| cloning and sequencing of a dextranase-encoding cdna from penicillium minioluteum. | a cdna from penicillium minioluteum hi-4 encoding a dextranase (1,6-alpha-glucan hydrolase, ec 3.2.1.11) was isolated and characterized. cdna clones corresponding to genes expressed in dextran-induced cultures were identified by differential hybridization. southern hybridization and restriction mapping analysis of selected clones revealed four different groups of cdnas. the dextranase cdna was identified after expressing a cdna fragment from each of the isolated groups of cdna clones in the esch ... | 1996 | 8837470 |
| cloning of the penicillium minioluteum gene encoding dextranase and its expression in pichia pastoris. | the dex gene encoding an extracellular dextranase was isolated from the genomic dna library of penicillium minioluteum by hybridization using the dextranase cdna as a probe. comparison of the gene and cdna sequences revealed that the dex gene does not contain introns. amino acid sequences comparison of p. minioluteum dextranase with other reported dextranases reveals a significant homology (29% identity) with a dextranase from arthrobacter sp. cb-8. the dex gene fragment encoding a mature protei ... | 1996 | 8905923 |
| molecular cloning and heterologous expression of the isopullulanase gene from aspergillus niger a.t.c.c. 9642. | isopullulanase (ipu) from aspergillus niger a.t.c.c. (american type culture collection) 9642 hydrolyses pullulan to isopanose. ipu is important for the production of isopanose and is used in the structural analysis of oligosaccharides with alpha-1,4 and alpha-1,6 glucosidic linkages. we have isolated the ipua gene encoding ipu from the filamentous fungi a. niger a.t.c.c. 9642. the ipua gene encodes an open reading frame of 1695 bp (564 amino acids). ipu contained a signal sequence of 19 amino ac ... | 1997 | 9169610 |
| structural model of dex protein from penicillium minioluteum and its implications in the mechanism of catalysis. | the dex gene encodes an extracellular dextranase (ec 3.2.1.11); this enzyme hydrolyzes the alpha(1,6) glucosidic bond contained in dextran to release small isomaltosaccharides. sequence analysis has revealed only one homologous sequence, cb-8 protein, from arthrobacter sp., with 30% sequence identity. the secondary structure prediction for dex was corroborated by circular dichroism measurements. to explore the possibility that dex protein might adopt a fold similar to any known structure, we con ... | 1998 | 9626695 |
| variation in n-linked oligosaccharide structures on heterologous proteins secreted by the methylotrophic yeast pichia pastoris. | we report the characterization of n-linked oligosaccharides on six foreign glycoproteins secreted from the methylotrophic yeast pichia pastoris. these proteins included: a bacterial enzyme, bacillus licheniformis alpha-amylase; three fungal enzymes, saccharomyces cerevisiae invertase, penicillium minioluteum dextranase, and mucor pusillus aspartic protease; and two higher eukaryotic proteins, boophilus microplus (tick) gut antigen and bovine enterokinase catalytic subunit. the carbohydrates on t ... | 1998 | 9790882 |
| be-31405, a new antifungal antibiotic produced by penicillium minioluteum. i. description of producing organism, fermentation, isolation, physico-chemical and biological properties. | a new antifungal antibiotic, be-31405, was isolated from the culture broth of a fungal strain, penicillium minioluteum f31405. be-31405 was isolated by adsorption on high porous polymer resin (diaion hp-20), followed by solvent extraction, precipitation and crystallization. be-31405 showed potent growth inhibitory activity against pathogenic fungal strains such as candida albicans, candida glabrata and cryptococcus neoformans, but did not show cytotoxic activity against mammalian cells such as p ... | 1998 | 10048566 |
| a novel fungal metabolite ng-061 enhances and mimics neurotrophic effect of nerve growth factor (ngf) on neurite outgrowth in pc12 cells. | during the course of our screening program for low molecular natural products with their ability to potentiate and/or mimic neurotrophic effect of ngf, a novel fungal metabolite, phenylacetic acid hydrazide derivative ng-061 was isolated from the fermentation broth of penicillium minioluteum f-4627. ng-061 enhanced and mimicked neurotrophic effect of ngf on neurite outgrowth in a rat pheochromocytoma cell line pc12. | 1999 | 10348036 |
| structure of ng-061, a novel potentiator of nerve growth factor (ngf) isolated from penicillium minioluteum f-4627. | the structure of ng-061, a new potentiator of nerve growth factor (ngf) isolated from penicillium minioluteum f-4627, was determined by spectroscopic analysis and x-ray diffraction method to be phenylacetic acid 2-(2-methoxy-4-oxocyclohexa-2,4-dienylidene)-hydrazide. | 1999 | 10348037 |
