Biosynthesis of peptidoglycan in Gaffkya homari. The incorporation of peptidoglycan into the cell wall and the direction of transpeptidation.

Wall membrane enzyme preparations from Gaffkya homari catalyze the formation of peptidoglycan from the precursor pairs: UDP-N-acetylglucosamine + UDP-N-acetylmuramyl-pentapeptide (UDP-MurNAc-Ala-DGlu-Lys-DAla-DAla) and also from UDP-N-acetylglucosamine + UDP-N-acetylmuramyl-tetrapeptide (UDP-MurNAc-Ala-DGlu-Lys-DAla). Part of the reaction products is soluble in 2% sodium dodecylsfulfate whereas the other part is bound to pre-existing cell wall peptidoglycan. The incorporation into cell wall takes place by a transpeptidation reaction in which the D-alanyl-D-alanine sequences in the pre-existing cell wall function as donors and the epsilon-amino groups of the lysine residues in the newly synthesized peptidoglycan strands function as acceptors. Nepsilon-D-Alanyl-lysine linkages are formed. At saturating concentration of UDP-N-acetylglucosamine, the enzyme system exhibits similar apparent Km values (30--80 muM) for UDP-MurNAc-pentapeptide and UDP-MurNAc-tetrapeptide both for the formation of cell-wall bound peptidoglycan and total (i.e. soluble + cell-wall-bound) peptidoglycan. The V values are also in the same order of magnitude (270-650 pmol x min-1 x mg of protein -1). However, UDP-MurNAc-tetrapeptide was a slightly better substrate than UDP-MurNAc-pentapeptide for the formation of cell-wall-bound peptidoglucan. The synthesis of total and cell-wall-bound peptidoglycan from UDP-MurNAc-pentapeptide was competitively inhibited by UDP-MurNAc-tetrapeptide and vice versa. UDP-MurNAc-tripeptide and both UDP-Mur-NAc-pentapeptide and UDP-Mur-NAc-tetrapeptide in which the epsilon-amino group of the lysine residue was substituted by an acetyl group were utilized less efficiently than UDP-MurNAc-pentapeptide and UDP-MurNAc-tetrapeptide for the formation of soluble peptidoglycan; they were exceedingly poor substrates for the formation of cell-wall-bound peptidoglycan.