• Journal of Microbiology and Biotechnology
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• v.20 no.5
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• pp.893-903
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• 2010

A cellulolytic and xylanolytic enzyme complex-producing alkalothermoanaerobacterium strain, Tepidimicrobium xylanilyticum BT14, is described. The cell was Grampositive, rod-shaped, and endospore-forming. Based on 16S rRNA gene analysis and various lines of biochemical and physiological properties, the strain BT14 is a new member of the genus Tepidimicrobium. The strain BT14 cells had the ability to bind to Avicel, xylan, and corn hull. The pH and temperature optima for growth were 9.0 and $60^{\circ}C$, respectively. The strain BT14 was able to use a variety of carbon sources. When the bacterium was grown on corn hulls under an anaerobic condition, a cellulolytic and xylanolytic enzyme complex was produced. Crude enzyme containing cellulase and xylanase of the strain BT14 was active in broad ranges of pH and temperature. The optimum conditions for cellulase and xylanase activities were pH 8.0 and 9.0 at $60^{\circ}C$, respectively. The crude enzyme had the ability to bind to Avicel and xylan. The analysis of native-PAGE and native-zymograms indicated the cellulosebinding protein showing both cellulase and xylanase activities, whereas SDS-PAGE zymograms showed 4 bands of cellulases and 5 bands of xylanases. Evidence of a cohesinlike amino acid sequence seemed to indicate that the protein complex shared a direct relationship with the cellulosome of Clostridium thermocellum. The crude enzyme from the strain BT14 showed effective degradation of plant biomass. When grown on corn hulls at pH 9.0 and $60^{\circ}C$ under anaerobic conditions, the strain BT14 produced ethanol and acetate as the main fermentation products.

• The Korean Journal of Mycology
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• v.27 no.5 s.92
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• pp.328-336
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• 1999

Isozyme comparisons of mycelial extracts from Pleurotus ostreatus were undertaken using isoelectric focusing. Enzyme isozyme patterns were Used to describe the extent of geographical diversity and degree of intraspecific variation in these extracts. A total of 77 bands were resolved from six different enzymes. Cluster analyses were performed using the zymograms for esterase (EST), glucose phosphate isomerase (GPI), leucine aminopeptidase (LAP), malate dehydrogenase(MDH), peroxidase (POX), and phosphoglucomutase (pGM). EST gave multiple banding patterns, while less variability was observed for GPI, MDH, and PGM. Cluster analyses demonstrated that strains of P. ostreatus from geographically different origins are genetically divergent, supporting the idea that there is little or no gene flow between these geographically distant population groups.

• Journal of Veterinary Clinics
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• v.21 no.3
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• pp.248-252
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• 2004

DNase activity from infective larvae of the parasitic nematode Haemonchus contortus was characterized and compared to that from whole worm. DNase activity from infective larvae was detected throughout pHs 4-10, but high activity was detected under acidic conditions. The activity was not inhibited by 10 mM EDTA at pH 5.0, but was significantly inhibited at pH 7.0. The activity produced DNA, fragments with mixtures of 3'-hydroxyls (OH) and 3'-phosphates (P) at each pH with predominance of 3'-P. A unique DNase activity at 37 kDa was identified from infective larvae on zymograms. The 37 kDa DNase was detected only at pH 5.0, but not at pH 7.0, and this activity was not inhibited by EDTA at pH 5.0. These characteristics of the 37 kDa infective larval DNase resemble those of classic acidic DNases (e.g., DNase II). In contrast, 34, 36 and 38.5 kDa DNase activities were shown to be specific for whole worm. This result demonstrated that DNases in H contortus are regulated during development.

• Journal of The Korean Society of Grassland and Forage Science
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• v.15 no.2
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• pp.106-111
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• 1995

This study was planned to identify the effect of low-temperature stress on Alcohol dehydrogenase(ADH) isozyme in sixteen varieties of Italian ryegrass using starch gel electrophoresis. The specific electrophoretic zymograms of each variety were observed by ADH isozyme. The results were summarized as follows: 1. All tested varieties displayed two band zone by ADH and R.f values were 0.63 and 0.60, respectively. 2. There were four band type for ADH isozyme of 16 varieties classified with ADH isozyme dyeing intensity. According to dyeing intensity 7, 2, 1 and 6 varieties belong to banding type I,II,III and IV, respectively(Fig.2-A, B). 3. The effect of short tern low-temperature stress induces ADH gene expresson in Italian ryegrass, which may reflect a fundmental shift in energy metabolism to ensure plant tissue survival during the low-temperature stress period.

• The Korean Journal of Zoology
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• v.19 no.1
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• pp.33-42
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• 1976

The blood proteins of ten reptilian species were studied by cellulose acetate elctrophoresis. Three members examined of the genus Agkistrodon have unusually similar patterns in plasma protein, hemoglobin, lactate dehydrogenase and malate dehydrogenase. On the basis of their electrophoretic patterns, it was concluded that Agkistrodon blomhoffii brevicaudus was closely related to the others. In general the plasma protein patterns reflect species specificity. Under the conditions employed, all snakes had a single hemoglobin band except Dinodon rufozonatum rufozonatum which showing two component patterns. Two members of the Chelonia showed four bands of hemoglobin. The zymograms indicated a distinct divergence in blood proteins of the Squamata from those of the Chelonia. The results reflected superficially the established phylogenies of these groups.