| carbon source regulation of a dextranase gene from the filamentous fungus penicillium minioluteum. | the regulation of dextranase (dexa) gene expression in the filamentous fungus penicillium minioluteum grown on different carbon sources was studied. growth in the presence of dextran leads to high expression of the dextranase enzyme, but growth in starch, glucose, glycerol, lactose and sorbitol did not. dextran induced dexa gene expression at the mrna level. however, in cultures containing dextran plus glucose or glycerol, the transcript was detected 24 h later than in the case where dextran was ... | 2000 | 10905430 |
| synthesis of ng-061 and its analogs, and their biological evaluation as an enhancer of nerve growth factor. | a novel potentiator of nerve growth factor (ngf), ng-061, which had been isolated from the fermentation broth of penicillium minioluteum f-4627, was synthesized from methoxybenzoquinone and phenylacetylhydrazine in a single step. a series of acyl hydrazone derivatives were also synthesized and their potentiator activity of neurotrophic effect of ngf on neurite outgrowth was evaluated by assay with a rat pheochromocytoma cell line pc12. | 2000 | 11045453 |
| new method for the selection of multicopy transformants of pichia pastoris, using 3-amino-1,2,4 triazol. | the methylotrophic yeast pichia pastoris has been successfully used for the expression of many heterologous proteins. the level of expression of some of these proteins depends on the copy number of the gene inserted into the yeast genome. several methods have been reported in the past few years for the isolation of multicopy transformants. one of these methods used an expression vector that contains the bacterial kanamycin-resistance gene tn903kanr, which confers resistance to g418. here, we rep ... | 2000 | 11084872 |
| molecular cloning of an alpha-glucosidase-like gene from penicillium minioluteum and structure prediction of its gene product. | the dexc cdna, which is expressed in dextran-containing medium by the filamentous fungus penicillium minioluteum, was cloned and sequence characterized. the cdna sequence comprises 1859 bp plus a poly (a) tail, coding for a predicted protein of 597 amino acids. the genomic counterpart was isolated by pcr, finding three introns in its sequence. the dexc gene was located by southern blot in the same 9-kb fragment that the previously isolated dextranase-encoding gene (dexa). sequence analysis revea ... | 2001 | 11178973 |
| preparation and crystallization of selenomethionyl dextranase from penicillium minioluteum expressed in pichia pastoris. | dextranase from the fungus penicillium minioluteum hydrolyses alpha-1,6-glycosidic bonds in dextran polymers. the enzyme has been expressed in pichia pastoris in the presence of selenomethionine (semet). the level of semet incorporation was estimated by amino-acid analysis to be 50%. the protein has been crystallized in space group p2(1)2(1)2, with unit-cell parameters a = 103.6, b = 115.3, c = 49.8 a and one molecule per asymmetric unit. the crystals diffract to 2.0 a and the presence of semet ... | 2002 | 11807273 |
| dextranase (alpha-1,6 glucan-6-glucanohydrolase) from penicillium minioluteum expressed in pichia pastoris: two host cells with minor differences in n-glycosylation. | differences in glycosylation between the natural alpha-1,6 glucan-6-glucanohydrolase from penicillium minioluteum and the heterologous protein expressed in the yeast pichia pastoris were analyzed. glycosylation profiling was carried out using fluorophore-assisted carbohydrate electrophoresis and amine absorption high-performance liquid chromatography (nh(2)-hplc) in combination with matrix-assisted laser desorption-time of flight-mass spectrometry. both microorganisms produce only oligomannosidi ... | 2001 | 12702360 |
| dextranase from penicillium minioluteum: reaction course, crystal structure, and product complex. | dextranase catalyzes the hydrolysis of the alpha-1,6-glycosidic linkage in dextran polymers. the structure of dextranase, dex49a, from penicillium minioluteum was solved in the apo-enzyme and product-bound forms. the main domain of the enzyme is a right-handed parallel beta helix, which is connected to a beta sandwich domain at the n terminus. in the structure of the product complex, isomaltose was found to bind in a crevice on the surface of the enzyme. the glycosidic oxygen of the glucose unit ... | 2003 | 12962629 |
| the icl1 gene of pichia pastoris, transcriptional regulation and use of its promoter. | we cloned and characterized a gene encoding isocitrate lyase from the methylotrophic yeast pichia pastoris. this gene was isolated from a p. pastoris genomic library using a homologous pcr hybridization probe, amplified with two sets of degenerate primers designed from conserved regions in yeast isocitrate lyases. the cloned gene was sequenced and consists of an open reading frame of 1563 bp encoding a protein of 551 amino acids. the molecular mass of the protein is calculated to be 60.6 kda wit ... | 2003 | 14558143 |
| insights into the reaction mechanism of glycosyl hydrolase family 49. site-directed mutagenesis and substrate preference of isopullulanase. | aspergillus niger isopullulanase (ipu) is the only pullulan-hydrolase in glycosyl hydrolase (gh) family 49 and does not hydrolyse dextran at all, while all other gh family 49 enzymes are dextran-hydrolysing enzymes. to investigate the common catalytic mechanism of gh family 49 enzymes, nine mutants were prepared to replace residues conserved among gh family 49 (four trp, three asp and two glu). homology modelling of ipu was also carried out based on the structure of penicillium minioluteum dextr ... | 2004 | 15560783 |
