Effect and mechanism of signal peptide and maltose on recombinant type III collagen production in Pichia pastoris.
Anti-degradation
Maltose mechanism
Pichia pastoris GS115
Recombinant type III collagen production
Transcriptome
α-Factor peptide
Journal
Applied microbiology and biotechnology
ISSN: 1432-0614
Titre abrégé: Appl Microbiol Biotechnol
Pays: Germany
ID NLM: 8406612
Informations de publication
Date de publication:
Jul 2023
Jul 2023
Historique:
received:
22
02
2023
accepted:
09
05
2023
revised:
04
05
2023
medline:
3
7
2023
pubmed:
18
5
2023
entrez:
18
5
2023
Statut:
ppublish
Résumé
Recombinant type III collagen plays an important role in cosmetics, wound healing, and tissue engineering. Thus, increasing its production is necessary. After an initial increase in output by modifying the signal peptide, we showed that adding 1% maltose directly to the medium increased the yield and reduced the degradation of recombinant type III collagen. We initially verified that Pichia pastoris GS115 can metabolize and utilize maltose. Interestingly, maltose metabolism-associated proteins in Pichia pastoris GS115 have not yet been identified. RNA sequencing and transmission electron microscopy were performed to clarify the specific mechanism of maltose influence. The results showed that maltose significantly improved the metabolism of methanol, thiamine, riboflavin, arginine, and proline. After adding maltose, the cell microstructures tended more toward the normal. Adding maltose also contributed to yeast homeostasis and methanol tolerance. Finally, adding maltose resulted in the downregulation of aspartic protease YPS1 and a decrease in yeast mortality, thereby slowing down recombinant type III collagen degradation. KEY POINTS: • Co-feeding of maltose improves recombinant type III collagen production. • Maltose incorporation enhances methanol metabolism and antioxidant capacity. • Maltose addition contributes to Pichia pastoris GS115 homeostasis.
Identifiants
pubmed: 37199749
doi: 10.1007/s00253-023-12579-0
pii: 10.1007/s00253-023-12579-0
doi:
Substances chimiques
Recombinant Proteins
0
Collagen Type III
0
Maltose
69-79-4
Protein Sorting Signals
0
Methanol
Y4S76JWI15
YPS1 protein, S cerevisiae
EC 3.4.23.25
Aspartic Acid Endopeptidases
EC 3.4.23.-
Saccharomyces cerevisiae Proteins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
4369-4380Subventions
Organisme : National Key R&D Program of China
ID : 2019YFA0905200
Organisme : National Natural Science Foundation of China
ID : 22108229
Organisme : the Xi'an Science and Technology Project
ID : 20GXSF0004
Organisme : Xi'an Science and Technology Project
ID : 20191422315KYPT014JC016
Informations de copyright
© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.
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