• KSBB Journal
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• 제10권3호
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• pp.335-342
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• 1995

농산 폐기물인 배추 뿌리에 다량으로 존재하는 peroxidase를 산업적으로 이용하기 위하여 배추 뿌 라를 JUicer로 대량 추출하여 객상효소 부분과 고형 물 부분을 얻었다. Peroxidase는 액상 부분에 약 6 66%, 고형물 부분에 34%가 분포되어 있으므로 두 부분을 모두 이용하여 phenol성 폐수의 효소적 처리를 검토하였다. Batch stirred reactor에서 액상효소(1,800 unit/$\ell$) 를 이용하여 150ppm의 phenol 용액을 처리한 결과 3시간 후에 96% 의 phenol을 중 합시켜 침전으로 제거할 수 있었다. 한편 pulp를 이용한 air lift reactor(600 unit/$\ell$) 에서는 120ppm의 초기 phenol 농도로부터 5ppm까지 제거할 수 있 다. Batch stirred reactor에 비하여 air lift reactor에 첨가된 효소의 양이 1/3임에도 불구하고 거의 비숫한 phenol 제거 효율을 냐타내었다.

• Bulletin of the Korean Chemical Society
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• 제15권9호
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• pp.719-724
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• 1994

Major cellulase components, such as three endoglucanases (endoglucanases I, II, and III) and one exoglucanase (exoglucanase II), were isolated from a commercial cellulase (Meicelase TP 60) derived from the fungus Trichoderma viride by a series of chromatography procedures. These procedures were the gel filtration on Bio-Gel, the anion exchange on DEAE-Bio-Gel A, the cation exchange on SP-Sephadex C50, and the affinity chromatography on Avicel cellulose. The average molecular weights determined by SDS-polyacrylamide gel electrophoretic analysis were 51,000, 59,000, 41,000 and 62,000 Da for endoglucanases I, II and III and exoglucanase II, respectively. The extinction coefficients, ${\varepsilon}^{1%}$ 280 nm, of these enzymes were 11.7, 3.3, 7.2 and 11.3, respectively. Among them, the endoglucanase II showed the very low value of the coefficient compared with the others. On the other hand, it was found that endoglucanase II and III were of more random hydrolytic mode on carboxymethylcellulose as compared with those of endoglucanase I and exoglucanase II. Especially, endoglucanase I showed less random action than that of exoglucanase II. In the hydrolysis of insoluble cellulose by the enzyme components, cellobiose was the major product, but glucose was the major product by endoglucanase III.

• 한국해양생명과학회지
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• 제7권2호
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• pp.121-128
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• 2022

L-asparaginase (ASNase) is a therapeutic enzyme used to treat acute lymphoblastic leukemia. Currently, the most widely used ASNases are originated from bacteria. However, owing to the adverse effects of bacterial ASNases, new resources for ASNase production should be explored. Fungal enzymes are considered efficient and compatible resources of natural products for diverse applications. In particular, fungal species belonging to the genus Trichoderma are well-known producers of several commercial enzymes including cellulase, chitinase, and xylanase. However, enzyme production by marine-derived Trichoderma spp. remains to be elucidated. While screening for extracellular ASNase-producing fungi from marine environments, we found four strains showing extracellular ASNase activity. Based on the morphological and phylogenetic analyses using sequences of translation elongation factor 1-alpha (tef1α), the Trichoderma isolates were identified as T. afroharzianum, T. asperellem, T. citrinoviride, and Trichoderma sp. 1. All four strains showed different ASNase activities depending on the carbon sources. T. asperellem MABIK FU00000795 showed the highest ASNase value with lactose as a carbon source. Based on our findings, we propose that marine-derived Trichoderma spp. are potential candidates for novel ASNase production.

• Journal of Microbiology
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• 제45권6호
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• pp.485-491
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• 2007

