• 한국버섯학회지
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• 제16권4호
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• pp.331-333
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• 2018

The incorporation of Shiitake culture into sawdust is a widely utilized technique that can assist in reducing the cost and time consumption associated with oak cultivation. In sawdust cultivation, browning of the surface mycelia is an important stage with respect to the utility and longevity of the sawdust media. Surface browning forms a protective coating on the substrate, which can inhibit the invasion of pathogens and suppress water evaporation. Several different light sources (red LED, white LED, blue LED, and fluorescent light) were used and the intensity of illumination was carefully controlled (1.5, 10.5, $20.5{\mu}mol/m^2s$ for LEDs and 10, 100, 300 lux for the fluorescent light) to induce browning. The light sources were regulated via a 1 h on/off cycle in a controlled room environment at a temperature of $20^{\circ}C$, 60% humidity, and 1200 ppm $CO_2$ concentration for 60days. The browning effect varied depending on the source and the intensity of illumination. This effect was most effectively induced at $1.5{\mu}mol/m^2s$ for the red and blue LEDs. All light sources induced less browning at the highest intensity of illumination. This indicates that intensity values higher than $20.5{\mu}mol/m^2s$ in the case of the LEDs and 300 lux for the fluorescent light are not effective. After harvesting of the fruit bodies, we measured the weight, length, and width of the pileus and stipe in addition to their chromaticity and hardness. Treatment with $1.5{\mu}mol/m^2s$ blue LED produced the best harvest with the highest average chromaticity, weight (21.2 g), stipe length (30.8 mm), and hardness (377.9 g), with a fine length and width of the pileus.

• Advances in environmental research
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• 제10권1호
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• pp.87-103
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• 2021

Anaerobic digestion (AD) refers to the biological process which can convert organic substrates to biogas in the absence of oxygen. The aim of this study was to determine the capability of feedstock to produce biogas and to quantify the biogas yield from different feedstocks. A co-digestion approach was carried out in a continuous stirred tank reactor operated under mesophilic conditions and at a constant organic loading rate of 0.0756 g COD/ L.day, with a hydraulic retention time of 25 days. For comparison, mono-digestion was also included in the experimental work. 2 L working volumes were used throughout the experimental work. The seed culture was obtained from composting as substrate digestion. When the feedstock was added to seeding, the biogas started to emit after three days of retention time. The highest volume of biogas was observed when the seeding volume used for 1000mL. However, the lowest volume of biogas yield was obtained from both co-digestion reactors, with a value of 340 mL. For methane yield, the highest methane production rate was 0.16 L CH4/mg. The COD with yield was at 8.6% and the lowest was at 0.5%. The highest quantity of methane was obtained from a reactor of Euphorbiaceae peel with added seeding, while the lowest methane yield came from a reactor of Euphorbiaceae stems with added seeding. In this study, sodium bicarbonate (NaHCO3) was used as a buffering solution to correct the pH in the reactor if the reactor condition was found to be in a souring or acidic condition.

• Journal of Microbiology and Biotechnology
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• 제32권6호
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• pp.749-760
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• 2022

α-Galactosidase is a debranching enzyme widely used in the food, feed, paper, and pharmaceuticals industries and plays an important role in hemicellulose degradation. Here, T26, an aerobic bacterial strain with thermostable α-galactosidase activity, was isolated from laboratory-preserved lignocellulolytic microbial consortium TMC7, and identified as Parageobacillus thermoglucosidasius. The α-galactosidase, called T26GAL and derived from the T26 culture supernatant, exhibited a maximum enzyme activity of 0.4976 IU/ml when cultured at 60℃ and 180 rpm for 2 days. Bioinformatics analysis revealed that the α-galactosidase T26GAL belongs to the GH36 family. Subsequently, the pET-26 vector was used for the heterologous expression of the T26 α-galactosidase gene in Escherichia coli BL21 (DE3). The optimum pH for α-galactosidase T26GAL was determined to be 8.0, while the optimum temperature was 60℃. In addition, T26GAL demonstrated a remarkable thermostability with more than 93% enzyme activity, even at a high temperature of 90℃. Furthermore, Ca²⁺ and Mg²⁺ promoted the activity of T26GAL while Zn²⁺ and Cu²⁺ inhibited it. The substrate specificity studies revealed that T26GAL efficiently degraded raffinose, stachyose, and guar gum, but not locust bean gum. This study thus facilitated the discovery of an effective heat-resistant α-galactosidase with potent industrial application. Meanwhile, as part of our research on lignocellulose degradation by a microbial consortium, the present work provides an important basis for encouraging further investigation into this enzyme complex.

