title: Prolonged Production and Aggregation Complexity of Cold-Active Lipase from Pseudomonas proteolytica (GBPI_Hb61) Isolated from Cold Desert Himalaya.
abstract: Pseudomonas, being the common inhabitant of colder environments, are suitable for the production of cold-active enzymes. In the present study, a newly isolated strain of Pseudomonas from cold desert site in Indian Himalayan Region, was investigated for the production of cold-active lipase. The bacteria were identified as Pseudomonas proteolytica by 16S rDNA sequencing. Lipase production by bacteria was confirmed by qualitative assay using tributyrin and rhodamine-B agar plate method. The bacterium produced maximum lipase at 25 °C followed by production at 15 °C while utilizing olive, corn, as well as soybean oil as substrate in lipase production broth. Enzyme produced by bacteria was partially purified using ammonium sulphate fractionation. GBPI_Hb61 showed aggregation behaviour which was confirmed using several techniques including gel filtration chromatography, dynamic light scattering, and native PAGE. Molecular weight determined by SDS-PAGE followed by in-gel activity suggested two lipases of nearly similar molecular weight of ~50 kDa. The enzyme showed stability in wide range of pH from 5 to 11 and temperature up to 50 °C. The enzyme from GBPI_Hb61 exhibited maximum activity toward p-nitrophenyldecanoate (C10). The stability of enzyme was not affected with methanol while it retained more than 75% activity when incubated with ethanol, acetone, and hexane. The bacterium is likely to be a potential source for production of cold-active lipase with efficient applicability under multiple conditions.
authors.affiliation: Biotechnological Applications, G B Pant National Institute of Himalayan Environment and Sustainable Development, Kosi-Katarmal, Almora, Uttarakhand, 263 643, India.
authors.forename: Rahul
authors.initials: R
authors.lastname: Jain
authors.affiliation: Biotechnological Applications, G B Pant National Institute of Himalayan Environment and Sustainable Development, Kosi-Katarmal, Almora, Uttarakhand, 263 643, India. anita@gbpihed.nic.in.
authors.forename: Anita
authors.initials: A
authors.lastname: Pandey
authors.affiliation: Department of Microbiology, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, 226 031, India.
authors.forename: Mukesh
authors.initials: M
authors.lastname: Pasupuleti
authors.affiliation: Department of Biotechnology, Kumaun University, Bhimtal Campus, Bhimtal, Uttarakhand, 263 136, India.
authors.forename: Veena
authors.initials: V
authors.lastname: Pande
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D003081:Cold Climate; D004789:Enzyme Activation; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000378:metabolism*; D008970:Molecular Weight; D011549:Pseudomonas / Q000201:enzymology* / Q000302:isolation & purification*; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D008049:Lipase
keywords: Aggregation; Cold-active lipase; Indian Himalayan Region; Pseudomonas