Papain-catalysed synthesis of dipeptides: A novel approach using free amino acids as nucleophiles

Stehle, Peter; Bahsitta, Hans-Peter; Monter, B.; Fürst, Peter

doi:10.1016/0141-0229(90)90181-o

Cited by 34 publications

(8 citation statements)

References 18 publications

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“…, 2007). Enzyme‐catalyzed reactions give access to a large number of peptides synthesized by different proteases under various conditions (Stehle et al. , 1990; Bordusa, 2002; Yokozeki and Hara, 2005; Li and Kanerva, 2006; Heck et al.…”

Section: Introductionmentioning

confidence: 99%

Simple enzymatic procedure for l‐carnosine synthesis: whole‐cell biocatalysis and efficient biocatalyst recycling

Heyland

Antweiler

Lutz

et al. 2009

Microbial Biotechnology

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Summaryβ‐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.

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Section: Introductionmentioning

confidence: 99%

Simple enzymatic procedure for l‐carnosine synthesis: whole‐cell biocatalysis and efficient biocatalyst recycling

Heyland

Antweiler

Lutz

et al. 2009

Microbial Biotechnology

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“…Enzymatic Peptide Synthesis -Enzymatic synthesis of Z-Phe-Arg-SerNH 2 using Z-Phe-ArgOMe HCl (ester) and Ser-NH 2 HCl (amine) was performed based on methods described by Stehle et al [10] and Lozano et al [11]. rFheCL1 was activated to a final concentration of 0.05mg/ml.…”

Section: Methodsmentioning

confidence: 99%

Peptide synthesis by recombinant Fasciola hepatica cathepsin L1

2006

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Synthesis of the tripeptide Z-Phe-Arg-SerNH 2 has been accomplished by a recombinant cysteine protease, cathepsin L1 from liver fluke (Fasciola hepatica), using Z-Phe-Arg-OMe as acyl acceptor and SerNH 2 as nucleophile in 0.1 M ammonium acetate pH 9.0-12.5% v/v acetonitrile at 37 o C. LC-MS detection indicated tripeptide formation after 10 min, continuing up to 5.5 h. The ester Z-Phe-Arg-OMe was detected throughout the experiment but the hydrolysis product Z-Phe-Arg-OH appeared early and in quite large amounts. We believe that this is the first application of a parasite protease in enzymatic peptide synthesis.Keywords: Fasciola hepatica cathepsin L1, recombinant, enzymatic peptide synthesis, cysteine proteinase 2 Abbreviations: LC-MS, liquid chromatography-mass spectrometry; m/z, mass-to-charge ratio; rFheCL1, recombinant Fasciola hepatica cathepsin L1; Z, benzoyloxycarbonyl.

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“…Recently we have been engaged in studies dealing with the development of novel synthesis procedures using native and/or immobilized plant proteases as biocatalysts [15,16]. Compared with classical synthesis methods, this biotechnological approach offers advantages, like (stereo-)selectivity of the reaction, minimal protection of functional groups and simplified purification procedures.…”

Section: Synthetic Short-chain Peptides: Propertiesmentioning

confidence: 99%