Cloned (Comment) | Organism |
---|---|
two synthetic genes encoding the MTSase from Sulfolobus acidocaldarius strain ATCC 33909 are synthesized, recombinant expression of codon-optimized enzyme in Pichia pastoris strain KM71 (protein SP3.5) and in Escherichia coli strain BL21(DE3) (protein SEB24), method evaluation and optimization, overview | Sulfolobus acidocaldarius |
Protein Variants | Comment | Organism |
---|---|---|
additional information | comparative study of maltooligosyltrehalose synthase from Sulfolobus acidocaldarius expressed in Pichia pastoris strain KM71 and Escherichia coli strain BL21(DE3) for enzyme production. Comparison of the effect of different promoters on MTSase expression in Pichia pastoris, two different expression vectors are investigated: pPIC3.5 K, which contains an inducible alcohol oxidase (AOX1) promoter, and pGAPZA, which contains a constitutive glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter. The optimal pH, optimal temperature, pH stability, and temperature stability of MTSase expressed in different hosts are investigated and compared, overview. Both of the recombinant enzymes meet the demands of industrial trehalose production with respect to temperature stability | Sulfolobus acidocaldarius |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-[(1->4)-alpha-D-glucosyl]n-1-D-glucose | Sulfolobus acidocaldarius | - |
1-alpha-D-[(1->4)-alpha-D-glucosyl]n-1-alpha-D-glucopyranoside | - |
? | |
4-[(1->4)-alpha-D-glucosyl]n-1-D-glucose | Sulfolobus acidocaldarius ATCC 33909 / DSM 639 / JCM 8929 / NBRC 15157 / NCIMB 11770 | - |
1-alpha-D-[(1->4)-alpha-D-glucosyl]n-1-alpha-D-glucopyranoside | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Sulfolobus acidocaldarius | Q53688 | - |
- |
Sulfolobus acidocaldarius ATCC 33909 / DSM 639 / JCM 8929 / NBRC 15157 / NCIMB 11770 | Q53688 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme 4.6fold from Pichia pastoris strain KM71. and 5.6fold from Escherichia coli strain BL21(DE3), by ammonium sulfate fractionation, dialysis, and anion exchange chromatography | Sulfolobus acidocaldarius |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
maximum MTSase activity of crude enzyme expressed in Escherichia coli and Pichia pastoris strains reaches 31.6 and 133.0 U/ml, respectively | Sulfolobus acidocaldarius |
106.5 | - |
purified recombinant enzyme SP3.5, expressed in Pichia pastoris, pH 5.5, 60°C | Sulfolobus acidocaldarius |
116.5 | - |
purified recombinant enzyme, expressed in Escherichia coli, pH 5.5, 60°C | Sulfolobus acidocaldarius |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-[(1->4)-alpha-D-glucosyl]n-1-D-glucose | - |
Sulfolobus acidocaldarius | 1-alpha-D-[(1->4)-alpha-D-glucosyl]n-1-alpha-D-glucopyranoside | - |
? | |
4-[(1->4)-alpha-D-glucosyl]n-1-D-glucose | - |
Sulfolobus acidocaldarius ATCC 33909 / DSM 639 / JCM 8929 / NBRC 15157 / NCIMB 11770 | 1-alpha-D-[(1->4)-alpha-D-glucosyl]n-1-alpha-D-glucopyranoside | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 74000, recombinant enzyme, SDS-PAGE | Sulfolobus acidocaldarius |
Synonyms | Comment | Organism |
---|---|---|
maltooligosyltrehalose synthase | - |
Sulfolobus acidocaldarius |
MTSase | - |
Sulfolobus acidocaldarius |
SEB24 | - |
Sulfolobus acidocaldarius |
SP3.5 | - |
Sulfolobus acidocaldarius |
TreY | - |
Sulfolobus acidocaldarius |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
75 | - |
recombinant enzyme, SEB24 and SP3.5 | Sulfolobus acidocaldarius |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | 75 | temperature stability of recombinant SEB24 and SP3.5, both of them have a half-life of 12 h at 75°C. The half-lives of SEB24 and SP3.5 at 60°C are 6 d and 6.5 d, respectively. SP3.5 and SEB24 retain 80% and 97% of their original activity after incubation for 2 days at 60°C | Sulfolobus acidocaldarius |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.5 | - |
recombinant enzyme, SEB24 and SP3.5 | Sulfolobus acidocaldarius |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
both recombinant enzymes retain 80% relative activity after incubation at pH 4.0-7.5 for 24 h at 4°C. The pH stability of SEB24 over 24 h at pH 7.5-10.0 and 4°C is better than that of SP3.5, the relative activity of recombinant SEB24 is higher than that of recombinant SP3.5 from pH 3.5 to pH 5.5 | Sulfolobus acidocaldarius |