xylA cloning and sequencing and biochemical characterization of xylose isomerase from Thermotoga neapolitana

Vieille, Claire; Hess, J. Michael; Kelly, Robert M.; Zeikus, J. G.

doi:10.1128/aem.61.5.1867-1875.1995

Cited by 105 publications

(73 citation statements)

References 36 publications

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“…The retention of high percentage of TnapXI activity at slightly acidic to neutral pH is very useful in industries for HFCS production at elevated temperature, because at high temperatures and alkaline pH browning by-products are formed by non-enzymatic reactions between reducing sugars and ɛ-amino group of lysine (Lys) residues. This Maillard reaction also caused inactivation of the enzyme [38]. Similarly, Vieille et al [38] also reported that TNXI retained 100% activity at pH range of 6.8-7.3 for at least 30 min when incubated at 90°C.…”

Section: Discussionmentioning

confidence: 86%

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Cloning, purification, and characterization of xylose isomerase from Thermotoga naphthophila RKU‐10

Fatima

Aftab

Haq

2016

J. Basic Microbiol.

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A 1.3 kb xyl-A gene encoding xylose isomerase from a hyperthermophilic eubacterium Thermotoga naphthophila RKU-10 (TnapXI) was cloned and over-expressed in Escherichia coli to produce the enzyme in mesophilic conditions that work at high temperature. The enzyme was concentrated by lyophilization and purified by heat treatment, fractional precipitation, and UNOsphere Q anion-exchange column chromatography to homogeneity level. The apparent molecular mass was estimated by SDS-PAGE to be 49.5 kDa. The active enzyme showed a clear zone on Native-PAGE when stained with 2, 3, 5-triphenyltetrazolium chloride. The optimum temperature and pH for D-glucose to D-fructose isomerization were 98 °C and 7.0, respectively. Xylose isomerase retains 85% of its activity at 50 °C (t1/2 1732 min) for 4 h and 32.5% at 90 °C (t1/2 58 min) for 2 h. It retains 90-95% of its activity at pH 6.5-7.5 for 30 min. The enzyme was highly activated (350%) with the addition of 0.5 mM Co(2+) and to a lesser extent about 180 and 80% with the addition of 5 and 10 mM Mn(2+) and Mg(2+) , respectively but it was inhibited (54-90%) in the presence of 0.5-10 mM Ca(2+) with respect to apo-enzyme. D-glucose isomerization product was also analyzed by Thin Layer Chromatography (Rf 0.65). The enzyme was very stable at neutral pH and sufficiently high temperature and required only a trace amount of Co(2+) for its optimal activity and stability. Overall, 52.2% conversion of D-glucose to D-fructose was achieved by TnapXI. Thus, it has a great potential for industrial applications.

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Section: Discussionmentioning

confidence: 86%

“…This Maillard reaction also caused inactivation of the enzyme [38]. Similarly, Vieille et al [38] also reported that TNXI retained 100% activity at pH range of 6.8-7.3 for at least 30 min when incubated at 90°C. However, F. gondwanense XI retained 40% of maximum activity at pH 5.0-8.0, 4°C [1].…”

Section: Discussionmentioning

confidence: 86%

Cloning, purification, and characterization of xylose isomerase from Thermotoga naphthophila RKU‐10

Fatima

Aftab

Haq

2016

J. Basic Microbiol.

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“…Most of the known ADHs from thermophiles are NAD(P)-dependent, with the exception of the Methanoculleus thermophilicus ADH that uses cofactor F 420 in the place of NAD(P) ( Table 1). This similarity of cofactor utilisation with their mesophilic counterparts indicates that they share a common catalytic mechanism [20,21].…”

Section: Adhs From Thermophilic Bacteria and Archaeamentioning

confidence: 83%

Alcohol dehydrogenases from thermophilic and hyperthermophilic archaea and bacteria

2003

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Many studies have been undertaken to characterise alcohol dehydrogenases (ADHs) from thermophiles and hyperthermophiles, mainly to better understand their activities and thermostability. To date, there are 20 thermophilic archaeal and 17 thermophilic bacterial strains known to have ADHs or similar enzymes, including the hypothetical proteins. Some of these thermophiles are found to have multiple ADHs, sometimes of different types. A rigid delineation of amino acid sequences amongst currently elucidated thermophilic ADHs and similar proteins is phylogenetically apparent. All are NAD(P)-dependent, with one exception that utilises the cofactor F 420 instead. Within the NAD(P)-dependent group, the thermophilic ADHs are orderly clustered as zinc-dependent ADHs, short-chain ADHs, and ironcontaining/activated ADHs. Distance matrix calculations reveal that thermophilic ADHs within one type are homologous, with those derived from a single genus often showing high similarities. Elucidation of the enzyme activity and stability, coupled with structure analysis, provides excellent information to explain the relationship between them, and thermophilic ADHs diversity.

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“…Comparison of the barley protein to the corresponding prokaryotic proteins. The best known xylose isomerases have been divided in two families (Vieille et al, 1995) or into two clusters within the same family (Meaden et al, 1994). The barley isomerase has an insertion of about 40 amino acids at its amino terminus when compared to the family-I1 proteins, which themselves are about 50 residues longer than the family-I proteins (Table 2).…”

Section: Effect Of Phmentioning

confidence: 99%

Protein Purification, and Cloning and Characterization of the cDNA and Gene for Xylose Isomerase of Barley

Kristo¹,

Saarelainen²,

Fagerstrom³

et al. 1996

European Journal of Biochemistry

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The first eukaryotic xylose isomerase protein was purified from barley Hordeum vulgare. The enzyme requires Mnz+ for its activity and is fairly thermostable, with the optimum temperature being 60°C. It showed maximum activity over a broad pH range (7.0-9.0). The molecular mass of the monomer was about 50 000 Da based on the SDSPAGE, and the calculated value from the cDNA-deduced polypeptide sequence was 53620 Da. A relative mass estimation of 100000 Da was obtained from the Superose 12 chromatography, suggesting that the barley enzyme is a dimer. The cloned corresponding cDNA sequence of 1710 nucleotides encoded a polypeptide of 480 amino acids. The genomic sequence of 4473 nucleotides, revealed that the isomerase gene contained 20 introns, all starting with GT and ending with AG. One large intron was located in the S'untranslated region. The barley isomerase has an insertion of about 40 residues at its amino terminus when compared to the prokaryotic cluster (family) I1 isomerases; cluster (family) I and cluster (family) I1 isomerases vary from the former in an insertion of around 50 residues at their amino termini. Comparison of the barley protein with the prokaryotic isomerases shows that the conserved catalytic and metal binding regions are also well conserved in barley.

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xylA cloning and sequencing and biochemical characterization of xylose isomerase from Thermotoga neapolitana

Cited by 105 publications

References 36 publications

Cloning, purification, and characterization of xylose isomerase from Thermotoga naphthophila RKU‐10

Cloning, purification, and characterization of xylose isomerase from Thermotoga naphthophila RKU‐10

Alcohol dehydrogenases from thermophilic and hyperthermophilic archaea and bacteria

Protein Purification, and Cloning and Characterization of the cDNA and Gene for Xylose Isomerase of Barley

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