Difference between revisions of "Part:BBa K633003"

Revision as of 01:47, 18 October 2011

Extracellular Succrase Sacc

Sacc Extracellular Sucrase

This extracellular sucrase is an enzyme from Zymomonas mobilis. The expression product is different from SacB; showing sucrase but not levansucrase activity. In E. coli BL21, after induction of the T7 polymerase expression system with IPTG. Subcellular fractionation of the E. coli clone carrying plasmid pLSS2811 showed that more than 70% of the sucrase activity could be detected in the cytoplasmic fraction, suggesting that the enzyme was soluble and not secreted in E. coli. Its structure is monomeric, with an optimum temperature 20 to 40 ° C, pH of 2.5-7.5 and a size of 48 kDa. It also has a free N-terminus.

Invertase hydrolyses the non-reducing β-d-fructofuranoside residues of sucrose to yield invert sugar. The invert sugar released is then reacted with 3.5 dinitrosalicylic acid (DNS). The colour change produced is proportional to the amount of invert sugar released, which in turn is proportional to the invertase activity present in the sample. The absorbance is measured at 540 nm and converted into micromoles of reducing sugar produced using a calibration curve. One invertase unit is the amount of enzyme which will produce 1 micromole of reducing sugar (expressed as invert sugar) per minute under the conditions specified in this procedure.

This part was cloned from Zymmomonas mobilis in the FEMSA Biotechnology Center by phD. Jose Manuel Aguilar, in team TEC-Monterrey's campus.

The cloning technique was the next:

Kannan R, Mukundan G, Aït-Abdelkader N, Augier-Magro V, Baratti J, Gunasekaran P. Molecular cloning and characterization of the extracellular sucrase gene (sacC) of Zymomonas mobilis. Arch Microbiol. 1995 Mar;163(3):195-204.

Sequence and Features