Enhancement of solubility of recombinant alcohol dehydrogenase from Rhodococcus ruber using predictive tool.
Alcohol dehydrogenase
Predictive tools
Protein expression
Solubility
Journal
World journal of microbiology & biotechnology
ISSN: 1573-0972
Titre abrégé: World J Microbiol Biotechnol
Pays: Germany
ID NLM: 9012472
Informations de publication
Date de publication:
02 Sep 2022
02 Sep 2022
Historique:
received:
09
06
2022
accepted:
26
08
2022
entrez:
2
9
2022
pubmed:
3
9
2022
medline:
8
9
2022
Statut:
epublish
Résumé
Solubility is one of key factors influencing the heterologous production of recombinant proteins in biotechnology. Among many aggregation-prone proteins, alcohol dehydrogenase (ADH-A) from Rhodococcus ruber (in this work abbreviated RrADH) shows a great potential in processes involved in the biotransformation of natural compounds. As ADH-A is a potentially high value asset in industrial biotransformation processes, improvement of its solubility would be of major commercial benefit. Predictive tools and in silico analysis provide a fast means for improving protein properties, for selecting appropriate changes, and ultimately for saving costs. We have therefore focused on enhancement of the solubility of RrADH using an online accesible predictive tool Aggrescan 3D 2.0. Selected mutations were introduced into the protein amino acid sequence by using site-directed PCR. This led to a 17% increase in the protein solubility of RrADHmut1 and a 98% increase for RrADHmut2. Moreover, the basic kinetics of the enzyme reaction were positively affected, further optimizing the overall performance of the production process.
Identifiants
pubmed: 36053335
doi: 10.1007/s11274-022-03403-4
pii: 10.1007/s11274-022-03403-4
doi:
Substances chimiques
Recombinant Proteins
0
Alcohol Dehydrogenase
EC 1.1.1.1
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
214Informations de copyright
© 2022. The Author(s), under exclusive licence to Springer Nature B.V.
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