• 한국균학회지
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• 제32권2호
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• pp.125-129
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• 2004

섬유소분해효소(CMCase)를 산업적으로 이용하기 위한 기초자료를 제공하고자 섬유소분해능이 우수한 L. japonicus로부터 CMCase를 분리 정제하였다. L. japonicus의 배양액으로부터 4단계를 거쳐 분자량이 42 kDa인 CMCase를 분리 정제하였다. 이 효소는 pH 6.0에서 최적의 활성을 보여주는 acidic CMCase로서 $30^{\circ}C$에서 최대 활성을 나타냈다. EDTA에 의해 활성이 저해되는 것으로 보아 metalloenzyme으로 추정되며, SDS에 의해 저해되는 것으로 보아 S-S기를 갖고 있는 효소로 판단된다. $Al_{2}(SO_{4})_{3}$와 $BaCl_{2}$에서는 효소 활성이 높았으나 그 이외의 금속염에서는 효소 활성이 낮았다.

• 한국균학회지
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• 제33권2호
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• pp.75-80
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• 2005

섬유소분해능이 우수한 L. roseoalbus로부터 분리 정제한 CMCase의 활용에 대한 기초적인 자료를 제공하고자 실험하여 L. roseoalbus의 배양액으로부터 4단계를 거쳐 분자량이 28.5 kDa인 CMCase를 분리 정제하였다. 이 효소는 pH 4.0에서 최적의 활성을 보여주는 acidic CMCase로 $30^{\circ}C$에서 최대 활성을 나타냈다. EDTA에 의해 활성이 저해되는 것으로 보아 metalloenzyme으로 추정되며, PMSF에 의해 저해되는 것으로 보아 serine 잔기를 갖고 있는 효소로 판단된다. $Al_{2}(SO_{4})_{3}$와 $FeSO_{4}$에서는 효소활성이 높았으나 $CaCl_{2}$와 $Na_{2}MoO_{4}$에서는 효소 활성이 낮았다.

• 한국균학회지
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• 제30권2호
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• pp.113-118
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• 2002

S. rugosoannulata의 배양액으로부터 4단계를 거쳐 분자량이 54 kDa인 CMCase를 분리 정제하였다. 이 효소는 pH 4.0에서 최대의 활성을 보여주는 acidic CMCase로 $40^{\circ}C$에서 최대활성을 나타냈다. EDTA에 의해 활성이 저해되는 것으로 보아 metalloenzyme으로 추정되며 1,10-phenanthroline과 KCN 및 L-cystein 등의 저해제에 효소활성이 크게 감소하였으며, $AgNO_{3},\;MgSO_{4},\;KCl$등의 금속염에서는 효소활성이 증가되었으나, $ZnSO_{4},\;BaCl_{2},\;CaCl_{2}$ 등에서는 효소활성이 저해되었다.

• Journal of Microbiology
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• 제34권4호
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• pp.305-310
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• 1996

A microorganism producing carboxymethyl-cellulase (CMCase) was isolated from 300 soil and compost samples. The isolate was identified as Bacillus sp. by $Biolog^{TM}$ test and fatty acid analysis, and named as Bacillus sp. KD1014. The isolate could degrade, in addition to CMC, various kinds of polysaccharides such as levan, xylan, starch, and filter paper but hardly degrade microcrystalline Avicel. The optimum growth and CMCase production of the isolate was observed between 16-and 25 hr-culture at 45$^{\circ}C$ and pH 5.0. The maximum CMCase activity was observed at pH 4.5 and 6$0^{\circ}C$. The CMCase was found to bind to Avicel. The CMCase was internally cleaved as growth continued. When crude supernatant was used for activity staining, three major bands were detected on a native gel, however, only one major band was detected on a denaturating gel after removal of the detergent.

• 미생물학회지
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• 제30권5호
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• pp.403-409
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• 1992

STA1 유전자의 promoter 와 분비신호서열을 이용하여 B. subtilis 의 CMSase 를 분비하는 재조합 효모균주를 육성하였다. STA1A 유전자의 promoter, 분비신호서열, TS region 및 mature glucoamylase N-말단부위의 아미노산 98개와 B. subtilis 의 CMCase 구조유전자가 차례로 연결된 재조합플라스미드 pYESC24 를 제작한후 효모에 형질전환하였으나 CMCase 가 세포외로 분비되지 않았다. 반면에 STA1 의 TS region 및 mature glucoamylase N-말단 아미노산 98 개를 제거하여 CMMase 구조유전자갸 STA1 의 분비신호서열에 바로 연결된 재조합 플라스미드 pYESC11 에 의한 효모형질전환 균주는 CMCase 분비능이 아주 우수하였다. 이 형질전환 균주를 YPD 배지에서 4 일간 배양한 후 세포부위 별 CMCase 역가를 측정한 결과 배양액 1 m/당 총역가 44.7 unit 존재하였으며 이중 93% 이상이 배양상등액에서 관찰되었다.

• 미생물학회지
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• 제23권2호
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• pp.147-156
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• 1985

Seasonal and vertical variations of abiontic soil carboxymethylcellulase (CMCase) activities were assessed every other month for a year in two contrasting forest soils and evaluated the relationships between soil CMCase activity and environmental parameters. In climax deciduous soil, variations in CMCase activities caused by differences in sampling time were greater than those caused by differences in soil depth. On the other hand, counter phenomenon was obserned in coniferous soil at the stage of development. Correlation analyses showed that soil CMCase activities were significantly (p>0.01) correlated with microbial respiration rates ($O_2$ uptake) and all of the microbial population sizes. From these results, it is suggested that determination of abiontic soil CMCase activity is an useful additional index for evaluating the overall microbial growth and activity in soils.

