Kinetic resolution of N-protected amino acid esters in organic solvents catalyzed by a stable industrial alkaline protease

Chen, Shui‐Tein; Chen, Shiah-Yun; Hsiao, Shu-Chyong; Wang, Kung‐Tsung

doi:10.1007/bf01026757

Cited by 36 publications

(11 citation statements)

References 4 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Benzyl DL-alaninate was also enantioselectivity hydrolyzed. Alcalase was found to be very stable in organic solvents and useable as a catalyst for the resolution of N-benzyloxycarbonyl (Z) protected amino acid methyl esters (Chen et al, 1991). Hydrolyses were performed in t-butanol containing 5% (v/v) phosphate buffer (pH 8.2).…”

Section: A-chymotrypsin and Subtilisinmentioning

confidence: 99%

Enzymatic resolution of amino acids via ester hydrolysis

Miyazawa

1999

Amino Acids

View full text Add to dashboard Cite

show abstract

Section: A-chymotrypsin and Subtilisinmentioning

confidence: 99%

Enzymatic resolution of amino acids via ester hydrolysis

Miyazawa

1999

Amino Acids

View full text Add to dashboard Cite

show abstract

“…13 The alcalase had a half-life 12 h in t-butanol/ phosphate buffer (5%) and was stable in organic solvents containing aqueous solution (0-20%). Under those reaction conditions, the alcalase not only retained the esterification activity, but also had slightly improved catalytic efficiency.…”

Section: Resultsmentioning

confidence: 99%

Kinetic resolution of esters of amino acids in t‐butanol containing 5% water catalyzed by a stable industrial alkaline protease

1994

Self Cite

View full text Add to dashboard Cite

We developed a procedure for the resolution of esters of amino acids in 95% t-butanol, followed by saponification of the unreacted esters to afford both enantiomers with high yield and optical purity. The hydrolysis, catalyzed by alkaline protease, was conducted in a mixture of t-butanol(95%) and water (5%) at 25"C, with a pH controlled at pH 8.5 by the addition of NaOH (2 M). The hydrolyzed L-amino acid, which was insoluble under these conditions, precipitated during the course of hydrolysis. After separation of the precipitate, the pH of the filtrate was adjusted to 11.5 to sapomfy the unreacted ester. The D-antipode precipitated at pH 6.2-6.5. Both optically pure antipodes were obtained with high enantiomeric excesses and yields by simple filtration. o 1994 Wiey-Liss, Inc. KEY WORDS: resolution, D,L-&Oacid esters, alkaline protease, simple separation, high enantiomeric excess and yieldsThe search for proteases for peptide synthesis that are stable in organic solvents is the subject of extensive investigation. 's2 Several studies have demonstrated the possibility of using proteases to catalyze peptide synthesis in organic solvents, but a drawback of those reactions is the poor stability of the enzyme in organic solvents. Many immobilization processes have been developed to overcome this. Here we describe the use of an inexpensive enzyme, alcalase, as a catalyst for resolution of amino acids, some of which have uncommon side chains and are found in many biologically and pharmaceutically important peptides. 35 Most of these unnatural amino acids cannot be obtained by fermentation or recombinant DNA technology. Resolution is an effective way to produce optically pure unnatural amino acids.Alcalase is a proteolytic enzyme prepared from a selective strain of Bacillus lichenifomis. The major enzyme component in alcalase is subtilisin Carlsberg (alkaline protease A), which is a serine protease and which is widely used as additives in detergents as a digesting enzyme. This alcalase can maintain enzymatic activity and stability in organic solvents.6v7 Application of alcalase in organic synthesis has been limited, although it is a potent and inexpensive catalyst.'*' Scheme 1 shows the reaction sequence of the resolution. The hydrolysis catalyzed by alkaline protease was conducted in a mixture of t-butanol (95%) and water (5%) at 25°C with a pH controller at pH 8.5. The hydrolyzed amino acid was insoluble under these conditions and precipitated during the course of hydrolysis. The precipitate was separated by filtration, and the pH of the filtrate was adjusted to 11.5 to saponify the unreacted ester of the D-&O acid.

show abstract

“…The major component of this enzyme is subtilisin, which is a serine protease and is widely used as a detergent additive (Chen et al, 1991). The detergent industry has now emerged as the single major consumer of several other hydrolytic enzymes, including proteases acting in the alkaline pH range.…”

Section: Raimi Et Al 3189mentioning

confidence: 99%

“…An industrially important alkaline protease, isolated from a selective strain of Bacillus, has been shown to be useful as a catalyst for the resolution of N-protected amino acids having unusual side chains (Chen et al, 1991). The major component of this enzyme is subtilisin, which is a serine protease and is widely used as a detergent additive (Chen et al, 1991).…”

Section: Raimi Et Al 3189mentioning

confidence: 99%

Isolation and partial characterization of a protease enzyme from Thaumatococcus daniellii waste

Raimi¹,

Ojo²,

Elemo³

et al. 2011

Afr. J. Biotechnol.

View full text Add to dashboard Cite

A protease enzyme was isolated and partially purified from the pulp of Thaumatococcus daniellii fruit by gel filtration on sephadex G-75 followed by ion-exchange column chromatography on DEAEcellulose. The enzyme showed a specific activity of 4.75 × 10 -1 unit/mg protein and 6.93 × 10 -1 unit/mg protein, respectively after each purification procedure. The purified enzyme had a K m and V max of 2.0 × 10 -4 M and 1.53 mol/min, respectively, using casein as substrate. The enzyme had an optimum temperature of 35°C and functioned best at pH 7.0 with some residual activity at alkaline pH.

show abstract

Kinetic resolution of N-protected amino acid esters in organic solvents catalyzed by a stable industrial alkaline protease

Cited by 36 publications

References 4 publications

Enzymatic resolution of amino acids via ester hydrolysis

Enzymatic resolution of amino acids via ester hydrolysis

Kinetic resolution of esters of amino acids in t‐butanol containing 5% water catalyzed by a stable industrial alkaline protease

Isolation and partial characterization of a protease enzyme from Thaumatococcus daniellii waste

Contact Info

Product

Resources

About