• Archives of Pharmacal Research
• /
• 제20권3호
• /
• pp.206-211
• /
• 1997

The optimal conditions for the production and regeneration of the protoplasts from Lentinula edodes were studied. Protoplast formation from the mycelia of L. edodes which were cultured in liquid medium showed a significantly high yield compared with that of the mycelia which were cultured on cellophane covered agar media. A mixture of Novozyme 234 (15 mg/ml) and Cellulase Onozuka R10 (10 mg/ml) in 0.6 M mannitol (pH 4) was optimal lytic enzyme for the protoplast release. The optimal incubation time and mycelia age were 3.5-4 hours at $30^{\circ}C$ and 6-8 days, respectively. Regeneration frequency was 0.18% plated onto a medium containing 0.6 M sucrose, and 0.08% plated onto a medium containing mannitol. But hardly any regeneration was observed in the media containing NaCl, KCl, or $MgSO_{4}$ More than 90% of the protoplasts contained nuclei and the nucleus number per protoplast was 1.1. The DNA content per nucleus was 5.1 pg. The diameter of the protoplast was $3-5{\mu}m$ and it had a well defined cell structure.

• Journal of the Korean Wood Science and Technology
• /
• 제21권3호
• /
• pp.53-60
• /
• 1993

Substrates used were hardwood-Suwon poplar-(Populus alba${\times}$glandulosa L.) and softwood-pitch pine-(Pinus rigida M.). And these substrates were steam exploded then treated with sodium chlorite at 75$^{\circ}C$ with occasional stirring in order to obtain samples which had different lignin contents and crystallinity. And then this resulting samples incubated with a commercial cellulase derived from Trichoderma ressei. The contents of Klason lignin were decreased as the increasing of the ratio of sodium chlorite in the two species. The effect of hardwood was more effective than that of softwood in the same ratio of sodium chlorite. The minimum contents of Klason lignin were 0.8% and 5.1% respectively. And the crystallinities of cellulose were increased very little as increasing of the ratio of sodium clorite. The hydrolysis extent of the two species were increased as the increasing of delignification. Especially, the hydrolysis extent of hardwood was more higher than that of softwood. The maximum hydrolysis extent were 89.8% and 71.1%, respectively.

• Journal of Applied Biological Chemistry
• /
• 제61권1호
• /
• pp.59-67
• /
• 2018

본 연구는 경상남도 산청군 소재 경남과학기술대학교 학술림에서 채취한 토양으로부터 CMCase와 xylanase를 생산하는 3개의 Bacillus종을 분리하였다. API kit 분석과 16S rRNA 유전자 염기서열 분석을 통해 3개의 균 모두 Bacillus종에 속하였으며, Bacillus sp. EFL1, EFL2, EFP3로 명명하였다. Bacillus sp. EFL1, EFL2, EFP3의 최적 생장온도는 $37^{\circ}C$였으며, CMCase와 xylanase의 활성은 배양 후 12시간에 최고에 달하였다. Bacillus sp. EFL1의 CMCase 효소활성의 최적온도는 $50^{\circ}C$, pH는 5.0이었고, xylanase 효소활성의 최적온도는 $50^{\circ}C$, pH 6.0이었다. Bacillus sp. EFL2의 CMCase활성의 최적온도는 $60^{\circ}C$, pH는 5.0이었고, xylanase 효소활성의 최적온도는 $60^{\circ}C$였고, pH 3.0-9.0까지 비교적 높은 활성을 보였다. Bacillus sp. EFP3의 CMCase의 최적온도는 $50^{\circ}C$, pH는 5.0이었고, xylanase의 최적온도는 $50^{\circ}C$, pH 4.0이었다. Bacillus sp. EFL1, EFL2, EFP3의 CMCase는 모두 온도안정성이 낮았다. 또한 Bacillus sp. EFL1과 EFP3의 xylanase 역시 온도안정성이 낮았지만, Bacillus sp. EFL2의 xylanase는 온도안정성이 높았다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제16권11호
• /
• pp.1610-1613
• /
• 2003