• The Korean Journal of Mycology
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• v.26 no.2 s.85
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• pp.163-172
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• 1998

Thirty six strains of Pleurotus spp., from world-wide nations, were examined for interspecific isozyme variation. A comparison of isozymes in mycelial extracts of the fungal genus Pleurotus was made by polyacrylamide gel isoelectric focusing. A total of one hundred and sixty six bands was resolved from six isozymes. A cluster analysis was done based on the zymograms for esterase, glucosephosphate isomerase, leucine aminopeptidase, malate dehydrogenase, peroxidase, and phosphoglucomutase. From the isozyme analysis, esterase showed higher degree of variability, while it was observed less variability for the enzymes such as glucosephosphate isomerase, malate dehydrogenase, and phosphoglucomutase. The species P. ostreatus, whose taxon is controversial, was discriminated from P. pulmonarius, while P. florida was classified as a distinct taxon. The clustering of P. sapidus and P. spodoleucus strains appeared to be more difficult. It was found that some strains were included to another cluster based on electrophoretic banding patterns. These results show that this lack of congruence among data sets may help explain the taxonomic difficulty within the genus Pleurotus. A dendrogram of genetic similarities was presented, and applications of isozyme data to the systematics of these commercially important fungi was discussed.

• Applied Biological Chemistry
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• v.14 no.3
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• pp.229-235
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• 1971

Kwanok, Fujisaka #5, Paldal, and Suwon #82 as japonica type and IR-262 and CP-slo as indica type of rice seeds were selected for this experiment among varieties grown in Korea. Activities of crude enzymes extracted from germinating seeds of these varieties on malathion and p-nitrophenyl acetate as substrates, esterase zymograms with 1-naphthyl acetate as substrate, and some observations are summarized as follows: 1. Activities per unit volume of crude enzyme preparations on malathion were in the order of Kwanok>IR-262>Fujisaka #5>CP-slo>Paldal>Suwon #82. 2. Esterase zymograms on agar-gel electrophoretograms exhibited three to four bands two electrodes with little difference among varieties, nevertheless showing a wide and strongly-colored band toward cathode. Suwon #82 has a somewhat different pattern from others. 3. Enzyme activities per milligram protein with p-nitrophenyl acetate as substrate were in the order of CP-slo>IR-262>Paldal>Kwanok>Suwon #82>Fujisaka #5, indicating that activities of indica type are much stronger than those of japonica type, but not in agreement with results with malathion. 4. Malathion did not much inhibit the esterase activity at the concentration of 0.2PPM on electrophoretograms. 5. It is supposed that there is a complex esterase system hydrolyzing malathion and p-nitrophenyl acetate in germinating rice seeds.

• BMB Reports
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• v.44 no.3
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• pp.193-198
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• 2011

The chitinase-producing strain SC081 was isolated from Korean traditional soy sauce and identified as Bacillus atrophaeus based on a phylogenetic analysis of the 16S rDNA sequence and a phenotypic analysis. A gene encoding chitinase from B. atrophaeus SC081 was cloned in Escherichia coli and was named SCChi-1 (GQ360078). The SCChi-1 nucleotide sequences were composed of 1788 base pairs and 596 amino acids, which were 92.6, 89.6, 89.3, and 78.9% identical to those of Bacillus subtilis (ABG57262), Bacillus pumilus (ABI15082), Bacillus amyloliquefaciens (ABO15008), and Bacillus licheniformis (ACF40833), respectively. A recombinant SCChi-1 containing a hexahistidine tag at the amino-terminus was constructed, overexpressed, and purified in E. coli to characterize SCChi-1. $H_6SCChi$-1 revealed a hydrolytic band on zymograms containing 0.1% glycol chitin and showed the highest lytic activity on colloidal chitin and acidic chitosan. The optimal temperature and pH for chitinolytic activity were $50^{\circ}C$ and pH 8.0, respectively.

• Mycobiology
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• v.35 no.4
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• pp.219-225
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• 2007

A keratinolytic enzyme secreted by Aspergillus flavus K-03 cultured in feather meal basal medium (FMBM) containing 2% (w/v) chicken feather was purified and characterized. Keratinolytic enzyme secretion was the maximal at day 16 of the incubation period at pH 8 and $28^{\circ}C$. No relationship was detected between enzyme yield and increase of fungal biomass. The fraction obtained at 80% ammonium sulfate saturation showed 2.39-fold purification and was further purified by gel filtration in Sephadex G-100 followed by ion exchange chromatography on DEAE-Sephadex A-50, yielding an active protein peak showing 11.53-fold purification. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and zymograms indicated that the purified keratinase is a monomeric enzyme with 31 kDa molecular weight. The extracellular keratinase of A. flavus was active in a board range of pH ($7{\sim}10$) and temperature ($30^{\circ}C{\sim}70^{\circ}C$) profiles with the optimal for keratinase activity at pH 8 and $45^{\circ}C$. The keratinase activity was totally inhibited by protease inhibitors such as phenylmethylsulfonyl fluoride (PMSF), iodoacetic acid, and ethylenediaminetetraacetate (EDTA) while no reduction of activity by the addition of dithiothreitol (DTT) was observed. N-terminal amino acid sequences were up to 80% homologous with the fungal subtilisins produced by Fusarium culmorum. Therefore, on the basis of these characteristics, the keratinase of A. flavus K-03 is determined to be subtilisins-like.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2001.02a
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• pp.55-56
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• 2001

Gelatin zymograms of bFF and bS showed GA110 and 62 kDa gelatinses in adsition to several minor ones. Of these, GA110 gelatinase was abolished by treating bFF or bS with bOF and interestingly, its enzymatic activity was enhanced by adding EDTA to bFF or bS before zymographic analyses. Experiments using specific inhibitors of MMPs indicated that GA110 and 62 kDa proteins were indeed gelatinases. Immunoblotting experiments using an antibody against human MMP-2 showed that both GA110 and 62 kDa were an MMP-2 isoform and active MMP-2, respectively. The results suggest that the interaction between bFF and bOF can occur at the time of fertilization.