Purification and Characterization of a Novel Prolyl Aminopeptidase from<i>Maitake</i>(<i>Grifola frondosa</i>)

Hiwatashi, Koichi; Hori, Kazuyuki; Takahashi, Keitaro; Kagaya, Akira; Inoue, Shunzo; Sugiyama, Toshihiro; Takahashi, Saori

doi:10.1271/bbb.68.1395

Cited by 14 publications

(4 citation statements)

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“…In addition, products of pepN , pepS and pepX genes are aminopeptidases that need a free N‐terminus and cleave off either one or two amino acids from the end of a peptide . Up‐regulation of L. sakei pepX in meat extract would reduce bitter tastes by degrading proline‐containing peptides once they have been transported inside the cell or after the release of the intracellular enzymes to the meat matrix by cell lysis . Genes pepD1 and pepD5 (encoding U34 family dipeptidases pepD1 and pepD5) are cysteine‐type, N‐terminal nucleophile (Ntn)‐hydrolases, which are remote homologs of penicillin V acylases, sharing similar catalytic machinery but with a sequence diversity enabling the catalysis of reactions involving hydrolysis of amid bonds .…”

Section: Discussionmentioning

confidence: 99%

Transcriptome response ofLactobacillus sakeito meat protein environment

Yun-shen

Tian

et al. 2014

J. Basic Microbiol.

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Lactobacillus sakei is a heterofermentative species of lactic acid bacteria that is used in industrial meat fermentation. To investigate adaptation in a meat environment, whole-genome DNA microarrays were used to analyze the gene expression related to growth and survival of L. sakei strain La22 when grown in sarcoplasmic (S-) or myofibrillar (M-) protein-supplemented chemically defined medium (CDM). Differential expression was detected in 551 genes. Genes encoding enzymes involved in peptide hydrolysis were differentially upregulated in M-CDM or/and S-CDM, and only oppB and oppC, involved in the amino acid and peptide transport system, were upregulated. Most genes related to metabolism of peptides, amino acids and related molecules were over-expressed in M-CDM and S-CDM, except for glnA and metK. Expression of certain genes was according to the differential substrate environment. The expression of genes involved in the stress response was not induced by growth in M-CDM.

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Section: Discussionmentioning

confidence: 99%

Transcriptome response ofLactobacillus sakeito meat protein environment

Yun-shen

Tian

et al. 2014

J. Basic Microbiol.

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“…Controlled enzymatic debittering using aminopeptidases can be both economic and effective (Saha and Hayashi, 2001). To date, bacterial and fungal aminopeptidases such as leucine aminopeptidases, prolyl aminopeptidase and alanyl aminopeptidase, have been used in protein hydrolysate debittering (FitzGerald and O'cuinn, 2006;Hiwatashi et al, 2004;Huang et al, 2015;Mane et al, 2010;Lin et al, 2004;Nampoothiri et al, 2005;Stressler et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

A novel aminopeptidase with potential debittering properties in casein and soybean protein hydrolysates

Song

Cheng

Tian

et al. 2020

Food Sci Biotechnol

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A new aminopeptidase (An-APa) was identified and biochemically characterized from Aspergillus niger CICIM F0215. It had maximal activity at 40 °C and pH 7.0 and exhibited a broad substrate specificity both on hydrophilic and hydrophobic amino acid residues at N-terminals. With An-APa hydrolysis for 1 h, the casein-pepsin and soybean protein isolates (SPI)-pepsin hydrolysates released both hydrophilic and hydrophobic amino acids and the hydrophobic amino acids having Q values (degree of hydrophobicity) greater than 1500 cal/mol were remarkably released.

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“…A number of PAP-type activities in fungi have been reported (Bolumar et al, 2003;Fuke & Matsuoka, 1993;Hiwatashi et al, 2004), but no corresponding protein sequences have been assigned for these enzymes. However, more recently, one study by Basten et al (2005) identified a gene encoding PapA from Aspergillus niger and characterized the recombinant protein.…”

Section: Introductionmentioning

confidence: 99%

Characterization of a multimeric, eukaryotic prolyl aminopeptidase: an inducible and highly specific intracellular peptidase from the non-pathogenic fungus Talaromyces emersonii

et al. 2009

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Fungi are capable of degrading proteins in their environment by secreting peptidases. However, the link between extracellular digestion and intracellular proteolysis has scarcely been investigated. Mycelial lysates of the filamentous fungus Talaromyces emersonii were screened for intracellular peptidase production. Five distinct proteolytic activities with specificity for the p-nitroanilide (pNA) peptides Suc-AAPF-pNA, Suc-AAA-pNA, K-pNA, F-pNA and P-pNA were identified. The native enzyme responsible for the removal of N-terminal proline residues was purified to homogeneity by ammonium sulfate fractionation followed by five successive chromatographic steps. The enzyme, termed Talaromyces emersonii prolyl aminopeptidase (TePAP), displayed a 50-fold specificity for cleaving N-terminal Pro-X (k cat /K m 52.1¾10 6 M "1 s "1 ) compared with Ala-X or Val-X bonds. This intracellular aminopeptidase was optimally active at pH 7.4 and 50 6C. Peptide sequencing facilitated the design of degenerate oligonucleotides from homologous sequences encoding putative fungal proline aminopeptidases, enabling subsequent cloning of the gene. TePAP was shown to be relatively uninhibited by classical serine peptidase inhibitors and to be sensitive to selected cysteine-and histidine-modifying reagents, yet gene sequence analysis identified the protein as a serine peptidase with an a/b hydrolase fold. Northern analysis indicated that Tepap mRNA levels were regulated by the composition of the growth medium. Highest Tepap transcript levels were observed when the fungus was grown in medium containing glucose and the protein hydrolysate casitone. Interestingly, both the induction profile and substrate preference of this enzyme suggest potential co-operativity between extracellular and intracellular proteolysis in this organism. Gel filtration chromatography suggested that the enzyme exists as a 270 kDa homo-hexamer, whereas most bacterial prolyl aminopeptidases (PAPs) are monomers. Phylogenetic analysis of known PAPs revealed two diverse subfamilies that are distinguishable on the basis of primary and secondary structure and appear to correlate with the subunit composition of the native enzymes. Sequence comparisons revealed that PAPs with key conserved topological features are widespread in bacterial and fungal kingdoms, and this study identified many putative PAP candidates within sequenced genomes. This work represents, to our knowledge, the first detailed biochemical and molecular analysis of an inducible PAP from a eukaryote and the first intracellular peptidase isolated from the thermophilic fungus T. emersonii.Abbreviations: AEBSF, 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride; DEPC, diethylpyrocarbonate; E-64, trans-epoxy-succinyl-L-leucylamido-(4-guanidino)-butane; NEM, N-ethylmaleimide; PAP, prolyl aminopeptidase; PCMB, p-chloromercuribenzoate; pNA, p-nitroanilide; Pro-AMC, L-proline-7-amino-4-methylcoumarin; TLCK, N-tosyl-L-lysine chloromethyl ketone; TPCK, N-tosyl-L-phenylalanine chloromethyl ketone.3These authors contrib...

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Purification and Characterization of a Novel Prolyl Aminopeptidase fromMaitake(Grifola frondosa)

Cited by 14 publications

References 10 publications

Transcriptome response ofLactobacillus sakeito meat protein environment

Transcriptome response ofLactobacillus sakeito meat protein environment

A novel aminopeptidase with potential debittering properties in casein and soybean protein hydrolysates

Characterization of a multimeric, eukaryotic prolyl aminopeptidase: an inducible and highly specific intracellular peptidase from the non-pathogenic fungus Talaromyces emersonii

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