The aim of the experiment was to study the changes in the activities of various rumen fibre degrading enzymes due to the feeding of chemically treated mustard (Brassica campestris) straw in sheep. Mustard straw (MS) (<5 cm particle size) was treated either with urea (4% (w/w), or with 2% sodium hydroxide (NaOH), or with alkaline hydrogen peroxide (2% NaOH and 1.5% hydrogen peroxide ($H_2O_2$)) and/or supplemented with 2% (w/w) urea. Seven maintenance type rations were prepared using MS (70 parts) with molasses (5 parts) and concentrate (25 parts). They were untreated MS (CMS), urea treated MS (UMS), urea supplemented MS (MSUS), alkali treated MS (AMS), alkali treated and urea supplemented MS (AMS-US), alkali $H_2O_2$ treated MS (AHMS) and alkali $H_2O_2$ treated and urea supplemented MS (AHMS-US). They were then compressed into a complete feed block with the help of block making machine. Forty two male hoggets of Malpura breed sheep were equally distributed into each treatment group and (were) offered feed and water ad libitum. At the end of 21 days of feeding trial, rumen liquor was collected through stomach tube from three animals in each group at 0 h, 4 h, 8 h, 12 h of post feeding. Results showed that the level of enzyme varied from 8.52 to 11.12, 40.85 to 50.37, 3.22 to 3.78, 2.09 to 2.77 and 31.44 to 44.24 units/100 ml SRL respectively for carboxymethyl cellulase (CMCase), $\alpha$-amylase, microcrystalline cellulase (MCCase), filter paper (FP) degrading enzyme and $\alpha$-glucosidase. Processing of MS affected the enzyme activities, in a way, that NaOH and AHP treatment significantly reduced CMCase and FP degrading enzyme. The effect of urea treatment showed an increase in the activity of MCCase and $\alpha$-glucosidase. But the supplementation of urea increased the activity of CMCase, FP degrading enzyme and $\alpha$-glucosidase. The CMCase, $\alpha$-amylase, $\alpha$-glucosidase activities were highest at 4hr whereas MCCase and FP degrading enzyme had maximum activities at 12 h post feeding Results suggested that MS might need longer time in the rumen for its effective degradation.

• 한국연초학회지
• /
• 제13권2호
• /
• pp.5-20
• /
• 1991

For the quality enhancement of harvested-year leaf tobacco to the quality of 2-year naturally aged leaf tobacco, cellulose and nicotine degradative bacteria were isolated and identified. Effects of artificial fermentation treated cellulase and nicotine degradative bacteria on the quality of leaf tobacco were investigated from the chemical and sensory points of view. 1, Changes in chemical composition of leaf tobacco resulted from the addition of cellulase extracted from Cellulomonas sp. [3ml(${\mu}{\textrm}{m}$ D-glucose/ml. mil-1) of enzymes solution 11009 of leaf tobacco] and nicotine degradative bacteria, Pseudomonas sp. 2ml(IX109 cells$\div$ 100g of leaf tobacco), and subsequently fermented at 40${\mu}{\textrm}{m}$$^{\circ}C$, 65% R. H. for 40 days are as follows : 1) Content of crude fiber decreased 12% It took 9 min, 53 sec. to reach full combustion in control group but took only 7 min. 47 sec. in the treated group, taking almost equal time to 2-year naturally aged leaf tobacco(7 min. 35sec.). 2) Light intensity of control group was 60.96% with bright lemon color but that of treated leaf tobacco accounted for 47.69 with orange to dark brown color series, which was almost equal to the value, 45.69, of 2-year naturally aged leaf tobacco. 3) Linoleic acid, serving mild taste among organic acids, amounted to 1.llmg/g in control group but increased to 1.35m9/9 in the treated leaf tobacco, identical to the content(1.35mg/g) of 2-year naturally aged leaf tobacco. 4) Content of solanone, on of the typical leaf tobacco flavor compounds, accounted for 2.95% in control group but increased to 2.87% in treated group. 5) Methyl furan, useful flavor compound in smoke composition, accounted for 17.6$\mu\textrm{g}$/cig. in control group but increased to 25.9$\mu\textrm{g}$/cig. in treated group. However, acroleine decreased from 69.3$\mu\textrm{g}$/cig. in control group to 58.6$\mu\textrm{g}$/cig. in treated group 2. In sonsory test, mild taste evaluation of control group scored 5.47 and treated group 7.93 which was evaluted almost equal to the value(8.00) of 2-year naturally aged leaf tobacco. Aroma evaluation of control group scored 5.60, treated group 8.20, and 2-year naturally aged leaf tobacco 8.33. In addition, total harmony taste of control group showed 5.67, treated group 8.07 (p<0.01), and 2-year naturally aged leaf tobacco 8.00. From these results, it can be said that quality of treated leaf tobacco is not inferior to that 2-year naturally aged leaf tobacco.

