본 실험에서는 외인성 효소 첨가제 및 혼합 세균 배양을 통한 고상발효(Solid-state fermentation, SSF)가 채종박(Rapeseed meal, RSM)의 체외건물소화율(In vitro dry matter digestibility, IVDMD) 및 단쇄지방산(Short chain fatty acid) 생성에 미치는 영향을 조사하기 위해 수행되었다. 외인성 효소 칵테일(첨가 및 미첨가) 및 RSM에 대한 SSF(발효 및 비발효)를 나타내는 2 x 2 요인 설계가 적용되었다. 3-step 돼지 소화율 모델을 적용하여 채종박의 건물 소화율을 분석하였으며, 72시간 대장발효 후 상층액을 수집하여 단쇄지방산 생성량을 분석한 후 칼로리 단위로 변환하여 가소화에너지 소화율을 분석하였다. 소장 (IVDMDh) 및 전장 (IVDMDt) 건물소화율에서는 고상발효된 채종박이 더 높게 나타났다 (각각 p < 0.01). 마찬가지로, 외인성 효소 첨가제 처리구에서 채종박의 소장 소화율(IVDMDh)이 증가하는 경향을 나타냈다(p = 0.06). Acetic acid 및 butyric acid의 생산은 대조구에 비해 고상발효 처리 시 유의하게 더 생산되었으며 (각각 p < 0.01), 이는 총 단쇄지방산의 생산 증가 경향을 나타냈다(p = 0.09). 에너지 소화율에서는 채종박의 고상발효 및 외인성 효소제 첨가가 유의적으로 높게 나타났다 ( p < 0.01). 그러므로 채종박의 고상발효 처리는 단백질 이용성을 비롯한 영양적 가치를 향상시키는데 효과적이라고 사료된다.

Galacto-oligosaccharides (GOS) are prebiotics that have a beneficial effect on human health by promoting the growth of probiotic bacteria in the gut, in addition to having various applications in the food industry. GOS are generally produced from lactose in a reaction catalyzed by ß-galactosidase. Synthesis of GOS from whey permeate (WP) (ultrafiltration of whey, concentrated then spray dried) using surface engineered β-galactosidase in Pichia pastoris (P. pastoris) is a novel method to convert waste into a valuable product. Cell-surface display is the expression of peptides and proteins on the surface of living cells by fusing them to functional components of cells. Surface engineered cells have many potential uses. The Flo1p flocculation functional domain, thought to be located near the N terminus, recognizes and adheres non-covalently to cell-wall components such as α-mannan carbohydrates, causing reversible aggregation of cells into flocs.

This research confirmed the diversity and characterization of halophilic microorganisms isolated from the various solar salterns, collected on the inside and outside of the country. To isolate strains, the marine agar medium was basically used and cultivated at 37oC for several days aerobically. After single colony isolation, a totally of 230 pure colonies were isolated and phylogenetic analysis was carried out based on the result of 16S rDNA sequencing, indicating that isolated strains were divided into 4 phyla, 12 families, 27 genera and 64 species. Firmicutes phylum, the main phyletic group, comprised 89.6% with 3 families, 17 genera and 52 species of Bacillaceae, Staphylococcaceae and Carnobacteriaceae. To confirm whether isolated strain can produce industrially useful enzyme or not, amylase, lipase, and protease enzyme assays were performed individually, showing that 177 strains possessed at least one enzyme activity. Especially 17 strains showed all enzyme activity tested. This result indicated that isolated strains have shown the possibility of the industrial application. Therefore, this study has contributed to securing domestic genetic resources and the expansion of scientific knowledge of the halophilic microorganisms community in solar salterns.

The aim of this study was to select various fungal strains indigenous to Korea that have the potential to produce cellulases, including filter paper activity (FPase), endo-β-1,4-glucanase (EG), and β-glucosidase (BGL). Among the 25 species of Ascomycetes and the 32 species of Basidiomycetes tested in this study, the Bjerkandera adusta KUC10565, Heterobasidion orientale KUC10556, Hyphoderma praetermissum KUC10609, and Trichoderma harzianum KUC1716 all exhibited remarkably high FPase activity. In addition, the T. harzianum KUC1716 showed high levels of EG and BGL activity. This strain has been selected for further study because of their enzymatic potential.

