Cloning and Co-Expression of Lipase and its Specific Foldase from Ralstonia pickettii BK6
| Autor: | Muhamad Azwar Syah, Andreas Adhi Satya, Esti Puspitasari, Antonius Suwanto, Nisa Rachmania Mubarik |
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| Jazyk: | English<br />Indonesian |
| Rok vydání: | 2022 |
| Předmět: | |
| Zdroj: | Hayati Journal of Biosciences, Vol 30, Iss 1 (2022) |
| Druh dokumentu: | article |
| ISSN: | 1978-3019 2086-4094 |
| DOI: | 10.4308/hjb.30.1.71-80 |
| Popis: | 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. |
| Databáze: | Directory of Open Access Journals |
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