X-Ray Crystallographic Study of a Non-Pepsin-Type Acid Proteinase, Aspergillus Niger Proteinase A

Sasaki, Hiroshi; Tanokura, Masaru; Muramatsu, Tsuyoshi; Nakagawa, Atsushi; Iwata, So; Hamaya, Toru; Takizawa, Toshio; Kono, Toshiaki; Takahashi, Kenji

doi:10.1007/978-1-4615-1871-6_81

Cited by 2 publications

(2 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In addition, we identified by MS an as yet uncharacterized putative glutamic endoprotease, AfuGprA (XP_748619, MER107323; encoding a gene at the locus AFUA_3G02970) (Monod et al, 2009;Sriranganadane et al, 2010), homologous to Aspergillus niger proteinase A or aspergillopepsin II (Takahashi, 2004) and to Scytalidium lignicolum scytalidopepsin B (Oda, 2004). A. niger aspergillopepsin II is not inhibited by pepstatin and was previously considered as a non-pepsin-type acid protease (Sasaki et al, 1995;Takahashi, 2004). The active site of scytalidopepsin B and aspergillopepsin II was identified with a catalytic dyad formed by residues Glu (E) and Gln (Q) (Fujinaga et al, 2004;Yabuki et al, 2004;Kataoka et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Secreted glutamic protease rescues aspartic protease Pep deficiency in Aspergillus fumigatus during growth in acidic protein medium

et al. 2011

View full text Add to dashboard Cite

In an acidic protein medium Aspergillus fumigatus secretes an aspartic endoprotease (Pep) as well as tripeptidyl-peptidases, a prolyl-peptidase and carboxypeptidases. In addition, LC-MS/MS revealed a novel glutamic protease, AfuGprA, homologous to Aspergillus niger aspergillopepsin II. The importance of AfuGprA in protein digestion was evaluated by deletion of its encoding gene in A. fumigatus wild-type D141 and in a pepΔ mutant. Either A. fumigatus Pep or AfuGprA was shown to be necessary for fungal growth in protein medium at low pH. Exoproteolytic activity is therefore not sufficient for complete protein hydrolysis and fungal growth in a medium containing proteins as the sole nitrogen source. Pep and AfuGprA constitute a pair of endoproteases active at low pH, in analogy to A. fumigatus alkaline protease (Alp) and metalloprotease I (Mep), where at least one of these enzymes is necessary for fungal growth in protein medium at neutral pH. Heterologous expression of AfuGprA in Pichia pastoris showed that the enzyme is synthesized as a preproprotein and that the propeptide is removed through an autoproteolytic reaction at low pH to generate the mature protease. In contrast to A. niger aspergillopepsin II, AfuGprA is a single-chain protein and is structurally more similar to G1 proteases characterized in other non-Aspergillus fungi.

show abstract

Section: Introductionmentioning

confidence: 99%

Secreted glutamic protease rescues aspartic protease Pep deficiency in Aspergillus fumigatus during growth in acidic protein medium

et al. 2011

View full text Add to dashboard Cite

show abstract

“…6) Our structural studies on microbial enzymes began with the X-ray crystallographic analysis of non-pepsintype acid proteinase from Aspergillus niger var. macrosporus 7,8) and ribonuclease T 1 (RNase T 1 ) from Aspergillus oryzae, 9) the latter of which specifically hydrolyzes a phosphate diester bond at the 3′-side of guanosine in single-stranded RNA. In subsequent reports, we have also revealed the structural basis of many enzymes catalyzing several reactions that have potentials for bioindustrial and/or bioremedial uses (Fig.…”

mentioning

confidence: 99%

Structural analysis of enzymes used for bioindustry and bioremediation

Tanokura

Miyakawa

Guan

et al. 2015

Bioscience, Biotechnology, and Biochemistry

Self Cite

View full text Add to dashboard Cite

Microbial enzymes have been widely applied in the large-scale, bioindustrial manufacture of food products and pharmaceuticals due to their high substrate specificity and stereoselectivity, and their effectiveness under mild conditions with low environmental burden. At the same time, bioremedial techniques using microbial enzymes have been developed to solve the problem of industrial waste, particularly with respect to persistent chemicals and toxic substances. And finally, structural studies of these enzymes have revealed the mechanistic basis of enzymatic reactions, including the stereoselectivity and binding specificity of substrates and cofactors. The obtained structural insights are useful not only to deepen our understanding of enzymes with potential bioindustrial and/or bioremedial application, but also for the functional improvement of enzymes through rational protein engineering. This review shows the structural bases for various types of enzymatic reactions, including the substrate specificity accompanying cofactor-controlled and kinetic mechanisms.

show abstract

X-Ray Crystallographic Study of a Non-Pepsin-Type Acid Proteinase, Aspergillus Niger Proteinase A

Cited by 2 publications

References 10 publications

Secreted glutamic protease rescues aspartic protease Pep deficiency in Aspergillus fumigatus during growth in acidic protein medium

Secreted glutamic protease rescues aspartic protease Pep deficiency in Aspergillus fumigatus during growth in acidic protein medium

Structural analysis of enzymes used for bioindustry and bioremediation

Contact Info

Product

Resources

About