Tags

Type your tag names separated by a space and hit enter

Overexpression and characterization of dimeric and tetrameric forms of recombinant serine hydroxymethyltransferase from Bacillus stearothermophilus.
J Biosci. 2002 Jun; 27(3):233-42.JB

Abstract

Serine hydroxymethyltransferase (SHMT), a pyridoxal-5' -phosphate (PLP) dependent enzyme catalyzes the interconversion of L-Ser and Gly using tetrahydrofolate as a substrate. The gene encoding for SHMT was amplified by PCR from genomic DNA of Bacillus stearothermophilus and the PCR product was cloned and overexpressed in Escherichia coli. The purified recombinant enzyme was isolated as a mixture of dimer (90%) and tetramer (10%). This is the first report demonstrating the existence of SHMT as a dimer and tetramer in the same organism. The specific activities at 37 C of the dimeric and tetrameric forms were 6 7 U/mg and 4 1 U/mg, respectively. The purified dimer was extremely thermostable with a T(m) of 85 degrees C in the presence of PLP and L-Ser. The temperature optimum of the dimer was 80 degrees C with a specific activity of 32 4 U/mg at this temperature. The enzyme catalyzed tetrahydrofolate-independent reactions at a slower rate compared to the tetrahydrofolate-dependent retro-aldol cleavage of L-Ser. The interaction with substrates and their analogues indicated that the orientation of PLP ring of B. stearothermophilus SHMT was probably different from sheep liver cytosolic recombinant SHMT (scSHMT).

Authors+Show Affiliations

Department of Biochemistry, Indian Institute of Science, Bangalore 560 012, India.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12089472

Citation

Jala, Venkatakrishna R., et al. "Overexpression and Characterization of Dimeric and Tetrameric Forms of Recombinant Serine Hydroxymethyltransferase From Bacillus Stearothermophilus." Journal of Biosciences, vol. 27, no. 3, 2002, pp. 233-42.
Jala VR, Prakash V, Rao NA, et al. Overexpression and characterization of dimeric and tetrameric forms of recombinant serine hydroxymethyltransferase from Bacillus stearothermophilus. J Biosci. 2002;27(3):233-42.
Jala, V. R., Prakash, V., Rao, N. A., & Savithri, H. S. (2002). Overexpression and characterization of dimeric and tetrameric forms of recombinant serine hydroxymethyltransferase from Bacillus stearothermophilus. Journal of Biosciences, 27(3), 233-42.
Jala VR, et al. Overexpression and Characterization of Dimeric and Tetrameric Forms of Recombinant Serine Hydroxymethyltransferase From Bacillus Stearothermophilus. J Biosci. 2002;27(3):233-42. PubMed PMID: 12089472.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Overexpression and characterization of dimeric and tetrameric forms of recombinant serine hydroxymethyltransferase from Bacillus stearothermophilus. AU - Jala,Venkatakrishna R, AU - Prakash,V, AU - Rao,N Appaji, AU - Savithri,H S, PY - 2002/6/29/pubmed PY - 2002/12/5/medline PY - 2002/6/29/entrez SP - 233 EP - 42 JF - Journal of biosciences JO - J. Biosci. VL - 27 IS - 3 N2 - Serine hydroxymethyltransferase (SHMT), a pyridoxal-5' -phosphate (PLP) dependent enzyme catalyzes the interconversion of L-Ser and Gly using tetrahydrofolate as a substrate. The gene encoding for SHMT was amplified by PCR from genomic DNA of Bacillus stearothermophilus and the PCR product was cloned and overexpressed in Escherichia coli. The purified recombinant enzyme was isolated as a mixture of dimer (90%) and tetramer (10%). This is the first report demonstrating the existence of SHMT as a dimer and tetramer in the same organism. The specific activities at 37 C of the dimeric and tetrameric forms were 6 7 U/mg and 4 1 U/mg, respectively. The purified dimer was extremely thermostable with a T(m) of 85 degrees C in the presence of PLP and L-Ser. The temperature optimum of the dimer was 80 degrees C with a specific activity of 32 4 U/mg at this temperature. The enzyme catalyzed tetrahydrofolate-independent reactions at a slower rate compared to the tetrahydrofolate-dependent retro-aldol cleavage of L-Ser. The interaction with substrates and their analogues indicated that the orientation of PLP ring of B. stearothermophilus SHMT was probably different from sheep liver cytosolic recombinant SHMT (scSHMT). SN - 0250-5991 UR - https://www.unboundmedicine.com/medline/citation/12089472/Overexpression_and_characterization_of_dimeric_and_tetrameric_forms_of_recombinant_serine_hydroxymethyltransferase_from_Bacillus_stearothermophilus_ L2 - https://www.ias.ac.in/article/fulltext/jbsc/027/03/0233 DB - PRIME DP - Unbound Medicine ER -