Purification and properties of a novel enzyme, L‐α‐amino‐ϵ‐caprolactamase from <i>Cryptococcus laurentii</i>

Fukumura, Takashi; Talbot, Guylaine; Misono, Haruo; Teramura, Yasuo; Kato, Kazuo; Soda, Kenji

doi:10.1016/0014-5793(78)80240-3

Cited by 27 publications

(9 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…At first sight, one would expect a hydrolytic enzyme involved in lactam ring opening. The conversion of the related d , l -α-amino-ε-caprolactam by a combination of a racemase and a l -amino acid lactam hydrolase to yield l -lysine has been described (Payoungkiattikun et al 2017; Fukumura et al 1978; Ahmed et al 1982, 1986). Additionally, a putative hydrolase and aminotransferase have been reported for caprolactam metabolism.…”

Section: Introductionmentioning

confidence: 99%

Characterization of the caprolactam degradation pathway in Pseudomonas jessenii using mass spectrometry-based proteomics

Otzen

Palacio

Janssen

2018

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

Some bacterial cultures are capable of growth on caprolactam as sole carbon and nitrogen source, but the enzymes of the catabolic pathway have not been described. We isolated a caprolactam-degrading strain of Pseudomonas jessenii from soil and identified proteins and genes putatively involved in caprolactam metabolism using quantitative mass spectrometry-based proteomics. This led to the discovery of a caprolactamase and an aminotransferase that are involved in the initial steps of caprolactam conversion. Additionally, various proteins were identified that likely are involved in later steps of the pathway. The caprolactamase consists of two subunits and demonstrated high sequence identity to the 5-oxoprolinases. Escherichia coli cells expressing this caprolactamase did not convert 5-oxoproline but were able to hydrolyze caprolactam to form 6-aminocaproic acid in an ATP-dependent manner. Characterization of the aminotransferase revealed that the enzyme deaminates 6-aminocaproic acid to produce 6-oxohexanoate with pyruvate as amino acceptor. The amino acid sequence of the aminotransferase showed high similarity to subgroup II ω-aminotransferases of the PLP-fold type I proteins. Finally, analyses of the genome sequence revealed the presence of a caprolactam catabolism gene cluster comprising a set of genes involved in the conversion of caprolactam to adipate.Electronic supplementary materialThe online version of this article (10.1007/s00253-018-9073-7) contains supplementary material, which is available to authorized users.

show abstract

Section: Introductionmentioning

confidence: 99%

Characterization of the caprolactam degradation pathway in Pseudomonas jessenii using mass spectrometry-based proteomics

Otzen

Palacio

Janssen

2018

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

show abstract

“…Wehave purified both the enzymes to homogeneity and reported briefly their fundamental properties. 4…”

mentioning

confidence: 99%

Properties of .ALPHA.-amino-.EPSILON.-caprolactam racemase from Achromobacter obae.

Ahmed

Esaki

Tanaka

et al. 1983

Agricultural and Biological Chemistry

View full text Add to dashboard Cite

a-Amino-e-caprolactam racemase, which occurs in the cytoplasmic fraction of Achromobacter obae, has been purified to homogeneity. It has a monomericstructure with a molecular weight of approximately 50,000. The absorption spectrum of the enzymeexhibits maximaat 280 and 412 nm at pH 7.3, and is independent of pH from 6.0 to 8.0. One mole ofpyridoxal 5'-phosphate is bound per mol of the enzyme.Incubation of the enzymewith hydroxylamineresulted in the formation of the apoenzyme. d-and L-a-Amino-a-caprolactamsare the only substrates. The maximum activity is found at pH 8.8 for both the isomers. Michaelis constants are as follows: 8him for D-a-amino-ecaprolactam, 6 mMfor L-a-amino-e-caprolactam and 2. 1 x 10~7 m for pyridoxal 5'-phosphate. The enzyme is inhibited significantly by CuSO4, HgCl2, thiol reagents such as N-ethylmaleimide and /?-chloromercuribenzoate, and carbonyl reagents (e.g., phenylhydrazine and hydroxylamine). aAmino-e-caprolactam racemase catalyzes the a-proton exchange of the substrate with deuteron during racemization in deuterium oxide. Various racemases have been demonstrated in bacteria and actinomycetes, but only a few enzymes have been purified to homogeneity for enzymological studies: pyridoxal 5'-phosphate dependent amino acid racemases,6~8) imino acid racemases with no cafactor requirement,9'10) an ATP dependent phenylalanine racemase11] and a Mg++ dependent non-amino acid racemase, mandelate racemase.12) aAmino-e-caprolactam racemase is unique amongracemases in that it acts exclusively on a synthetic non-carboxylic compound i.e., an intramolecular cyclic amide of L-lysine.5) We have shown that this enzyme requires pyridoxal S'-phosphate as a coenzyme.5) Thus, the enzyme is the only pyridoxal 5'-phosphate enzyme acting on an a-carboxyamide derivative of an amino acid. Wehere describe the localization of the enzyme in bacterial cells and the physicochemical and enzymological properties of the enzyme purified from Achromobacter obae.

show abstract

“…The process is composed of two new enzymatic reactions: the stereo-selective hydrolysis of L-a-amino-c:-caprolactam to L-Iysine by L-aamino-c:-caprolactamase (EC class 3.5.2) of an yeast, and the racemization of a-amino-c:-caprolactam by bacterial a-amino-c:-caprolactam racemase (EC class 5.1.1 ).1,2) Thus, L-Iysine is produced with a yield of nearly 100%.3) We have purified both the enzymes to homogeneity and reported briefly their fundamental properties. 4,5) Various racemases have been demonstrated in bacteria and· actinomycetes, but only a few enzymes have been purified to homogeneity for enzymological studies: pyridoxal 5I -phosphate dependent amino acid racemases,6"-'8) imino acid racemases with no cafactor requirement,9,10) an ATP dependent phenylalanine racemase 11 ) and a Mg+ + dependent non-amino acid racemase, mandelate racemase. 12) aAmino-c:-caprolactam racemase is unique among racemases in that it acts exclusively on a synthetic non-carboxylic compound i.e., an intramolecular cyclic amide of L-lysine.…”

mentioning

confidence: 99%

Properties of α-Amino-ε-caprolactam Racemase fromAchromobacter obae

Ahmed¹,

Esaki²,

Tanaka³

et al. 1983

Agricultural and Biological Chemistry

View full text Add to dashboard Cite

Purification and properties of a novel enzyme, L‐α‐amino‐ϵ‐caprolactamase from Cryptococcus laurentii

Cited by 27 publications

References 17 publications

Characterization of the caprolactam degradation pathway in Pseudomonas jessenii using mass spectrometry-based proteomics

Characterization of the caprolactam degradation pathway in Pseudomonas jessenii using mass spectrometry-based proteomics

Properties of .ALPHA.-amino-.EPSILON.-caprolactam racemase from Achromobacter obae.

Properties of α-Amino-ε-caprolactam Racemase fromAchromobacter obae

Contact Info

Product

Resources

About