• Journal of Microbiology and Biotechnology
• /
• 제16권8호
• /
• pp.1255-1261
• /
• 2006

An alkaliphilic bacterium, Bacillus sp. strain BK, was found to produce extracellular cellulase-free xylanolytic enzymes with xylan-binding activity. Since the pellet-bound xylanase is eluted with 2% TEA from the pellet of the culture, they contain a xylan-binding region that is stronger than the xylan-binding xylanase of the extracellular enzyme. The xylanases had a different molecular weight and xylan-binding ability. The enzyme activity of xylanase in the extracellular fraction was 6 times higher than in the pellet-bound enzyme. Among the enzymes, xylanase had the highest enzyme activity. When Bacillus sp. strain BK was grown in pH 10.5 alkaline medium containing xylan as the sole carbon source, the bacterium produced xylanase, arabinofuranosidase, acetyl esterase, and $\beta$-xylosidase with specific activities of 1.23, 0.11, 0.06, and 0.04 unit per mg of protein, respectively. However, there was no cellulase activity detected in the crude enzyme preparation. The hydrolysis of agricultural residues and kraft pulps by the xylanolytic enzymes was examined at 50$^{\circ}C$ and pH 7.0. The rate of xylan hydrolysis in com hull was higher than those of sugarcane bagasse, rice straw, com cop, rice husk, and rice bran. In contrast, the rate of xylan hydrolysis in sugarcane pulp was 2.01 and 3.52 times higher than those of eucalyptus and pine pulp, respectively. In conclusion, this enzyme can be used to hydrolyze xylan in agricultural residues and kraft pulps to breach and regenerate paper from recycled environmental resources.