• 한국균학회지
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• 제21권1호
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• pp.28-37
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• 1993

섬유소 분해균인 P. verrculosum의 배양 여액으로부터 endo형 cellulase 인 CMCase IV를 정제하였다. CMCase IV는 13%의 탄수화물과 4.0의 pl값을 가진 산성, 당단백질이었다. CMCase IV의 SDSPAGE 상에서 분자량은 52 KDa이었으며, 효소 활성을 위한 최적 pH 및 온도는 5.0과 $50^{\circ}C$ 였다. CMCase IV를 CMC에 반응시 대부분 glucose와 cellobiose가 생산되었으며, 또한 동시에 transglycosylation 작용을 함께 갖는 것으로 사료되었다. Cellulase 활성 염색법(zymogram)을 통해서 P. verruculosum의 cellulase component가 배지 내에서 aggregation 되어있지 않음을 알 수 있었다. P. verruculosum mRNA의 in vitro 번역을 통하여 CMCase IV를 coding하는 번역산물이 동정 되었다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제3권1호
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• pp.44-47
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• 1998

Maximum cellulase production was sought by comparing the activities of the cellulases produced by different Trichoderma reesei strains and Aspergillus niger. Trichoderma reesei Rut-C30 showed higher cellulase activity than other Trichoderma reesei stains and Aspergillus niger that was isolated from soil. By optimizing the cultivation conditions during shake flask culture, higher cellulase production could be achieved. The FP(filter paper) activity of 3.7U/ml and CMCase (Carboxymethylcellulase) activity of 60U/ml were obtained from shake flask culture. When it was grown in 2.5L fermentor, where pH and DO levels are controlled, the enzyme activities were 133.35U/ml (CMCase) and 11.67U/ml(FP), respectively. Ammonium sulfate precipitation method was used to recover enzymes from fermentation broth. The dried cellulase powder showed 3074.9U/g of CMCase activity and 166.7U/g of FP activity with 83.5% CMCase recovery.

• 한국미생물·생명공학회지
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• 제25권4호
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• pp.346-353
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• 1997

During the screening of cellulase producing microorganisms, a fungal strain C-4 was selected from etiolated leaves. Based on taxonomic studies, the fungus could be classified as a strain of Trichoderma sp. When the strain C-4 was cultured in Mandels' media at 28$circ$C for 6 days, the enzyme activities detected in broth were as follows: 8.2 U/ml (28.1 U/mg) of CMCase activity, 0.75 U/ml (2.58 U/mg) of Avicelase activity, 1.67 U/ ml (5.68 U/mg) of $eta$-glucosidase activity. The optimum pH for enzyme induction was 6.2. The crude enzyme retained 100% of its original CMCase activity at 50$circ$C for 1 hr (pH 5.0), and at 4$circ$C for 24 hrs (pH 5.0). There was no effect on the CMCase activity in the presence of 1 mM of CsCl, LiCl, MgCl$_{2}$, and FeCl$_{2}$, respectively. When the crude enzyme was treated with trypsin and chymotrypsin (2% W/w) for 10 minutes, the remaining CMCase activity was 70%, but there was no further loss of activity for 60 minutes treatment at 30$circ$C. The crude enzyme showed the synergism with rumen fluid for the hydrolysis of Avicel and CMC by 118% and 130%, respectively.

• 미생물학회지
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• 제17권4호
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• pp.187-192
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• 1979

Dennis (1972) reported that Trichosporon cutaneum FRI-425 from the petioles of Pheum rhamponticum var, had showed the celluloytic activity. Chun (1977) also suggested that Trichoporon pullulons 225 isolated from the saline water of the Yeoung San River had a similar properties. However, the assay conditions for enzyme activity were not yet investigated. Thus, the present work was undertaken to examine some conditions for CMCase activity and at the same time to compare the activities of crude enzyme produce from above two species of Trichosporon pullulans. The results are as follows; 1. The maximum production of total reducing sugar by crude enzyme of Tr. pululans was after 30 minutes, whereas that of Tr. cutanuem FRI-425 was after 90 minutes. This fact showed that the reaction velocity of enzyme from Tr. pullulans 225 was more faster than that of Tr. cutaneum FRI-425. 2. Two species showed a similar trend to increase the production of reducing sugar in proportion to the increment in substrate concentration and to arrive at maximum level at lmg/ml of substrate concentration. However, Tr. pullulans 225 produced more $50{\mu}g$ of reducing sugar compared to Tr. cutaneum. 3. The optimum PH for CMCase activity is 5.0 for Tr. pullulans 225 as well as Tr. cutaneum FRI-425, and PH stability lie within the range of 6 and 8. In the activity and stability of enzyme on PH changes, enzyme of Tr. cutaneum FRI-425 was more unstable than that of TY. pullulans 225. 4. The optimum temperature for CMCase activity was $40^{\circ}C$, and enzyme activity from Tr. pullulans 225 was more sensitive to temperature changes compared with that of TY. cutaneum. The heat stability was within $40^{\circ}C$, but that was rapidly decreased above $40^{\circ}C$. In comparison of the heat stability for enzyme of Tr. cutaneum FRI-425 with that of Tr. pullulans 225 at the same temperature of $80^{\circ}C$, the former was some 10 percent more stable than the latter.