Purification and Properties of Alkaline Proteinase from Aspergillus oryzae

Nakadai, Tadanobu; Nasuno, Seiichi; Iguchi, Nobuyoshi

doi:10.1271/bbb1961.37.2685

Cited by 19 publications

(12 citation statements)

References 1 publication

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“…A. jhmigatus secretes two other proteinases when the fungus is cultivated at neutral pH in the presence of collagen as the sole nitrogen and carbon source: a serine or alkaline proteinase (ALP) and a metalloproteinase (MEP) which are closely related to the alkaline proteinase and the neutral proteinase I secreted by A. olyzae, respectively [21,22]. However A. oryzae secretes a second metalloproteinase or neutral proteinase II [23], which appears to be absent in A. fumigatus.…”

Section: Resultsmentioning

confidence: 99%

Molecular cloning and sequencing of the gene encoding an extracellular aspartic proteinase fromAspergillus fumigatus

Reichard¹,

Monod

Rüchel³

1995

FEMS Microbiology Letters

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Oligonucleotide primers based on conserved regions of the aspergillopepsins (EC 3.4.23.18) were used to PCR amplify a 650 bp segment of the gene encoding the extracellular aspartic proteinase (PEP) from Aspergillus Fumigatus. The segment was used as a probe for isolating and sequencing the gene from a genomic library of the fungus. Likewise the cDNA was amplified by reverse PCR, cloned and sequenced. The pep gene was found to consist of four exons encoding for 395 aa. The pre-proenzyme deduced has an N-terminal leader sequence of 70 aa preceding the sequence of the mature enzyme consisting of 325 aa with a calculated molecular mass of 34.4 kDa and an isoelectric point of 3.95. The N-terminal sequence of the mature enzyme matched the N-terminal aa sequence of PEP exactly. The nucleotide and the aa sequences of the pre-proenzyme were 70% and 71% homologous to the corresponding sequences of the aspergillopepsin from A. niger var. awamori. Southern analysis of digested genomic A. fumigatus DNA with a specific PCR probe suggested the presence of a single copy of the pep gene.

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Section: Resultsmentioning

confidence: 99%

Molecular cloning and sequencing of the gene encoding an extracellular aspartic proteinase fromAspergillus fumigatus

Reichard¹,

Monod

Rüchel³

1995

FEMS Microbiology Letters

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“…Of these, alkaline, neutral and acidic proteases contribute significantly to the digestion of soybean protein during soy sauce brewing (Nakadai et al, 1973a, Nakadai et al, 1973b, Nakadai et al, 1973c, Nakadai and Nasuno, 1977). …”

Section: Construction Of An a Oryzae Quintuple Protease Gene Deletantmentioning

confidence: 99%

Construction of Quintuple Protease and Double Amylase Gene Deletant for Heterologous Protein Production in <i>Aspergillus oryzae</i> KBN616

Kitamoto¹,

Ono²,

Yoshino-Yasuda³

2015

FSTR

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We constructed a quintuple protease (alp, npII, pepE, npI, pepA) and double amylase (taaG3, taaG1) gene deletant, KO4, for the development of a heterologous gene expression system for an industrial shoyu koji mold, Aspergillus oryzae KBN616. Multiple gene deletion was performed using the Latour system, a simple and effective chromosome modification method developed in Schizosaccharomyces pombe. Gene deletion was confirmed by Southern blot analysis for protease genes and PCR amplification for amylase genes. Deletion of the five protease genes reduced the extracellular protease activity to approximately 1.0% of the level of the parent strain. Double deletion of the amylase genes completely eliminated all detectable α-amylase activity. These results indicate the construction of a host strain applicable to the efficient production and purification of heterologous proteins.

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“…Aspergillus candidus [13] A. flavus [14] A. fumigatus [15,16] A. melleus [17] A. niger [18,19] A. oryzae [20][21][22] A. sojae [23] A. sydowi [24] Cephalosporium sp. KSM 388 [25] Chrysosporiumkeratinophilum [26] Conidioboluscoronatus [27] Entomophthoracoronata [28] Fusariumeumartii [29] Paecilomyces lilacinus [30] Scedosporium apiospermum [31] Tritirachium album Limber [32][33][34] Rhizopusoligosporus [35] Table 3.…”

Section: Fungus Referencesmentioning

confidence: 99%

“…Recent years have witnessed a phenomenal increase in the use of enzymes as industrial catalysts. Proteases (EC 3:4,(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)99) (synonymous as peptidase or proteinase) constitute a very large and complex group of enzymes, widely utilized in a host of industries. They differ in properties such as substrate specificity, active site and catalytic mechanism, pH and temperature optima, and stability profiles.…”

Section: Introductionmentioning

confidence: 99%

Versatility of microbial proteases

Veloorvalappil¹,

Robinson²,

Pradeep³

et al. 2013

AER

110

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Proteases or peptidases constitute the largest group of enzymes in bio-industry with a long array of uses. They play an invincible role in industrial biotechnology, especially in detergent, food and pharmaceutical arena. This focused review encompasses an overview on alkaline proteases, mainly of microbial sources in a handy module. Following an introduction and general classification with evolutionary insight, major sources of proteases (animal, plant and microbial including fungal, bacterial), their general properties with mechanism of action and molecular masses are discussed. Proteases from Bacillus spp. have been given special attention. In addition to this, an overview on the applications of proteases in detergent, tannery, food, metal recovery and waste treatment industries is also addressed briefly.

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Purification and Properties of Alkaline Proteinase from Aspergillus oryzae

Cited by 19 publications

References 1 publication

Molecular cloning and sequencing of the gene encoding an extracellular aspartic proteinase fromAspergillus fumigatus

Molecular cloning and sequencing of the gene encoding an extracellular aspartic proteinase fromAspergillus fumigatus

Construction of Quintuple Protease and Double Amylase Gene Deletant for Heterologous Protein Production in <i>Aspergillus oryzae</i> KBN616

Versatility of microbial proteases

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