Academic Journal
Collagenase from Lysinibacillus sphaericus: Production, Optimization and Application in Adipose-Derived Stem Cell Isolation.
| Τίτλος: | Collagenase from Lysinibacillus sphaericus: Production, Optimization and Application in Adipose-Derived Stem Cell Isolation. |
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| Συγγραφείς: | Wang ZZ; School of Life Sciences and Health Engineering, Jiangnan University, Wuxi, 214122, PR China., Xue D; Wuxi Children's Hospital, Wuxi, 214000, PR China. 337855521@qq.com., Qian JY; School of Life Sciences and Health Engineering, Jiangnan University, Wuxi, 214122, PR China. jackieqian@163.com., Wang K; School of Life Sciences and Health Engineering, Jiangnan University, Wuxi, 214122, PR China., Xu LF; School of Life Sciences and Health Engineering, Jiangnan University, Wuxi, 214122, PR China., Ma XD; Cytori Therapeutics LLC, Shanghai, 201802, PR China., Xie N; Cytori Therapeutics LLC, Shanghai, 201802, PR China., Su C; School of Life Sciences and Health Engineering, Jiangnan University, Wuxi, 214122, PR China., Gong JS; School of Life Sciences and Health Engineering, Jiangnan University, Wuxi, 214122, PR China., Xu ZH; College of Biomass Science and Engineering, Sichuan University, Chengdu, 610065, PR China., Shi JS; School of Life Sciences and Health Engineering, Jiangnan University, Wuxi, 214122, PR China. |
| Πηγή: | Applied biochemistry and biotechnology [Appl Biochem Biotechnol] 2026 Oct; Vol. 198 (10), pp. 7448-7463. Date of Electronic Publication: 2026 Jun 13. |
| Τύπος έκδοσης: | Journal Article |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | Publisher: Humana Press Country of Publication: United States NLM ID: 8208561 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1559-0291 (Electronic) Linking ISSN: 02732289 NLM ISO Abbreviation: Appl Biochem Biotechnol Subsets: MEDLINE |
| Imprint Name(s): | Original Publication: Clifton, N.J. : Humana Press, c1981- |
| Ιατρικοί όροι (MeSH): | Bacillaceae*/enzymology , Bacillaceae*/genetics , Stem Cells*/cytology , Collagenases*/biosynthesis , Collagenases*/isolation & purification , Collagenases*/chemistry , Collagenases*/genetics , Cell Separation*/methods , Adipose Tissue*/cytology, Bacillus subtilis/genetics ; Hydrogen-Ion Concentration |
| Περίληψη: | Collagenase, predominantly found in microorganisms, has significant potential for tissue separation and primary cell extraction. In this study, a collagenase Col51 derived from Lysinibacillus sphaericus JN-51 was isolated from sludge samples collected from aquaculture markets and slaughterhouses. Col51 was expressed in Bacillus subtilis (SCK6) and purified using a Ni-NTA column. Enzymatic characterization revealed that the enzyme exhibits optimal activity at 37 °C and pH 7.5. The presence of 10 mM Mg²⁺ and Ca2⁺ enhances Col51 activity, whereas Zn²⁺ and Al³⁺ exert inhibitory effects. Through signal peptide optimization, Col51 expression increased from 53.5 U/mL to 164.8 U/mL in shake-flask experiments. Batch fermentation in a 7-L fermenter further enhanced enzyme activity, reaching a peak of 246.5 U/mL. In adipose stem cell isolation applications, Col51 demonstrated comparable separation efficiency to commercial collagenase I, with cell viability exceeding 90%, and cells exhibiting good morphology and stable growth characteristics. This study successfully identifies and optimizes Col51 collagenase from the non-pathogenic strain Lysinibacillus sphaericus, providing a novel tool enzyme for adipose stem cell isolation. (© 2026. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.) |
| Competing Interests: | Declarations. Ethics Approval: This study was conducted in accordance with the Guidelines for the Euthanasia of Animals (2020 Edition, American Veterinary Medical Association, AVMA) and the ethical standards approved by the Laboratory Animal Center of Jiangnan University, reference number JN.No20250315b1040425[111]. This study utilized SPF-grade male Balb/c mice that were 8 weeks of age and weighed 18–25 g. All procedures were performed to minimize animal distress and suffering. Competing interests: The authors declare that they have no competing interests. |
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| Grant Information: | 2023YFA0914500 National Key Research and Development Program of China; 32301283 National Natural Science Foundation of China |
| Contributed Indexing: | Keywords: Bacillus subtilis; Lysinibacillus sphaericus; Adipose-derived stem cells; Collagenase |
| Substance Nomenclature: | EC 3.4.24.- (Collagenases) |
| SCR Organism: | Lysinibacillus sphaericus |
| Entry Date(s): | Date Created: 20260613 Date Completed: 20260923 Latest Revision: 20260923 |
| Update Code: | 20260924 |
| DOI: | 10.1007/s12010-026-05761-3 |
| PMID: | 42287536 |
| Βάση Δεδομένων: | MEDLINE |
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