• Applied Biological Chemistry
• /
• 제23권1호
• /
• pp.64-72
• /
• 1980

본 연구에서는 산업폐수중 유기성분이 많은 식품, 펄프 공장폐수에 대하여 유기들을 영양원으로 하는 미생물을 배양시켜 유기성분을 경감시켜 폐수 정화를 목격으로 하였다. 한편 폐수정화에서 회수되는 균체는 단백질자원으로서의 사료화를 또 다른 목적으로 하고 일련의 실험을 통하여 다음과 같은 결과를 얻었다. 1. 펄프, 맥주, 빵효모, 주정공장폐수의 총당은 $1.4{\sim}1.54%$, 총 질소는 $0.25{\sim}0.35%$, BOD는 $400{\sim}25,000$, COD는 $500{\sim}28,000$, pH는 $3.8{\sim}7.0$이었고 주정공장 폐수가 BOD, COD는 가장 높았다. 2. 각 폐수중 세균총수는 $4{\times}10^4{\sim}1{\times}10^9$, 효모총수는 $2{\times}10^2{\sim}7{\times}10^4$이었다. 3. 폐수별로가 생장속도 우수한 균주로는 펄프공장폐수에서는 세균을, 맥주, 빵효모, 주정공장폐수에서는 효모를 선정 하였다. 4. 주정공장폐수정화에 최적인균주로서는 Candida curvata SAFM 70, Saccharomyces cerevisiae SAFM 1008을 선정하였다. 5. 주정중류폐액 및 중류잔사에 미생물성 cellulase, xylanase, pectinase를 처리함으로서 처리전보다 잔사량을 36% 감소시키면서 가용성화를 높여서 환원당량을 1.3배 증가시켰으므로 효모배양에 좋은 조건임을 확인하였다. 6. 효소처리한 주정증류제액에 선정된 효모를 배양하면서 배양최적조건을 확인하였다. a. Candida curvata SAFM 70 및 Saccharomyces cerevisiae SAFM 1008 2균주 모두 최적 pH는 5.0이었다. b. 2균주 모두 질소원으로는 요소, 질소농도로는 0.2%가 최적조건이었고 인삼염으로서는 $KH_2PO_4$, 그의 농도는 0,1% 그러고 Mg원으로서는 $MgSO_4$, 그의 농도는 0.02% 이었다. c. 2균주 모두 최적온도는 $30^{\circ}C$이었고 통기, 교반효과는 증식도에 크게 영향끼침을 알었다. 7. 종효모의 생포수는 적어도 $1.8{\times}f10^5/ml$ 이상이어야 증식이 순조로움을 알었다. 8. 미생들 효소를 처리한 주정증류폐액에 선정효모를 배양하여 건조균체를 생산하는데는 Saccharomyces cerevisiae SAFM 1008 보다 Candida curvata SAFM 70이 우수하였다. 9. 미생물성효소를 처리한 주정증류폐액 1000ml 당 Saccharomyces cerevisiae SAFM 1008은 16g, Candida curvata SAFM 70은 17.6g의 건조균체를 해수할 수 있었다. 주정증류공장폐수의 BOD 제거율은 전자의 효모에서 46a, 후자의 효모에서 52%이었다. 10. 주정증류공장폐수에서 생산된 효모균체는 단백질 함량이 $46{\sim}$52%임으로 사료용효모로서 적당함을 확인하였다.

