Purification, characterization, and genetic analysis of a leucine aminopeptidase from Aspergillus sojae

Chien, Hungchien Roger; Lin, Long-Liu; Chao, Shiou-Huei; Chen, Chun-Chang; Wang, Wen-Ching; Shaw, Chin-Ying; Tsai, Ying-Chie; Hu, Hui-Yu; Hsu, Wen-Hwei

doi:10.1016/s0167-4781(02)00307-x

Cited by 63 publications

(39 citation statements)

References 41 publications

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“…Most fungal phytases characterized till date exhibit temperature optima in the range of 50-60°C. Extracellular enzymes produced by mesophilic fungi generally display temperature optima below 60°C although a few of them display higher temperature optima (Manzanares et al 1997;Chien et al 2002). This is also evident in the present study.…”

Section: Resultssupporting

confidence: 83%

Extracellular phytase from Aspergillus niger CFR 335: purification and characterization

Gunashree

Venkateswaran

2014

J Food Sci Technol

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Phytase, that is extensively used as a feed additive is capable of hydrolyzing phytic acid, an antinutrient found in about 60-80 % of all the plant commodities. This enzyme improves the bioavailability of essential minerals such as Ca 2+ , Mg 2+ , P, Zn 2+ , Fe 3+ , that are bound to phytic acid. An extracellular phytase from a local fungal isolate, Aspergillus niger CFR 335 was purified to homogeneity through a threestep column chromatography using DEAE-Sephadex anion exchanger. An active fraction of the enzyme was obtained with NaCl gradient of 2.5 M in DEAE Sephadex column. The enzyme was purified up to 16 fold with a yield of 28.5 %. Substrate specificity studies revealed a highest specific activity of 32.6±3.1 U/mg for sodium phytate with the K m value of 0.08±0.1 mM. The molecular weight of the enzyme was 66 kDa with an optimum temperature of 30°C and pH 4.5. Up to 80 % of the activity was retained even after storing the enzyme for 6 months at 4°C.

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Section: Resultssupporting

confidence: 83%

Extracellular phytase from Aspergillus niger CFR 335: purification and characterization

Gunashree

Venkateswaran

2014

J Food Sci Technol

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“…3, Table 6). ruLap1, fulap1 and A. oryzae 31 kDa Lap (Chien et al, 2002) structurally belong to the same subfamily M28E as Vibrio and Aeromonas leucyl aminopeptidases (Toma & Honma, 1996) (MEROPS>M28?002 and MEROPS>M28?004, respectively) (Fig. 4, Table 7).…”

Section: Discussionmentioning

confidence: 96%

“…A 1200 bp fragment containing the nucleotide sequence of the gene encoding an Aspergillus oryzae/Aspergillus sojae 31 kDa Lap (orLap1) (Nakadai et al, 1973;Chien et al, 2002;US Patent no. 5,994,113) was obtained by PCR of A. oryzae genomic DNA using the oligonucleotides 59-ATGCGTTTCCTCCCCTGCATCGCG-39 (sense) and 59-CAGCGAATCTGCGAAGGCAAGCTC-39 (antisense).…”

Section: Cloning Of Genes Encoding T Rubrum and A Fumigatus Lapsmentioning

confidence: 99%

Aminopeptidases and dipeptidyl-peptidases secreted by the dermatophyte Trichophyton rubrum

et al. 2005

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The nature of secreted aminopeptidases in Trichophyton rubrum was investigated by using a reverse genetic approach. T. rubrum genomic and cDNA libraries were screened with Aspergillus spp. and Saccharomyces cerevisiae aminopeptidase genes as the probes. Two leucine aminopeptidases, ruLap1 and ruLap2, and two dipeptidyl-peptidases, ruDppIV and ruDppV, were characterized and compared to orthologues secreted by Aspergillus fumigatus using a recombinant protein from Pichia pastoris. RuLap1 is a 33 kDa nonglycosylated protein, while ruLap2 is a 58-65 kDa glycoprotein. The hydrolytic activity of ruLap1, ruLap2 and A. fumigatus orthologues showed various preferences for different aminoacyl-7-amido-4-methylcoumarin substrates, and various sensitivities to inhibitors and cations. ruDppIV and ruDppV showed similar activities to A. fumigatus orthologues. In addition to endopeptidases, the four aminopeptidases ruLap1, ruLap2, ruDppIV and ruDppV were produced by T. rubrum in a medium containing keratin as the sole nitrogen source. Synergism between endo-and exopeptidases is likely to be essential for dermatophyte virulence, since these fungi grow only in keratinized tissues.

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“…High levels of these enzymes have been observed in plants grown under harsh conditions, especially under osmotic stress, which is due to water shortage or too high salinity (Bartling and Weiler 1992;Desimone et al 2000;Thayer et al 1988;Matsui et al 2006;Ogiwara et al 2005;Chien et al 2002). Therefore, they may be considered as an element of ''immunological response'' of plants (Fowler et al 2009;Bae et al 2013).…”

Section: Introductionmentioning

confidence: 99%

The influence of α-aminophosphonic acids on the activity of aminopeptidase from barley seeds—an approach to determine the enzyme specificity

2015

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Inhibitory potencies of 24 a-aminophosphonic acids against barley seeds (Hordeum vulgare L.) metalloaminopeptidase have been determined to evaluate structural requirements of this enzyme. The enzyme was sensitive mostly to the influence of phosphonic acid analogues of phenylalanine and its homologues, thus showing narrow specificity if compared with porcine aminopeptidases M1 and M17 and with Plasmodium aminopeptidase M17.

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Purification, characterization, and genetic analysis of a leucine aminopeptidase from Aspergillus sojae

Cited by 63 publications

References 41 publications

Extracellular phytase from Aspergillus niger CFR 335: purification and characterization

Extracellular phytase from Aspergillus niger CFR 335: purification and characterization

Aminopeptidases and dipeptidyl-peptidases secreted by the dermatophyte Trichophyton rubrum

The influence of α-aminophosphonic acids on the activity of aminopeptidase from barley seeds—an approach to determine the enzyme specificity

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