The Effect of N-Terminal Domain Removal towards the Biochemical and Structural Features of a Thermotolerant Lipase from an Antarctic Pseudomonas sp. Strain AMS3

Abstract

Lipase plays an important role in industrial and biotechnological applications. Lipases have been subject to modification at the N and C terminals, allowing better understanding of lipase stability and the discovery of novel properties. A thermotolerant lipase has been isolated from Antarctic Pseudomonas sp. The purified Antarctic AMS3 lipase (native) was found to be stable across a broad range of temperatures and pH levels. The lipase has a partial Glutathione-S-transferase type C (GST-C) domain at the N-terminal not found in other lipases. To understand the influence of N-terminal GST-C domain on the biochemical and structural features of the native lipase, the deletion of the GST-C domain was carried out. The truncated protein was successfully expressed in E. coli BL21(DE3). The molecular weight of truncated AMS3 lipase was approximately ~45 kDa. The number of truncated AMS3 lipase purification folds was higher than native lipase. Various mono and divalent metal ions increased the activity of the AMS3 lipase. The truncated AMS3 lipase demonstrated a similarly broad temperature range, with the pH profile exhibiting higher activity under alkaline conditions. The purified lipase showed a substrate preference for a long carbon chain substrate. In addition, the enzyme activity in organic solvents was enhanced, especially for toluene, Dimethylsulfoxide (DMSO), chloroform and xylene. Molecular simulation revealed that the truncated lipase had increased structural compactness and rigidity as compared to native lipase. Removal of the N terminal GST-C generally improved the lipase biochemical characteristics. This enzyme may be utilized for industrial purposes.

Characterisation of physicochemical properties, flavour components and microbial community in Chinese Guojing roasted sesame-like flavourDaqu

“…Indeed, previous studies on Pseudomonas have reported it to be involved in food spoilage . Some studies showed that Pseudomonas not only produce some important enzymes, but also produce important biological metabolites that are used in industry . It is also worth noting that Pseudomonas can produce antibacterial substances and may play a role in improving the structure of microbial flora in Daqu or Baijiu production .…”

“…Some studies showed that Pseudomonas not only produce some important enzymes, but also produce important biological metabolites that are used in industry (35). It is also worth noting that Pseudomonas can produce antibacterial substances and may play a role in improving the structure of microbial flora in Daqu or Baijiu production (35). Yao et al reported Sphingobium for the first time in Baijiu, and this is the second report of it in Baijiu (6).…”

Immobilization of an Antarctic Pseudomonas AMS8 Lipase for Low Temperature Ethyl Hexanoate Synthesis

“…In contrast with polar organic solvent, it has been reported that polar solvent can distort the water layer around the enzyme which make the enzyme less stable and lead to deactivation of the enzyme [16]. AMS8 lipase showed the maximum ester conversion was only at 36% in 1-butanol, a polar organic solvent which was 1-butanol.…”

Engineering an Alcohol-Forming Fatty Acyl-CoA Reductase for Aldehyde and Hydrocarbon Biosynthesis in Saccharomyces cerevisiae

“…As predicted, no FALDR and FACR activities were detected from the C- and N-terminal domains, respectively. It is unclear why, relative to wild-type maFACR, maFACR Nter has higher FALDR activity while maFACR Cter has lower FACR activity, but it is known that protein truncation can alter structural flexibility, substrate accessibility, and quaternary structure of enzymes, which could all affect the activity ( Gorfe et al, 2009 ; Yu et al, 2017 ; Latip et al, 2018 ). Nonetheless, these results strongly validate the hypothesis that the C-terminal domain is an FACR while the N-terminal domain functions as an FALDR.…”