| cloning and characterization of a dextranase gene from lipomyces starkeyi and its expression in saccharomyces cerevisiae. | a dextranase-encoding cdna from l. starkeyi ksm22 was isolated and characterized. the 2052 bp cdna fragment (lsd1) harbouring the dextranase gene exhibited one open reading frame (orf) composed of 1824 bp flanked by a 41 bp 5'-utr and a 184 bp 3'-utr, including a 27 bp poly(a) tail. the lsd1 gene contains no introns. the open reading frame encodes a 608 amino acid polypeptide (lsd1) with a 67.6 kda predicted molecular mass. there was a 77% deduced amino acid sequence identity between the lsd1 de ... | 2005 | 16278932 |
| cloning and expression of penicillium minioluteum dextranase in saccharomyces cerevisiae and its exploitation as a reporter in the detection of mycotoxins. | a dextranase gene from penicillium minioluteum (strain imi068219) has been cloned, sequenced and expressed in saccharomyces cerevisiae via fusion of the dna segment encoding the mature dextranase protein with alpha-factor signal sequence, and insertion into the gal1-controlled expression vector pyes2/ct. galactose-induced expression yielded extracellular dextranase activity of 0.63 units/ml and cell-associated dextranase activity of 0.48 units/ml, after 24 h incubation. the dextranase construct ... | 2006 | 16988781 |
| pcr-rflp of its rdna for the rapid identification of penicillium subgenus biverticillium species. | rflp of its rdna is proposed as a useful tool for molecular identification of the most common species of biverticillate penicillia. 60 isolates were analysed representing 13 species and 21 unique sequences were produced. the combination of five restriction enzymes was successful in separating 12 species. however, the variety penicillium purpurogenum var. rubrisclerotium remained indistinguishable from penicillium funiculosum. p. funiculosum appeared as the most confused species, being mis-identi ... | 2006 | 17196020 |
| selenomethionine labeling of recombinant proteins. | selenomethionine incorporation is a standard method for determining the phases in protein crystallography by single- or multiwavelength anomalous dispersion. recombinant expression of selenomethionine-containing protein in non-auxotrophic pichia pastoris strains yield an incorporation of about 50%. the expression of a mutated variant of penicillium minioluteum dextranase in p. pastoris is used to illustrate the method utilized to obtain selenomethionyl-substituted protein and to show the phasing ... | 2007 | 17951642 |
| crystal structure of aspergillus niger isopullulanase, a member of glycoside hydrolase family 49. | an isopullulanase (ipu) from aspergillus niger atcc9642 hydrolyzes alpha-1,4-glucosidic linkages of pullulan to produce isopanose. although ipu does not hydrolyze dextran, it is classified into glycoside hydrolase family 49 (gh49), major members of which are dextran-hydrolyzing enzymes. ipu is highly glycosylated, making it difficult to obtain its crystal. we used endoglycosidase h(f) to cleave the n-linked oligosaccharides of ipu, and we here determined the unliganded and isopanose-complexed fo ... | 2008 | 18155243 |
| three new polyketide-terpenoid hybrids from penicillium sp. | three novel hybrid polyketide-terpenoid metabolites were isolated from a penicillium minioluteum strain. their structures were determined by nmr spectroscopic analyses and x-ray crystallography. the proposed biosynthetic pathway including a unique retro-claisen migration of methyl carbonate correlates the three compounds with berkeleydione and berkeleytrione. | 2008 | 18237180 |
| rational introduction of disulfide bond to enhance optimal temperature of lipomyces starkeyi alpha-dextranase expressed in pichia pastoris. | alpha-dextranase, which can hydrolyze dextran, is largely used in the sugar industry. however, a thermostable alpha- dextranase is needed to alleviate the viscosity of syrups and clean blocked machines. thus, to improve the optimal temperature of lipomyces starkeyi alpha-dextranase expressed by pichia pastoris, the rational introduction of a de novo designed disulfide bond was investigated. based on the known structure of penicillium minioluteum dextranase, l. starkeyi alpha-dextranase was const ... | 2009 | 20075611 |
| salinity stress resistance offered by endophytic fungal interaction between penicillium minioluteum lhl09 and glycine max. l. | endophytic fungi are little known for their role in gibberellins (gas) synthesis and abiotic stress resistance in crop plants. we isolated 10 endophytes from the roots of field-grown soybean and screened their culture filtrates (cf) on the gas biosynthesis mutant rice line - waito-c. cf bioassay showed that endophyte gmh-1b significantly promoted the growth of waito-c compared with controls. gmh-1b was identified as penicillium minioluteum lhl09 on the basis of its regions rdna sequence homology ... | 2011 | 21952365 |