This summer, we have characterized 105 BioBricks which could either be used directly or serve as a universal tool readily for potential scientific or engineering use.

Those Biobricks could be divided into 4 groups.

n-Butanol Production

BBa_K1462060: adhE2, the gene encoding the NADH-dependent aldehyde/alcohol dehydrogenase responsible for butanol production in alcohologenic cultures of Clostridium acetobutylicum. It participates in generation of useful aldehyde, ketone, or alcohol groups during biosynthesis of various metabolites. And sometimes alcohol dehydrogenases catalyze the opposite reaction as part of fermentation to ensure a constant supply of NAD+.

BBa_K1462000 to BBa_K1462080 are basic parts which are basic components of the pathway, including some enzymes and terminators.
BBa_K1462110 to BBa_K1462200 are proteins fused with GFP used to verify the location of enzymes, in cytoplasm or in mitochondrial matrix.
BBa_K1462210 to BBa_K1462210 are responsible for the construction of n-butanol biosynthesis pathway in cytoplasm or in mitochondrial matrix.

CO2 Fixation

BBa_K1462380: CA (carbonic anhydrases or carbonate dehydratases), form a family of enzymes that catalyze the rapid interconversion of carbon dioxide and water to bicarbonate and protons (or vice versa), a reversible reaction that occurs relatively slowly in the absence of a catalyst. The reaction catalyzed by carbonic anhydrase is: H2CO3↔CO2+ H2O

BBa_K1462310 to BBa_K1462450 are BioBricks of CO2 fixation pathway, which utilize the high-concentration ATP and CO2 around mitochondria to efficiently fix CO2 and improve the C sequestration of yeast. This pathway increases the yield of pyruvate for butanol production as the standing point. BBa_K1462980 and BBa_K1462990 are EroGL/EroGS, two E.coli protein-folding chaperons, which stimulate functional expression of RuBisCo.

Scaffold Proteins

BBa_K1462460, BBa_K1462470 and BBa_K1462480 are corresponding to the design of GBD, SH3, PDZ ligands; BBa_K1462501, while BBa_K1462502, BBa_K1462503 are relevant to the construction of GBD, SH3, PDZ domains with their linkers. BBa_K1462690 to BBa_K1462840 are different proportions of scaffold proteins, GBD(x), SH3(y) and PDZ(z). Our team concentrates on constructing a tool with a wide range of applications which would beneficial to the researchers from all over the world. With the utilization of the tools, you could control the direction of metabolic flux and enhance the efficiency of biochemical reactions via regulating the ratios of the enzymes and restricting the reactions in a fixed domain. Meanwhile, it achieves the quantification of the projects by altering the number of each enzyme and obtaining the optimum ratio.

Leading Peptides

BBa_K1462850 to BBa_K1462970 are leading peptides in yeast. We chose 6 locations in yeast, Plasma Membrane, Vacuolar membrane, Nucleus, Peroxisome, Cytoderm and Endoplasmic reticulum. The verifications of these leading peptides are the first step for us to take further studies in subcellular compartments.

SCUT 2014 iGEM team Parts

<groupparts>iGEM014 SCUT</groupparts>