Simple enzymatic procedure for L-carnosine synthesis: whole-cell biocatalysis and efficient biocatalyst recycling.
β-Peptides and their derivates are usually stable to proteolysis and have an increased half-life compared with α-peptides. Recently, β-aminopeptidases were described as a new enzyme class that enabled the enzymatic degradation and formation of β-peptides. As an alternative to the existing chemical synthesis routes, the aim of the present work was to develop a whole-cell biocatalyst for the synthesis and production of β-peptides using this enzymatic activity. For the optimization of the reaction system we chose the commercially relevant β,α-dipeptide L-carnosine (β-alanine-L-histidine) as model product. We were able to show that different recombinant yeast and bacteria strains, which overexpress a β-peptidase, could be used directly as whole-cell biocatalysts for the synthesis of L-carnosine. By optimizing relevant reaction conditions for the best-performing recombinant Escherichia coli strain, such as pH and substrate concentrations, we obtained high l-carnosine yields of up to 71%. Long-time as well as biocatalyst recycling experiments indicated a high stability of the developed biocatalyst for at least five repeated batches. Application of the recombinant E. coli in a fed-batch process enabled the accumulation of l-carnosine to a concentration of 3.7 g l(-1).
Laboratory of Chemical Biotechnology, TU Dortmund, 44221 Dortmund, Germany.
Jan
J
Heyland
Nicolai
N
Antweiler
Jochen
J
Lutz
Tobias
T
Heck
Birgit
B
Geueke
Hans-Peter E
HP
Kohler
Lars M
LM
Blank
Andreas
A
Schmid
D001419:Bacteria / Q000201:enzymology / Q000378:metabolism*; D055162:Biocatalysis; D001709:Biotechnology / Q000379:methods*; D002336:Carnosine / Q000096:biosynthesis*; D004798:Enzymes / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015003:Yeasts / Q000201:enzymology / Q000378:metabolism*
D004798:Enzymes; D011994:Recombinant Proteins; D002336:Carnosine; D010447:Peptide Hydrolases
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