Crystallization and preliminary X-ray diffraction analysis of<scp>L</scp>-aminoacylase from the hyperthermophilic archaeon<i>Thermococcus litoralis</i>

Hollingsworth, Edward J.; Isupov, Michail N.; Littlechild, Jennifer A.

doi:10.1107/s0907444901021266

Cited by 8 publications

(2 citation statements)

References 21 publications

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“…The L-aminoacylase has been cloned and overexpressed in high yields from E. coli. This has enabled its biochemical characterization and crystallization [6,7]. The recombinant enzyme is thermostable, with a half-life of 25 h at 70 • C and can be immobilized and packed into a column reactor which can be used to convert substrate into product continuously for up to 10 days at 60 • C with no loss in activity of the enzyme [8].…”

Section: Aminoacylasementioning

confidence: 99%

Natural methods of protein stabilization: thermostable biocatalysts

Littlechild

Guy

Connelly

et al. 2007

Biochemical Society Transactions

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Enzymes that are naturally found in thermophilic and hyperthermophilic organisms are being used as robust biocatalysts in the fine chemical and pharmaceutical industries. They have important use in these industries due to their increased stability which is often required during commercial reaction conditions. The approach used in these studies is to learn how nature has managed to stabilize these proteins using a detailed knowledge of their biochemical properties and three-dimensional structures. This is illustrated with several different classes of enzyme that have been studied at Exeter. These include alcohol dehydrogenase, aminoacylase, pyroglutamyl carboxypeptidase, gamma-lactamase, dehalogenase and lysophospholipase.

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Section: Aminoacylasementioning

confidence: 99%

Natural methods of protein stabilization: thermostable biocatalysts

Littlechild

Guy

Connelly

et al. 2007

Biochemical Society Transactions

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“…These ions have been reported to inhibit the proteolytic activities of elastase strain K (Rahman et al, 2011), ME-4 (Cheng et al, 2009) and PseA (Gupta et al, 2005) which also happened to be metalloproteases. The highest half-life of aminoacylase was reported by Hollingsworth (2002), who found that the stability of aminoacylase from Thermococcus litoralis was 25 h at 70°C. Aminoacylase SZN can withstand a broad range of inhibitors and did not exhibit significant effects on its stability in detergent especially Tween 20.…”

Section: Discussionmentioning

confidence: 94%

Characterization of thermostable aminoacylase from Geobacillus sp. strain SZN

Adenan

Wong

Zain

et al. 2019

APJMBB

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Aminoacylase (EC 3.5.1.14) hydrolyzes N–acetylated amino acids to produce amino acids. Although thermostable aminoacylase has been commercially produced since 2004, there was a knowledge gap in the field of understanding aminoacylase thermostability from a structural point of view. This study investigated the physical and structural properties of the purified thermostable aminoacylase SZN. The spectropolarimetry data for structural determination has indicated a gradual decrease of α-helix from 36 to 27.6%, followed by tremendous disorientation of the structure at the transition of temperatures from 60 to 70°C (27.6 to 19.5%). In contrast, the percentage of β-sheet has increased steadily over the tested temperatures. The α-helix, where notable metal binding and catalytic residues are located, was totally weakened at temperatures above 70C, thus resulted in loss of activity. The loss of the α-helical structure could further explain drastic deterioration of activity at temperatures beyond 70C. The activity of aminoacylase SZN was enhanced by divalent metal ions, such as Mn2+ and Cu2+, and inhibited by detergent Triton-X-100. As a conclusion, the isolated aminoacylase SZN was characterized as a thermostable enzyme based on the α-helical structure integrity and functional stability in high temperatures. This enzyme could be used as an alternative enzyme for bioindustries in view of its activity enhancement in high temperatures and stability in various tested inhibitors.

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