The effects of biological pretreatment on the Japanese red pine Pinus densiflora, was evaluated after exposure to three white rot fungi Ceriporia lacerata, Stereum hirsutum, and Polyporus brumalis. Change in chemical composition, structural modification, and their susceptibility to enzymatic saccharification in the degraded wood were analyzed. Of the three white rot fungi tested, S. hirsutum selectively degraded the lignin of this sortwood rather than the holocellulose component. After eight weeks of pretreatment with S. hirsutum, total weight loss was 10.7%, while lignin loss was the highest at 14.52% among the tested samples. However, holocellulose loss was lower at 7.81 % compared to those of C. lacerata and P. brumalis. Extracelluar enzymes from S. hirsutum showed higher activity of ligninase and lower activity of cellulase than those from other white rot fungi. Thus, total weight loss and changes in chemical composition of the Japanese red pine was well correlated with the enzyme activities related with lignin- and cellulose degradation in these fungi. Based on the data obtained from analysis of physical characterization of degraded wood by X-ray Diffractometry (XRD) and pore size distribution, S. hirsutum was considered as an effective potential fungus for biological pretreatment. In particular, the increase of available pore size of over 120 nm in pretreated wood powder with S. hirsutum made enzymes accessible for further enzymatic saccharification. When Japanese red pine chips treated with S. hirsutum were enzymatically saccharified using commercial enzymes (Cellulclast 1.5 L and Novozyme 188), sugar yield was greatly increased (21.01 %) compared to non-pre treated control samples, indicating that white rot fungus S. hirsutum provides an effective process in increasing sugar yield from woody biomass.

• 생명과학회지
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• 제16권2호
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• pp.360-364
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• 2006

Fibrin clots of blood vessels are one of the serious factor caused cardiovascular disease. The development of a antithrombotic and thrombolysis solvent is necessary to prevent and treat these diseases. It has been reported that a strong fibrin-specific fibrinolytic enzyme was produced from a Korean fermented soybean paste similar to Japanese miso. We have been screened the known or novel fibrinolytic enzymes by activity-based and sequence-based screening from soil DNA metagenome library containing all kinds of environmental genomic DNA. The activity-based screening was determined the protease activity on 0.5% skim milk. For sequence-based screening, we designed a set of primer expanding gene sequence of fibrinolytic enzyme, performed PCR and selected clones showing the expected size of amplicons from metagenome library. Transformation of the gene encoding fibrinolytic enzyme was carried out with commercial vectors and their transformants were selected. Finally, we found 15 positive clones from metagenome library. Then each of sequences were analyzed and identified as similar or known the clones of nattokinase. We are going to perform full sequence of each clones, ligate with expression vector, transform into competent cells and then determine activity of expressed enzymes.

• Asian-Australasian Journal of Animal Sciences
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• 제21권1호
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• pp.97-102
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• 2008

Four hundred and forty 1-day-old Arbor Acre broilers were fed commercial starter diets with 0, 250, 750 and 2,250 mg/kg of an alpha-amylase preparation from 1 to 21 days of age to investigate the effects of an exogenous enzyme on growth performance, activities of digestive enzymes in the pancreas and anterior intestinal contents and pancreatic amylase mRNA expression. Body weight gain (BWG) and average daily gain (ADG) increased linearly (p<0.01) with increasing levels of supplementary amylase but feed conversion ratio (FCR) was not affected. There was a negative quadratic change of protease and amylase in the small intestinal contents with the increase of supplementary amylase level. The activity of intestinal trypsin was also increased (p<0.05). Lipase was unaffected by amylase supplementation (p>0.05). The pancreatic protease, trypsin, and lipase were not affected by exogenous amylase levels. Consistent with the tendency for a linear depression of amylase activity, pancreatic ${\alpha}$-amylase mRNA was down-regulated by dietary amylase supplementation. The present study suggested that oral administration of exogenous amylase affected activities of intestinal enzymes and the production of pancreatic digestive enzymes in a dose-dependent manner.

• Journal of Microbiology and Biotechnology
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• 제19권11호
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• pp.1306-1318
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• 2009

The filamentous ascomycete Sclerotinia sclerotiorum is well known for its ability to produce a large variety of hydrolytic enzymes. Two $\alpha$-amylases ScAmy54 and ScAmy43 predicted to play an important role in starch degradation were showed to produce specific oligosaccharides essentially maltotriose that have a considerable commercial interest. Primary structure of the two enzymes was established by N-terminal sequencing, MALDI-TOF masse spectrometry and cDNA cloning. The two proteins have the same N-terminal catalytic domain and ScAmy43 derived from ScAmy54 by truncation of 96 amino acids at the carboxyl-terminal region. Data of genomic analysis suggested that the two enzymes originated from the same $\alpha$-amylase gene and that truncation of ScAmy54 to ScAmy43 occurred probably during S. sclerotiorum cultivation. The structural gene of Scamy54 consisted of 9 exons and 8 introns, containing a single 1,500-bp open reading frame encoding 499 amino acids including a signal peptide of 21 residues. ScAmy54 exhibited high amino acid homology with other liquefying fungal $\alpha$-amylases essentially in the four conserved regions and in the putative catalytic triad. A 3D structure model of ScAmy54 and ScAmy43 was built using the 3-D structure of 2guy from A. niger as template. ScAmy54 is composed by three domains A, B, and C, including the well-known $(\beta/\alpha)_8$ barrel motif in domain A, have a typical structure of $\alpha$-amylase family, whereas ScAmy43 contained only tow domains A and B is the first fungal $\alpha$-amylase described until now with the smallest catalytic domain.

