Cloning and Co-Expression of Lipase and its Specific Foldase from Ralstonia pickettii BK6
This study aimed to obtain a functional lipase (LipRM) from Ralstonia pickettii BK6 through a co-expression involving its foldase. The ORF of the LipRM was 999 bp while the gene encoding of lipase-specific foldase (LifRM) was 1,030 bp. LipRM and LifRM genes were cloned into a plasmid and were successfully co-expressed in Escherichia coli strains to produce functional LipRM. Enzyme activity from partially purified enzymes showed quite surprising results, LipRM activity in E. coli BL21 (DE3) was 25.84 U/ml, while the other strains (DH5α, HB101, S17-1λpir) were 628.98 U/ml, 761 U/ml, and 1206.46 U/ml, respectively. The highest relative activity of LipRM was found at 50-55°C and pH 7-8 with pNP-laurate (C12) as the preferred substrate specificity. LipRM activity was enhanced sharply in the presence of 30% organic solvents (methanol and ethanol) but decreased by more than 50% in the presence of detergents. This study was the first to report heterologous expression of Ralstonia pickettii lipase employing its native foldase resulting in functional lipase from subfamily I.2.
Copyright (c) 2023 Muhamad Azwar Syah, Andreas Adhi Satya, Esti Puspitasari, Antonius Suwanto, Nisa Rachmania Mubarik
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
HAYATI J Biosci article's license is CC-BY-NC. This license lets others distribute, remix, tweak, and build upon author's work, as long as they credit the original creation.
Authors who submit and publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal/publisher non exclusive publishing rights with the work simultaneously licensed under a https://creativecommons.org/
licenses/by-nc/4.0/Attributi on — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use.
- Authors can still use their work commercially
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).