To evaluate effects of ligninolytic enzyme type on the mycelial response and ligninolytic enzyme production during interspecific interactions among wood-rotting fungi, 4 fungal strains, Trichophyton rubrum LKY-7, Trichophyton rubrum LSK-27, Pycnoporus cinnabarinus, and Trichoderma viride, were selected. Regarding ligninolytic enzyme production, LKY-7 secreted laccase and manganese peroxidase (MnP), P. cinnabarinus secreted only laccase, and LSK-27 secreted only MnP in glucosepeptone medium, while T. viride did not produce any ligninolytic enzymes. In the co-culture of LKY-7 with P. cinnabarinus, the formation of aerial mycelium was observed and the enhancement of laccase activity owing to interspecific interaction appeared to be very low. In the co-culture of LKY-7 and P. cinnabarinus with LSK-27, a hypha-free clear zone was observed, which resulted in deadlock, and increased laccase or MnP activity was detected at the interaction zone. The interaction responses of LKY-7, P. cinnabarinus, and LSK-27 with T. viride were characterized by the formation of mycelial barrages along the interface. As mycelial barrages were observed at the T. viride territory and no brownish pigment was observed in the mycelial barrages, it is suggested that laccase and MnP are released as part of an offensive response, not as a defensive response. The co-culture of P. cinnabarinus with T. viride lead to the highest enhancement in laccase activity, yielding more than 14-fold increase in laccase activity with respect to the mono-culture of P. cinnabarinus. MnP activities secreted by LKY-7 or LSK-27 was generally low in interspecific interactions.

The entomopathogenic fungus Metarhizium anisopliae is one of potent biological control agents against a variety ofinsect pests. In this study, we investigated enzyme production of M. anisopliae strains A and B. They produced extracellularenzymes for degrading the epidermis of Monochamus alternatus, a crucial mediator of the pinewood nematode Bursaphelenchusxylophilus. With Q-TOF MS/MS analysis, 29 kDa protein, a major band on a SDS-PAGE gel, was identified as subtilisin-likeserine protease PR1A. M. anisopliae A produced an extracellular enzyme more efficiently than M. anisopliae B: however,enzyme activities targeted for the cuticle were comparable. Our results suggest that the two strains of M. anisopliae havethe biological potential against M. alternatus with insecticidal protease production.

국내 과채류 재배단지의 뿌리혹선충 발생에 관한 조사를 위해 1997년부터 1999년까지 경북 성주군을 중심으로 경기 여주군, 경남 함안군, 충북 청원군 등에서 과채류재배지의 토양을 채집하여 식물기생선충 종류와 밀도 조사, 뿌리혹선충 암컷의 효소표현형에 의한 종 동정을 실시하였다. 경북 성주군의 185개 참외재배 포장 중 99개 포장(53.5%)에서 뿌리혹선충이 검출되었고 나선선충류(Helocotylenchus spp.)는 7개, 둥근꼬리선충류(Aphelechus spp.)는 43개, 환선충류(Criconematid)는 26개 포장에서 검출되었다. 뿌리혹선충 암컷의 Malate dehydrogenase 및 Esterase 등 2가지 효소표현형을 이용하여 한국에 분포하는 주요 4종의 동정이 가능하였다. 효소 표현형을 이용하여 성주군 선남면에서 채집된 13개 시료 중 당콩뿌리혹선충으로 동정된 것이 6포장, 고구마뿌리혹선충 5포장이었으며 2개 포장은 두 종의 혼재하는 것으로 나타났다. 성주군 초전면의 6개 포장 시료 중 4개가 땅콩뿌리혹선충, 1개가 고구마뿌리혹선충으로 동정되었으며 1포장의 뿌리혹선충은 효소표현형이 미동정 종으로 나타났다. 경기도 여주군의 참외재배단지에서는 14개 조사대상 중 당근뿌리혹선충이 11개 포장, 땅콩뿌리혹선충이 3개포장으로 당근뿌리혹선충이 우점종인 것으로 나타났다.

가용성 전분으로부터 CGTase 효소에 의한 Cyclodextrin(CD) 동족체의 생산 실험을 dead-end형 막모듈을 설치한 막-효소 반응기에서 수행하였다. 회분 반응기에서의 CD 동족체 생산시 생산물 저해작용 및 생산된 CD의 다른 환원당으로의 분해에 의해 반응시간에 따른 전분의 CD 동족체로의 총 전환율이 최대 45%를 나타내었다가 급격히 감소하였다. 반면 분획분자량 10,000 달톤의 분리막이 설치된 막-효소 반응기에서의 CD 동족체 생산시 CD 동족체를 생산 즉시 반응기로부터 막을 통해 연속 분리할 수 있어 총 전환율이 35%로 일정하게 유지되었다. 10% 전분용액 및 2 atm 조작압력하에서 막-효소 반응기를 24시간 운전하였을 때 CD 동족체의 누적 생산량은 약 3.7 kg/m2이었다.