• The Plant Pathology Journal
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• 제31권1호
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• pp.50-60
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• 2015

The use of novel isolates of Trichoderma with efficient antagonistic capacity against Fusarium oxysporum f. sp. lycopersici (FOL) is a promising alternative strategy to pesticides for tomato wilt management. We evaluated the antagonistic activity of 30 isolates of T. asperellum against 4 different isolates of FOL. The production of extracellular cell wall degrading enzymes of the antagonistic isolates was also measured. The random amplified polymorphic DNA (RAPD) method was applied to assess the genetic variability among the T. asperellum isolates. All of the T. asperellum isolates significantly reduced the mycelial growth of FOL isolates but the amount of growth reduction varied significantly as well. There was a correlation between the antagonistic capacity of T. asperellum isolates towards FOL and their lytic enzyme production. Isolates showing high levels of chitinase and ${\beta}$-1,3-glucanase activities strongly inhibited the growth of FOL isolates. RAPD analysis showed a high level of genetic variation among T. asperellum isolates. The UPGMA dendrogram revealed that T. asperellum isolates could not be grouped by their antagonistic behavior or lytic enzymes production. Six isolates of T. asperellum were highly antagonistic towards FOL and potentially could be used in commercial agriculture to control tomato wilt. Our results are consistent with the conclusion that understanding the genetic variation within Trichoderma isolates and their biochemical capabilities are required for the selection of effective indigenous fungal strains for the use as biocontrol agents.

• Journal of Microbiology
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• 제43권6호
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• pp.493-498
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• 2005

The safety assessment of Bifidobacterium longum SPM1205 isolated from healthy Koreans and this strain's inhibitory effects on fecal harmful enzymes of intestinal microflora were investigated. The overall safety of this strain was investigated during a feeding trial. Groups of SD rats were orally administered a test strain or commercial reference strain B. longum $1\times10^9\;CFU/kg$ body weight/day for four weeks. Throughout this time, their feed intake, water intake and live body weight were monitored. Fecal samples were periodically collected to test harmful enzyme activities of intestinal microflora. At the end of the four-week observation period, samples of blood, liver, spleen, kidney, and gut tissues were collected to determine for hematological parameters and histological differences. The results obtained in this experiment demonstrated that four weeks of consumption of this Bifidobacterium strain had no adverse effects on rat's general health status, blood biochemical parameters or histology. Therefore, it is likely to be safe for human use. Fecal harmful enzymes such as $\beta-glucosidase,\;\beta-glucuronidase$, tryptophanase and urease, were effectively inhibited during the administration of the B. longum SPM1205. These results suggested that this B. longum SPM 1205 could be used for humans as a probiotic strain.

• 한국균학회지
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• 제38권1호
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• pp.21-24
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• 2010

현재 시판되고 있는 세포벽 분해 효소 중 cellulase onozuka R-10(Yakult Honsha, Japan), yatalase(Takara), $Glucanex^R$ 200G(Novo Industry, Denmark)를 조합하여 Flammulina velutipes를 대상으로 가장 효율적인 방법을 선발하고, 선발된 방법을 Pleurotus ostreatus, P. eryngii, Hypsizygus marmoreus에 적용하였다. F. velutipes의 포자, 셀로판지에 배양한 균사, homogenizer로 마쇄한 균사체를 대상으로 공시한 세포벽 분해효소를 단독 혹은 조합하여 처리하였다. 그 결과, 포자현탁액이나 셀로판지에 배양한 균사를 사용하는 것에 비해 homogenizer로 마쇄한 균사체를 사용하는 것이 높은 수율로 원형질체를 분리할 수 있었다. 또한 시험에 사용한 효소 중선발된 $Glucanex^R$ 200G와 cellulase onozuka R-10 효소의 혼합 처리에서 고효율의 원형질체가 분리되었다. 분리된 원형질체를 대상으로 재생률을 조사한 결과, 0.39~0.51% 범위의 재생률을 나타내었다