• 농업과학연구
• /
• 제4권2호
• /
• pp.283-316
• /
• 1977

한국(韓國)에 있어서 벼 잎집무늬마름병(病)의 발생변동(發生變動), 병원균(病原菌)의 배양적(培養的) 특성(特性)과 병원성(病原性), 발병환경(發病環境), 재배법(栽培法)과 발병관계(發病關係), 품종(品種)의 저항성(抵抗性) 등(等)에 관(關)하여 연구(硏究)한 결과(結果)를 요약(要約)하면 다음과 같다. 1. 한국(韓國)에 있어서 벼 잎집무늬마름병(病)의 발생면적(發生面積)은 다비재배(多肥栽培), 조기조식재배(早期早植栽培) 및 통일계품종(統一系品種)의 확대보급등(擴大普及等)으로 인(因)하여 매년(每年) 증가(增加)되었고 특(特)히 1971년(年)부터 전국적(全國的)으로 급격(急激)히 발생면적(發生面積)이 증가(增加)되어서 1976년(年)에는 전체수도재배면적(全體水稻栽培面積)의 약 65.2%에서 본병(本病)이 발생(發生)하였다. 2. 우리나라에 분포(分布)하는 벼 잎집무늬마름병균(病菌)의 배양적(培養的) 성질(性質) 및 병원성(病原性)에는 많은 변이(變異)가 있었으며 병원균(病原菌)의 생육최적(生育最適) 온도(溫度)도 $25^{\circ}C$에서 $30^{\circ}C$사이로 균주간(菌株間)에 차이(差異)가 있었다. 한편 병원균(病原菌)의 배양적(培養的) 성질(性質)과 병원성(病原性)에는 상관관계(相關關係)가 없었다. 3. 잎집무늬마름병균(病菌)의 PDA배지상(培地上)에서의 균사생장(菌絲生長)은 채집지(採集地)의 기온(氣溫)과 밀접(密接)한 관계(關係)가 있었다. 즉 채집지(採集地)의 기온(氣溫)이 높은 지역(地域)의 균주(菌株)들은 기온(氣溫)이 낮은 지역(地域)의 균주(菌株)들에 비(比)해 $35^{\circ}C$ PDA배지(培地)에서 균사생장(菌絲生長)이 양호(良好)하였고 반대로 $12^{\circ}C$ PDA배지상(培地上)에서는 기온(氣溫)이 낮은 지역(地域)에서 채집(採集)한 균주(菌株)들이 기온(氣溫)이 높은 지역(地域)에서 채집(採集)한 균주(菌株)들 보다 균사생장(菌絲生長)이 양호(良好)하였다. 4. 잎집무늬마름병균(病菌)을 접종(接種)한 수도엽(水稻葉)으로부터 얻은 Pectin polygalacturonase 및 Cellulase의 역가(力價)는 접종(接種)3일(日)째에 최대(最大)였고 Pectin methylestrase의 역가(力價)는 접종(接種) 4일(日)째에 최대(最大)였으며 병원성(病原性)이 강(强)한 균주(菌株)에서는 Pectin methylestrase의 활성(活性)이 높은 경향(傾向)이었고 병원성(病原性)이 약한 균주(菌株)에서는 Pectin methylestrase의 활성(活性)이 낮은 경향(傾向)이었으나 다른 효소(酵素)들은 병원성(病原性)과 관계(關係)가 없었다. 5. 배지(培地)에 형성(形成)된 균핵(菌核)을 건조처리(乾燥處理)하였을 경우(境遇)의 저온저항성(低溫抵抗性)은 매우 강(强)하여 $-20^{\circ}C$에서도 발아력(發芽力)을 상실(喪失)치 않았으나 습윤처리(濕潤處理) 하였을 경우(境遇)에는 저항성(抵抗性)이 매우 약(弱)하여 $-5^{\circ}C$에서도 발아력(發芽力)을 상실(喪失)하였다. 포장(圃場)에서 월동(越冬)한 균핵(菌核)의 발아율(發芽率)도 답토양(沓土壤)의 건습(乾濕)에 따라 18%~70%의 차이(差異)를 나타내며 담수상태(湛水狀態)에서 월동(越冬)한 균핵(菌核)은 건조(乾燥)한 토양(土壤)에서 월동(越冬)한 균핵(菌核)보다 발아율(發芽率)이 낮았다. 6. 잡초(雜草)에 대(對)한 기생성(寄生性)을 조사(調査)하였던 바 벗풀(Sagittaria trifolia L.), 물달개비(Monochoria vaginalis Presl), 여뀌(Polygonum hydropiper L.). 피(Echinochloa crusgalli P. Beauv), 한련초(Eclipta prostrata L.), 강아지풀(Septaria viridis P. Beauv.), 좀바랭이(Digitaria sangvinalis Scapoli) 등(等)에서 잎집무늬마름병반(病斑)이 형성(形成)되었다. 7. 질소배비구(窒素倍肥區)에서는 잎집무늬마름병(病)의 피해(被害)가 증가(增加)하였고 가리배비구(加里倍肥區)에서는 감소(減少)하였으나 규산시용구(硅酸施用區)에서는 피해감소(被害減少)의 효과(效果)가 없었다. 규산시용구(硅酸施用區)에서는 도체내(稻體內)의 질소함량(窒素含量)과 전분함량(澱粉含量)이 다같이 감소(減少)하였다. 8. 생육전기(生育前期)의 이병경율(罹病莖率)은 밀식구(密植區)가 높았으나 생육후기(生育後期)에는 소식구(疎植區)의 1주당(株當) 분얼수(分蘖數)가 많아 이병경율(罹病莖率) 및 피해도(被害度)가 높아졌으며 주당(株當) 1본식(本植)한 구(區)에서는 피해(被害)가 적었다. 9. 잎집무늬마름병(病)에 대(對)한 수도(水稻) 품종(品種)의 피해도차이(被害度差異)는 출수일수(出穗日數)와 밀접(密接)한 관계(關係)가 있었다. 즉 출수일수(出穗日數)와 피해도(被害度)와의 상관관계(相關關係)는 6월(月)13일(日) 이앙(移秧)한 구(區)에서는 r=-0.707이었고 6월(月) 25일(日) 이앙(移秧)한 구(區)에서는 r=-0.496으로 비교적 높은 상관(相關)을 나타내어 출수(出穗)가 빠른 것이 피해(被害)가 크고 낮은 것이 피해(被害)가 적은 경향(傾向)이었다. 또한 본병(本病)은 분얼(分蘖)이 많은 품종(品種)일수록 피해(被害)가 심(甚)하였다. 10. 생육전기(生育前期)의 이병경율(罹病莖率)은 출수후(出穗後)의 피해도(被害度)와 상관(相關)이 없었으나 생육후기(生育後期)의 이병경율(罹病莖率)은 피해도(被害度)와 상관(相關)이 높았다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제12권1호
• /
• pp.28-35
• /
• 1999

