The purification and characterization of an extremely thermostable alpha-amylase from the hyperthermophilic archaebacterium Pyrococcus furiosus.

Laderman, Kenneth A.; Davis, Bradley R.; Krutzsch, Henry C.; Lewis, Marc S.; Griko, Yuri V.; Privalov, Peter L.; Anfinsen, Christian B.

doi:10.1016/s0021-9258(20)80538-9

Cited by 154 publications

(26 citation statements)

References 32 publications

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“…Several α-specific CAZymes were thus only enriched in the xylan samples. The highest enrichment was observed for EGIDFPOO_00375 from the GH57 family ( Supplementary Figure 2 in Supplementary Data Sheet 1 ) that is homologous to the characterized pullulan hydrolase TK-PUL from T. kodakarensis ( Ahmad et al, 2014 ), 4-α-glucanotransferase from T. litoralis ( Jeon et al, 1997 ) and α-amylase from P. furiosus ( Laderman et al, 1993 ). These enzymes have been reported to hydrolyze multiple α-linked polysaccharides, including starch, glycogen, dextrin, amylose, amylopectin, and different cyclodextrins.…”

Section: Resultsmentioning

confidence: 99%

Activity-Based Protein Profiling for the Identification of Novel Carbohydrate-Active Enzymes Involved in Xylan Degradation in the Hyperthermophilic Euryarchaeon Thermococcus sp. Strain 2319x1E

et al. 2022

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Activity-based protein profiling (ABPP) has so far scarcely been applied in Archaea in general and, especially, in extremophilic organisms. We herein isolated a novel Thermococcus strain designated sp. strain 2319x1E derived from the same enrichment culture as the recently reported Thermococcus sp. strain 2319x1. Both strains are able to grow with xylan as the sole carbon and energy source, and for Thermococcus sp. strain 2319x1E (optimal growth at 85°C, pH 6–7), the induction of xylanolytic activity in the presence of xylan was demonstrated. Since the solely sequence-based identification of xylanolytic enzymes is hardly possible, we established a complementary approach by conducting comparative full proteome analysis in combination with ABPP using α- or β-glycosidase selective probes and subsequent mass spectrometry (MS)-based analysis. This complementary proteomics approach in combination with recombinant protein expression and classical enzyme characterization enabled the identification of a novel bifunctional maltose-forming α-amylase and deacetylase (EGDIFPOO_00674) belonging to the GH57 family and a promiscuous β-glycosidase (EGIDFPOO_00532) with β-xylosidase activity. We thereby further substantiated the general applicability of ABPP in archaea and expanded the ABPP repertoire for the identification of glycoside hydrolases in hyperthermophiles.

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Section: Resultsmentioning

confidence: 99%

Activity-Based Protein Profiling for the Identification of Novel Carbohydrate-Active Enzymes Involved in Xylan Degradation in the Hyperthermophilic Euryarchaeon Thermococcus sp. Strain 2319x1E

et al. 2022

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“…Moreover, detection of maltose as a major component from the hydrolyzed product confirmed its nature as β type ( 17 ). It has been reported that most bacilli producing amylase have optimum temperature stability between 40 and 70°C ( 14 , 20 – 22 ). Obi and Odibo ( 17 ) reported that β-amylase withstood up to 70°C, while Fogarty and Griffin ( 23 ) observed temperature tolerance of amylase up to 52°C ( Bacillus polymyxa ) and 70°C from another Bacillus sp.…”

Section: Discussionmentioning

confidence: 99%

Exploration for Thermostable β-Amylase of a Bacillus sp. Isolated from Compost Soil to Degrade Bacterial Biofilm

Pan

2021

Microbiol Spectr

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“…Previous studies showed that some high-temperature α-amylases exhibited excellent thermostability and half-life (T 1/2 ) can reach 4-12 h at their optimum temperature. For example, optimum temperature of α-amylase from Pyrococcus furiosus was 100 °C (Laderman et al 1993), and optimum temperature of α-amylase from B. amyloliquifaciens TSWK1-1 was 70 °C (Kikani and Singh 2011). Despite different optimum temperatures, T 1/2 of both α-amylases reached 12 h at their optimum temperature, respectively.…”

Section: Introductionmentioning

confidence: 99%

Expression of Bacillus licheniformis α-amylase in Pichia pastoris without antibiotics-resistant gene and effects of glycosylation on the enzymic thermostability

Zhuang

et al. 2019

3 Biotech

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Thermostable α-amylases are widely used in industry. The α-amylase from Bacillus licheniformis (BLA) with six potential glycosylation sites possessed excellent thermal and pH stability and high activity. Here, it was expressed in Pichia pastoris. The Pic-BLA-producing yeast without any antibiotics-resistant gene was cultivated in flasks and the amylase activity in fermentation supernatant reached 900 U/mL. The recombinant α-amylase Pic-BLA produced in P. pastoris was deeply glycosylated with 30% increase in molecular mass (MM). The deglycosylation treatment by Endoglycosidase H (Endo H) reduced the MM of Pic-BLA. Thermostability analysis showed that Pic-BLA and deglycosylated Pic-BLA were similar in heat tolerance. In order to eliminate the extra impact of Endo H, the BLA was also expressed in Escherichia coli to get non-glycosylated Eco-BLA. A comparative study between non-glycosylated Eco-BLA and glycosylated Pic-BLA showed no obvious difference in thermostability. It is speculated that the glycosylation has little effect on the thermostability of α-amylase BLA.

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The purification and characterization of an extremely thermostable alpha-amylase from the hyperthermophilic archaebacterium Pyrococcus furiosus.

Cited by 154 publications

References 32 publications

Activity-Based Protein Profiling for the Identification of Novel Carbohydrate-Active Enzymes Involved in Xylan Degradation in the Hyperthermophilic Euryarchaeon Thermococcus sp. Strain 2319x1E

Activity-Based Protein Profiling for the Identification of Novel Carbohydrate-Active Enzymes Involved in Xylan Degradation in the Hyperthermophilic Euryarchaeon Thermococcus sp. Strain 2319x1E

Exploration for Thermostable β-Amylase of a Bacillus sp. Isolated from Compost Soil to Degrade Bacterial Biofilm

Expression of Bacillus licheniformis α-amylase in Pichia pastoris without antibiotics-resistant gene and effects of glycosylation on the enzymic thermostability

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