Heterologous expressing melittin in a probiotic yeast to evaluate its function for promoting NSC-34 regeneration.
Kluyveromyces marxianus
Bee venom
Cell growth–related factor gene expression
Heterologous expression system
Melittin
NSC-34 motor neurons
Journal
Applied microbiology and biotechnology
ISSN: 1432-0614
Titre abrégé: Appl Microbiol Biotechnol
Pays: Germany
ID NLM: 8406612
Informations de publication
Date de publication:
28 Oct 2024
28 Oct 2024
Historique:
received:
21
03
2024
accepted:
16
10
2024
revised:
21
09
2024
medline:
28
10
2024
pubmed:
28
10
2024
entrez:
28
10
2024
Statut:
epublish
Résumé
Melittin is a bioactive peptide and the predominant component in bee venom (BV), studied for its many medical properties, such as antibacterial, anti-inflammatory, anti-arthritis, nerve damage reduction, and muscle cell regeneration. Melittin is primarily obtained through natural extraction and chemical synthesis; however, both methods have limitations and cannot be used for mass production. This study established a heterologous melittin expression system in the probiotic yeast Kluyveromyces marxianus. This yeast was selected for its advantages in stress tolerance and high secreted protein yields, simplifying purification. A > 95% high-purity melittin (MET) and its precursor promelittin (ProMET) were successfully produced and purified at 1.68 μg/mL and 3.33 μg/mL concentrations and verified through HPLC and mass spectrum. The functional test of the NSC-34 cell regeneration revealed that MET achieved the best activity compared to ProMET and the natural-extracted BV groups. Growth-related gene expressions were evaluated, including microtubule-associated protein 2 (MAP2), microtubule-associated protein Tau (MAPT), growth-associated protein 43 (GAP-43), choline acetyltransferase (ChAT), vesicular acetylcholine transporter (VAChT), and acetylcholine esterase (AChE). The results indicated that treating MET increased MAP2, GAP-43, and VAChT expressions, in which cholinergic signaling is related to neurological functions. A heterologously expressed melittin in a probiotic yeast and its potential for promoting NSC-34 regeneration described here facilitate commercial and therapeutic use. KEY POINTS: • MET and its precursor ProMET were successfully hetero-expressed in K. marxianus • > 95% high-purity MET and ProMET were purified at 1.68 μg/mL and 3.33 μg/mL • MET has no cytotoxicity toward NSC-34 and significantly promotes NSC-34 growth.
Identifiants
pubmed: 39466458
doi: 10.1007/s00253-024-13336-7
pii: 10.1007/s00253-024-13336-7
doi:
Substances chimiques
Melitten
20449-79-0
Recombinant Proteins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
496Subventions
Organisme : National Science and Technology Council
ID : NSTC 111-2313-B-005-037-
Organisme : National Science and Technology Council
ID : NSTC 112-2313-B-005-048-
Organisme : National Science and Technology Council
ID : NSTC 113-2313-B-005-015-
Organisme : Tungs' Taichung MetroHarbor Hospital
ID : TTMHH-NCHULS111002
Organisme : Tungs' Taichung MetroHarbor Hospital
ID : TTMHH-NCHULS112003
Organisme : Tungs' Taichung MetroHarbor Hospital
ID : TTMHH-NCHULS113003
Informations de copyright
© 2024. The Author(s).
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