The effects of addition of celulases (A cremonium cellulolyticus and Trichoderma viride, CE), a commercial inoculum containing lactic acid bacteria (Lactobacillus casei, LAB), fermented green juice (macerated napier grass with water was incubated anaerobically with 2% glucose for 1 day, FGJ) and glucose (G), and regional difference of ensiling on napier grass (Pennisetum purpureum Schum.) silage were studied by using 900 ml laboratory glass bottle silos under 30 and $40^{\circ}C$ storage conditions in 1995 and 1996. Experiment 1 was carried out to compare the addition of CE, LAB, FGJ and the combinations. Silages were stored for 45 days after ensiling. Experiment 2 studied the effects of applications of CE, LAB, FGJ and G. Experiment 3 was carried out using the similar additives as experiment 2 except for LAB. Silages were stored for 60 days in the experiments 2 and 3. Experiments 1 and 2 were done in Nagoya, and experiment 3 in Okinawa. Sugar addition through CE or G improved the fermentation quality in all the experiments, which resulted in a greater decrease in the pH value and an increased level of lactic acid, while butyric acid contents increased under $30^{\circ}C$ storage condition in CE addition. LAB and FGJ additions hardly affected the silage fermentation quality without additional fermentable carbohydrate. But the combination of LAB, FGJ and glucidic addition (CE and G) improved the fermentation quality. The effect of the regional difference of ensiling between temperate (Nagoya; $35^{\circ}$ N) and subtropical (Okinawa; $26.5^{\circ}$ N) zones on silage fermentation quality was not shown in the present study.

• 한국균학회지
• /
• 제43권3호
• /
• pp.165-173
• /
• 2015

감자덩이버섯(Mattirolomyces terfezioides)은 단맛을 가진 트러플의 한 종류로, 주로 아까시나무(Robinia pseudoacacia)와 외생균근을 형성한다. 상업적으로 인공재배할 만한 가치가 있는 버섯으로, 본 연구에서는 우리나라 아까시나무 숲에서 수집한 감자덩이버섯균의 균사 생장특성과 효소활성에 대해 조사한 바를 처음으로 보고하고자 한다. 이 균은 PDA 배지, $30^{\circ}C$에서 가장 잘 생장하였고, 액체배양 시에는 pH 8.0의 MMN 배지에서 균 생장력이 가장 우수하였다. 또한 암모늄염보다는 질산염 형태의 무기질소원이 함유된 액체배지에서 균체량이 1.8배 더 높았으며, 효소 검색용 고체배지 상에서 carboxymethylcellase와 laccase 활성을 나타냈다.