Engineering a Function into a Glycosyltransferase

Abstract

As glycosyltransferases found in nature often show distinct substrate specificity, glycosyltransferase engineering is an important research field. In this work, we were able to introduce an activity into a glycosyltransferase involved in natural product (landomycin E) biosynthesis. This was achieved by recognizing hot spot amino acids in glycosyltransferases which are strongly involved in determining substrate specificity.

Cloning and Sequencing of the Biosynthetic Gene Cluster for Saquayamycin Z and Galtamycin B and the Elucidation of the Assembly of Their Saccharide Chains

“…SaqJ (15) and SaqJ1 (22) resemble transport proteins that are likely responsible for transport of the secondary metabolites across the membrane.…”

“…Further studies on what controls the level of iteration will be performed, similar to the studies carried out with glycosyltransferases involved in landomycin biosynthesis. [22] Interestingly, the mono-glycosylated galtamycinone instead of the expected fully glycosylated galtamycin B accumulated in the saqGT2 and saqGT3 mutants. This suggests that glycosyltransferases that act on the tetrasaccharide chain are only acting on the angucycline and not on the tetracenequinone.…”

Enzymatic Glycosylation of Nonbenzoquinone Geldanamycin Analogs via Bacillus UDP-Glycosyltransferase

“…The substrate flexibility of bacterial glycosyltransferases offers the possibility of producing antibiotics or other natural compounds with drastically changed glycosylation patterns (17,32). Therefore, the exploitation of the substrate flexibility becomes an important focus in biosynthetic approaches to the chemical diversity of medically active compounds.…”

“…These sugars are often essential for the pharmacological properties, especially the water solubility and/or the biological activity, of the compounds. Several complementary strategies, including semisynthesis, pathway engineering, and in vitro enzymatic glycosylation techniques, have emerged from recent studies as effective means of altering the natural product sugar structures (9,17,31,32). The clinical evaluation of GM and some GM derivatives has not been pursued due to their severe toxicity and poor water solubility (22).…”

“…Several complementary strategies, including semisynthesis, pathway engineering, and in vitro enzymatic glycosylation techniques, have emerged from recent research as effective means of altering the natural product sugar structures (6,8,9,11,17,32). A well-known example of the improvement in water solubility is the discovery of naturally occurring glycosides such as 7␤-xylosyl-10 deacetyl paclitaxel, a taxane glycocojugate (20).…”