Compact archived record and sequence evidence for P15802
Gene Ontology annotation based on UniProtKB/Swiss-Prot keyword mapping
Gene Ontology annotation based on UniProtKB/Swiss-Prot Subcellular Location vocabulary mapping, accompanied by conservative changes to GO terms applied by UniProt.
Electronic Gene Ontology annotations created by ARBA machine learning models
Combined Automated Annotation using Multiple IEA Methods.
Cloning and heterologous expression of the penicillin biosynthetic gene cluster from penicillum chrysogenum.
-
Demonstrated heterologous expression of complete penicillin biosynthetic gene cluster from P. chrysogenum in N. crassa and A. niger
"Both of the transformed hosts contained intact P. chrysogenum DNA derived from the cosmid clone and produced authentic penicillin V"
-
Confirmed production of authentic penicillin V in transformed hosts
"Both of the transformed hosts contained intact P. chrysogenum DNA derived from the cosmid clone and produced authentic penicillin V"
-
Showed functional expression of acyl coenzyme A:6-aminopenicillanic acid acyltransferase (ACT) activity
"possessed delta-(L-alpha-amino-adipyl)-L-cysteinyl-D-valine synthetase (ACVS), isopenicillin N synthetase (IPNS) and acyl coenzyme A:6-aminopenicillanic acid acyltransferase (ACT) activity"
-
Established that all genes necessary for penicillin biosynthesis from amino acid precursors are closely linked in P. chrysogenum
"The data suggests that genes encoding all the enzymes necessary for the biosynthesis of penicillin from amino acid precursors are closely linked in P. chrysogenum and constitute a gene cluster"
The unprocessed preprotein form IATC103S of the isopenicillin N acyltransferase is transported inside peroxisomes and regulates its self-processing.
-
Demonstrated that unprocessed IAT(C103S) mutant is transported into peroxisomes, proving processing-independent transport
"Expression of a mutant penDE(C103S) gene in P. chrysogenum gave rise to an unprocessed inactive variant of IAT (IAT(C103S)) located inside peroxisomes"
-
Showed that IAT transport into peroxisomes does not require prior self-processing
"transport of the proIAT inside these organelles is not dependent on the processing state of the protein"
-
Discovered regulatory role of unprocessed IAT(C103S) in wild-type IAT processing
"Changes in the wild-type IAT processing profile (beta subunit formation) were observed in the Wis54-DE(C103S) strain, suggesting a regulatory role of the unprocessed IAT(C103S) in the processing of the wild-type IAT"
-
Confirmed post-translational regulation where preprotein interferes with self-processing
"Our results indicate that IAT is post-translationally regulated by its preprotein, which interferes with the self-processing"
Molecular characterization of the acyl-coenzyme A:isopenicillin N acyltransferase gene (penDE) from Penicillium chrysogenum and Aspergillus nidulans and activity of recombinant enzyme in Escherichia coli.
-
Cloned and characterized penDE gene encoding acyl-coenzyme A:isopenicillin N acyltransferase
"Based on NH2-terminal amino acid sequence information, the acyltransferase gene (penDE) from P. chrysogenum and A. nidulans were cloned"
-
Located penDE immediately downstream from isopenicillin N synthetase gene (pcbC)
"In both organisms, penDE was located immediately downstream from the isopenicillin N synthetase gene (pcbC)"
-
Identified gene structure with four exons encoding 357 amino acids (40 kDa protein)
"consisted of four exons encoding an enzyme of 357 amino acids (approximately 40 kilodaltons [kDa])"
-
Demonstrated functional expression in E. coli accepting both 6-aminopenicillanic acid and IPN as substrates
"Acyltransferase activity from Escherichia coli producing the 40-kDa protein accepted either 6-aminopenicillanic acid or IPN as the substrate and made a penicillinase-sensitive antibiotic in the presence of phenylacetyl coenzyme A"
-
Showed enzyme undergoes processing from 40 kDa precursor to active heterodimer with 11 and 29 kDa subunits
"The active form of the enzyme may result from processing of the 40-kDa monomeric precursor to a heterodimer containing subunits of 11 and 29 kDa"
Structures of an isopenicillin N converting Ntn-hydrolase reveal different catalytic roles for the active site residues of precursor and mature enzyme.
-
Solved crystal structure of Cys103Ala precursor mutant confirming autoproteolytic maturation
"The structure of a Cys103Ala precursor mutant shows that maturation is autoproteolytic"
-
Demonstrated that Cys103 initiates self-processing by cleaving its preceding peptide bond
"initiated by Cys103 cleaving its preceding peptide bond"
-
Revealed conformational change where residues 92-102 fold outward after autoproteolysis
"The crystal structure of the mature enzyme shows that after autoproteolysis residues 92-102 fold outwards, exposing a buried pocket"
-
Identified structurally flexible pocket that accommodates substrates of different size and polarity
"This pocket is structurally and chemically flexible and can accommodate substrates of different size and polarity"
-
Showed same catalytic residues perform different roles in autoproteolysis versus substrate hydrolysis
"Comparison of the proposed autoproteolytic and substrate hydrolysis mechanisms shows that in both events the same catalytic residues are used, but that they perform different roles in catalysis"
Deep research on aatA function
Acyl coenzyme A: 6-aminopenicillanic acid acyltransferase from Penicillium chrysogenum and Aspergillus nidulans.