Identification, Cloning, and Sequencing of the Genes Involved in the Conversion of<scp>D</scp>,<scp>L</scp>-2-Amino-<i>Δ</i><sup>2</sup>-Thiazoline-4-Carboxylic Acid to …

Ohmachi, Tetsuo; Nishino, Mizuka; Kawata, Maki; Edo, Namiko; Funaki, Hiroko; Narita, Megumi; Mori, Kazuyuki; Tamura, Yoshiharu; Asada, Yoshihiro

doi:10.1271/bbb.66.1097

Cited by 20 publications

(8 citation statements)

References 24 publications

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“…ATC-utilising enzymes are discussed to be important in amino acid synthesis in bacteria but could as well be involved in the synthesis or degradation of thiazolin-containing molecules such as certain bacteriocins or siderophores (Shiba et al, 2002). Bacteria expressing atc gene products have become important for commercial production of l-cysteine (Ohmachi et al, 2002). The examples described above demonstrate that the S. meliloti accessory gene pool also includes modules that most probably originate from nonrhizobial species.…”

Section: Degradative Capabilities Encoded On Psmesm11cmentioning

confidence: 99%

The complete genome sequence of the dominant Sinorhizobium meliloti field isolate SM11 extends the S. meliloti pan-genome

Schneiker-Bekel

Wibberg

Bekel

et al. 2011

Journal of Biotechnology

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Section: Degradative Capabilities Encoded On Psmesm11cmentioning

confidence: 99%

The complete genome sequence of the dominant Sinorhizobium meliloti field isolate SM11 extends the S. meliloti pan-genome

Schneiker-Bekel

Wibberg

Bekel

et al. 2011

Journal of Biotechnology

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“…Several genes involved in the L-NCC pathway have been cloned from Pseudomonas sp. Strain ON-4a (Ohmachi et al, 2002) and strain BS (Shiba et al, 2002). However, the CD gene involved in this process, which is responsible for the decomposition of L-cysteine, is poorly understood and validated.…”

Section: Discussionmentioning

confidence: 99%

Cloning, expression and characterization of L-cysteine desulfhydrase gene from Pseudomonas sp. TS1138

Bai

Yang

et al. 2007

Front. Biol. China

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L-cysteine desulfhydrase (CD) plays an important role in L-cysteine decomposition. To identify the CD gene in Pseudomonas sp. TS1138 and investigate its effect on the L-cysteine biosynthetic pathway, the CD gene was cloned from Pseudomonas sp. TS1138 by polymerase chain reaction (PCR) method. The nucleotide sequence of CD gene was determined to be 1,215 bp, and its homology with other sequences encoding CD was analyzed. Then the CD gene was subcloned into pET-21a(+) vector and expressed in Escherichia coli (E. coli) by isopropyl-b-D-thiogalactopyranoside (IPTG) inducement. The recombinant CD was purified by Ni-NTA His-Bind resin, and its activity was identified by the CD activity staining. The enzymatic properties of the recombinant CD were characterized and its critical role involved in the L-cysteine biosynthetic pathway was also discussed.

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“…An electrophoresis calibration kit (Pharmacia Biotec) was used for the estimation of the molecular mass as standard proteins. Protein sequencing of the purified r-thiolase was carried out on a protein sequencer (Model PPSQ-10, SHIMADZU, Japan) according to the method described previously 17.…”

Section: Methodsmentioning

confidence: 99%

Expression, Identification and Purification of Dictyostelium Acetoacetyl-CoA Thiolase Expressed in Escherichia coli

Tanaka¹,

Shima²,

Ogawa³

et al. 2011

Int. J. Biol. Sci.

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Acetoacetyl-CoA thiolase (AT) is an enzyme that catalyses the CoA-dependent thiolytic cleavage of acetoacetyl-CoA to yield 2 molecules of acetyl-CoA, or the reverse condensation reaction. A full-length cDNA clone pBSGT-3, which has homology to known thiolases, was isolated from Dictyostelium cDNA library. Expression of the protein encoded in pBSGT-3 in Escherichia coli, its thiolase enzyme activity, and the amino acid sequence homology search revealed that pBSGT-3 encodes an AT. The recombinant AT (r-thiolase) was expressed in an active form in an E. coli expression system, and purified to homogeneity by selective ammonium sulfate fractionation and two steps of column chromatography. The purified enzyme exhibited a specific activity of 4.70 mU/mg protein. Its N-terminal sequence was (NH 2 )-Arg-Met-Tyr-Thr-Thr-Ala-Lys-Asn-Leu-Glu-, which corresponds to the sequence from positions 15 to 24 of the amino acid sequence deduced from pBSGT-3 clone. The r-thiolase in the inclusion body expressed highly in E. coli was the precursor form, which is slightly larger than the purified r-thiolase. When incubated with the cell-free extract of Dictyostelium cells, the precursor was converted to the same size to the purified r-thiolase, suggesting that the presequence at the N-terminus is removed by a Dictyostelium processing peptidase.

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Identification, Cloning, and Sequencing of the Genes Involved in the Conversion ofD,L-2-Amino-Δ²-Thiazoline-4-Carboxylic Acid to …

Cited by 20 publications

References 24 publications

The complete genome sequence of the dominant Sinorhizobium meliloti field isolate SM11 extends the S. meliloti pan-genome

The complete genome sequence of the dominant Sinorhizobium meliloti field isolate SM11 extends the S. meliloti pan-genome

Cloning, expression and characterization of L-cysteine desulfhydrase gene from Pseudomonas sp. TS1138

Expression, Identification and Purification of Dictyostelium Acetoacetyl-CoA Thiolase Expressed in Escherichia coli

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Identification, Cloning, and Sequencing of the Genes Involved in the Conversion ofD,L-2-Amino-Δ2-Thiazoline-4-Carboxylic Acid to …

Cited by 20 publications

References 24 publications

The complete genome sequence of the dominant Sinorhizobium meliloti field isolate SM11 extends the S. meliloti pan-genome

The complete genome sequence of the dominant Sinorhizobium meliloti field isolate SM11 extends the S. meliloti pan-genome

Cloning, expression and characterization of L-cysteine desulfhydrase gene from Pseudomonas sp. TS1138

Expression, Identification and Purification of Dictyostelium Acetoacetyl-CoA Thiolase Expressed in Escherichia coli

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Identification, Cloning, and Sequencing of the Genes Involved in the Conversion ofD,L-2-Amino-Δ²-Thiazoline-4-Carboxylic Acid to …