngôn ngữ: | Tiếng Anh |
| Được phát hành: |
MDPI AG
2025-06-01
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| Loạt: | Fermentation |
| Những chủ đề: | |
| Truy cập trực tuyến: | https://www.mdpi.com/2311-5637/11/6/327 |
| Các nhãn: |
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| Tóm tắt: | Chitin deacetylase (CDA) plays a pivotal role in converting chitin to chitosan, yet industrial applications remain constrained by low enzymatic activity, instability under process conditions, and insufficient understanding of metalloenzyme activation mechanisms. Addressing these challenges, we conducted a genome-driven investigation of 151 salt-tolerant <i>Bacillus</i> strains to identify robust CDAs tailored for industrial demands. Genomic analysis revealed 120 strains harboring CDA genes, with <i>Bacillus pumilus</i> B866 exhibiting the highest native activity (105.93 U/mL). Through systematic medium optimization—identifying lactose, yeast extract, and FeSO<sub>4</sub> as critical components—CDA production in B866 surged to 191.32 U/mL, a 2.39-fold increase over baseline. Heterologous expression of <i>Bp</i>CDA in <i>E. coli</i> yielded a recombinant enzyme (123.27 U/mL) with superior thermostability (retaining > 42.9% activity after 24 h at 55 °C) and broad pH adaptability (>81.4% activity at pH 7.0–9.0). Notably, <i>Bp</i>CDA demonstrated unique Fe<sup>2+</sup>-dependent activation (186.4% activity enhancement at 1 mM), contrasting with Mg<sup>2+</sup>-dependent systems in prior studies. Comparative genomic and pan-genome analyses underscored evolutionary adaptations linked to saline–alkaline niches, while biosynthetic gene cluster profiling revealed strain-specific metabolic potentials independent of genome size. This study resolves critical limitations in CDA performance by integrating genome mining, targeted screening, and metalloenzyme engineering, establishing a scalable platform for sustainable chitin valorization. The optimized <i>Bp</i>CDA, with its industrial-compatible stability and novel activation mechanism, represents a significant advancement toward efficient, eco-friendly chitosan production. |
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| số ISSN: | 2311-5637 |