본 연구에서는 저온추출 뿐만 아니라 효소처리를 이용하여 생과형 블루베리 과일음료생산을 위한 최적의 방법을 확립하고자 하였다. 생리학적 기능성물질 추출 시 열에 의한 영양손실을 막기 위해 저온을 사용하였다. 또한 다양한 효소처리를 이용하여 혼탁 방지 및 추출수율 향상을 위한 최적의 추출조건을 확립하고자 하였다. 블루베리를 cellulase, pectinase 및 cellulase:pectinase(1:1) 혼합 효소와 효소 처리량, 추출온도, 추출시간, 추출 교반속도 등을 고려한 다양한 조건으로 추출하였을 때, 가장 우수한 추출 조건을 조사하였다. cellulase로 처리할 경우, 추출률이 85.72-86.55%, pectinase 처리구는 87.06-87.93%, cellulase:pectinase(1:1) 혼합 처리할 경우, 86.84-88.14%의 추출률을 나타냈다. 추출 온도는 45℃에서 87.91±0.05%, 3시간 추출 하였을 경우 87.88±0.10%, 교반속도는 90 rpm에서 가장 높은 추출결과를 보였다. 블루베리 추출액의 당도 및 pH는 추출 조건에 관계없이 모든 처리구에서 통계학적으로 유사한 결과를 보였으나, 총 폴리페놀 함량은 pectinase 처리구에서 18.62 mg/g으로 가장 높은 값을 나타냈다. 유리당 함량은 모든 시료에서 과당과 포도당 두 가지 당류만 검출되었으며 cellulase 효소 0.10%를 처리한 블루베리 추출액의 유리당 함량이 포도당 0.51%, 과당 0.49%로 가장 높았으나 다른 처리구와 유의적인 차이는 없었다. 블루베리를 이용하여 생과일형 음료 제조를 위한 최적 조건은 cellulase: pectinase(1:1) 혼합효소 0.1%를 첨가하여 45℃ 온도에서 90 rpm의 교반속도로 조사되었다.

Since biodiesel as bioenergy is defined as ester compounds formed by esterification of animal/vegetable oils, in this study three vegetable cooking oils (market, waste and refined waste ones) were esterified by reactions of alkali catalyst and immobilized enzyme. The fatty acid composition of the formed ester compounds was analyzed to investigate the feasibility of biodiesel production. By lipolysis (i.e, hydrolysis of Triglyceride (TG)), all three vegetable oils used in this study were found to produce Diglyceride (DG), Monoglyceride (MD) and Fatty acid ethylester (FAEE). However, the amount of produced FAEE (which can be used as an energy source) was in the increasing order of market cooking oil, waste one and refined waste one. With NaOH catalyst, FAEE was produced about 24.92, 17.63 and 11.31 % for the respective oils while adding Lipozyme TL produced FAEE about 43.54, 38.16 and 24.47 %, respectively. This indicates that enzyme catalyst is more effective than alkali one for transesterification. In addition, it was found that the composition of fatty acids produced by hydrolysis of TG was unchanged with alkali and immobilized enzyme reactions. Thus it can be expected that stable conditions remain in the course of mixing with gasoline whose composition is similar to that of the fatty acids.

전통발효식품인 된장으로부터 높은 활성의 fibrinolytic enzyme를 생산하는 미생물을 수십 종 분리하였다. 산업적으로 우수한 균종을 선별하기 위하여 분리된 미생물중 fibrinolytic enzyme활성과 성장 속도면에서 가장 우수한 균주를 선별하여 하였으며, 형태학적, 생화학적 및 생리학적 특성을 조사한 후 Bacillus sp.로 동정되었다. Fibrinolytic enzyme생산을 위한 최적 배양조건은 초기 pH 일 때 상대활성도가

Total RNAs were isolated from cultured roots of Scopolia parviflora, poly(A)+ RNA was obtained through the mRNA purification, cDNA library of Hnh6h was constructed. Recombinant baculoviruses in Spodoptera frugiperda (Sf) cells were constructed by use of the transfer vector pBacPAK, which has the AcNPV sequence under the polyhedrin promoter. The expression vector carrying Hnh6h gene was transferred to S. parviflora and obtained transgenic hairy root lines. Our results confirmed the over expression of the H6H protein was used by anti-pBacPAK about cDNAs of S. parviflora. This study will served for production of tropane alkaloids by metabolic engineering.