Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Sep;54(3):1623-1633.
doi: 10.1007/s42770-023-01079-y. Epub 2023 Aug 2.

Production, purification, and characterization of cold-active lipase from the psychrotroph Pseudomonas sp. A6

Bahaa Abdella  1 , Asmaa Mohamed Youssif  2 , Soraya A Sabry  2 , Hanan A Ghozlan  3
Affiliations

Production, purification, and characterization of cold-active lipase from the psychrotroph Pseudomonas sp. A6

Bahaa Abdella et al. Braz J Microbiol. 2023 Sep.

Abstract

Cold-active lipases are presently employed extensively in the detergent, chemical intermediate, fine chemical, food, and pharmaceutical industries. Seven cold-adaptive bacteria were isolated from the Mediterranean Sea near Alexandria, Egypt, and tested for their ability to produce cold-active lipase, with the highest activity at 10 °C. The most potent isolate was Pseudomonas sp. A6. To determine the most important variables, the bacterium was exposed to a necessary medium component and environmental factor screening using a single factor-at-a-time approach, followed by a multifactorial Plackett-Burman design strategy. After purification and characterization, the optimal activity levels for the cold-active lipase were figured out. Inoculation of Pseudomonas A6 under near optimum conditions using medium consisting of (g/L) peptone 7.14; soybean oil 7.5% (v/v); K2HPO4, 0.4; MgSO4, 0.1; glucose 2; pH 8; and temperature 10 °C led to a maximum lipase activity anticipated to be 23.36 U/mL. Purified lipase showed the best activity and thermal stability at a pH of 8 and a temperature of 10 °C. The Pseudomonas A6 lipase tolerated the monovalent ions, while greater valence ions did not.

Keywords: Cold-adapted; Lipase; Optimization; Plackett-Burman; Psychrotolerant.

PubMed Disclaimer

Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
Phylogenetic relatedness of isolate A6. The tree was constructed using MEGA X with the closely related representatives of Pseudomonas species. E. coli is included as an outgroup. The tree was drawn to a scale showing the phylogenetic relationship among other Pseudomonas species
Fig. 2
Fig. 2
The effect of different nitrogen sources on the growth and lipase production. Error bars represent the standard error of mean (SEM) of the replica (n=3)
Fig. 3
Fig. 3
The effect of different oils and carbon sources on growth and lipase production. Error bars represent the standard error of mean (SEM) of the replica (n=3)
Fig. 4
Fig. 4
Elucidation of cultivation factors affecting Pseudomonas sp. A6 lipase production using Plackett-Burman experimental design
Fig. 5
Fig. 5
Verification experiments of the applied Plackett-Burman statistical design by comparing the lipase activity produced by Pseudomonas sp. A6 growing on the resulting optimized medium (OP.M), the basal medium (BM), and the anti-optimized medium (A.OP.M). Error bars represent the SEM of three replicas
Fig. 6
Fig. 6
Pseudomonas sp. A6 lipase activity of different fractions obtained using Sephadex G-100
Fig. 7
Fig. 7
Effect of temperature and stability (a), different pHs (b), incubation time (c), and substrate concentration (d) on Pseudomonas sp. A6 purified cold-active lipase activity. Error bars represent the SEM of the replica (n=3)
Fig. 8
Fig. 8
Determination of Km expression of lipase activity
Fig. 9
Fig. 9
SDS-PAGE of purified lipase from Pseudomonas sp. A6: (a) protein marker, (b) crude enzyme, (c) partially purified enzyme by 60% ammonium sulfate, and (d) purified lipase
See this image and copyright information in PMC

References

    1. Kavitha M, Shanthi C. Isolation and characterization of cold active lipase producing Pseudomonas sp. 4 from marine samples of Tamilnadu Coast. Res J Biotechnol. 2013;8:57–62.
    1. Abd-Elnaby HM, Abou-Elela GM, Hussein H, Ghozlan HA, Sabry SA. Characterization and bioremediation potential of marine psychrotolerant Pseudomonas spp. isolated from the Mediterranean Sea, Egypt. Egypt J Aquat Biol Fish. 2019;23(4):669–683. doi: 10.21608/ejabf.2019.63537. - DOI
    1. Gupta R, Gupta N, Rathi P. Bacterial lipases: an overview of production, purification and biochemical properties. Appl Microbiol Biotechnol. 2004;64(6):763–781. doi: 10.1007/s00253-004-1568-8. - DOI - PubMed
    1. Salwoom L, Raja Abd Rahman R, Salleh A, Mohd Shariff F, Convey P, Pearce D, Mohamad Ali M. Isolation, characterisation, and lipase production of a cold-adapted bacterial strain Pseudomonas sp. LSK25 isolated from Signy Island, Antarctica. Molecules. 2019;24(4):715. doi: 10.3390/molecules24040715. - DOI - PMC - PubMed
    1. Kumar A, Mukhia S, Kumar N, Acharya V, Kumar S, Kumar R (2020) A broad temperature active lipase purified from a psychrotrophic bacterium of Sikkim Himalaya with potential application in detergent formulation. Front Bioeng Biotechnol 8. 10.3389/fbioe.2020.00642 - PMC - PubMed

LinkOut - more resources