• 대한본초학회지
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• 제32권6호
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• pp.9-15
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• 2017

Objectives : Traditional medicines isolated from natural products often have positive effects in the prevention and healing of various immune disorders, such as allergy and atopic inflammation. Scrophulariae Radix (SR) been used in oriental medicine used for treatment of acute and chronic inflammatory diseases. Mast cells are known to play important roles in the initiation of allergic reactions. In this study, we investigated the effects of SR ethanol extract on inflammatory responses in IgE-stimulated RBL-2H3 mast cells. Methods : Rat basophilic leukemia RBL-2H3 cells were purchased from Korean Cell Line Bank (KCLB No. 22256). Cell viability was measured by MTT assay. Assays for ${\beta}-Hexosaminidase$ Secretion : RBL-2H3 cells were sensitized with dinitrophenyl-ImmunoglobulinE (DNP IgE). The next antigen DNP-BSA ($25ng/m{\ell}$) was added for 10 minutes and the reaction was terminated after 5 minutes in the ice bath. To determine ${\beta}-Hexosaminidase$ release, supernatants were aliquoted into 96-well plates. Samples were mixed with substrate solution and incubated for 1 h at $37^{\circ}C$. Absorbance was measured with a spectrophotometer at 405 nm. IL-4 and tumor necrosis $factor-{\alpha}$($TNF-{\alpha}$) concentrations in cell culture supernatants were measured using enzyme-linked immunosorbent assay (ELISA) kits. Results : The cytotoxicity of SRE in RBL-2H3 cells was less than 5%. SRE inhibited DNP-IgE-imduced degranulation of mast cells in RBL-2H3 cells. Also significantly decreased the levels of inflammatory cytokine, IL-4 and TNF-alpha. In this study, the SRE showed potential anti-allergic and antiinflammatory. Conclusions : These results indicate that SRE could be inhibit the allergic response through suppressing the mast cell activation.

• 치위생과학회지
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• 제23권4호
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• pp.369-377
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• 2023

Background: In this study, we investigated the potential use of keratin for oral tissue regeneration. Keratin is well-known for its effectiveness in skin regeneration by promoting keratinization and enhancing the elasticity and activity of fibroblasts. Because of its structural stability, high storability, biocompatibility, and safety in humans, existing research has predominantly focused on its role in skin wound healing. Herein, we propose using keratin proteins as biocompatible materials for dental applications. Methods: To assess the suitability of alpha-keratin protein as a substrate for cell culture, keratin was extracted from human hair via PEGylation. Viabilities of primary human gingival fibroblasts (HGFs) and human oral keratinocytes (HOKs) were assessed. Fluorescence immunostaining and migration assays were conducted using a fluorescence microscope and confocal laser scanning microscope. Wound healing and migration assays were performed using automated software to analyze the experimental readout and gap closure rate. Results: We confirmed the extraction of alpha-keratin and formation of the PEG-g-keratin complex. Treatment of HGFs with keratin protein at a concentration of 5 mg/ml promoted proliferation and maintained cell viability in the test group compared to the control group. HOKs treated with 5 mg/ml keratin exhibited a slight decrease in cell proliferation and activity after 48 hours compared to the untreated group, followed by an increase after 72 hours. Wound healing and migration assays revealed rapid closure of the area covered by HOKs over time following keratin treatment. Additionally, HOKs exhibited changes in cell morphology and increased the expression of the mesenchymal marker vimentin. Conclusion: Our study demonstrated the potential of hair keratin for soft tissue regeneration, with potential future applications in clinical settings for wound healing.

• 한국미생물·생명공학회지
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• 제51권3호
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• pp.257-267
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• 2023

The potential polyhydroxyalkanoates (PHA)-producing bacteria, Bacillus megaterium PP-10, was successfully isolated and studied its feasibility for utilization of pineapple peel waste (PPW) as a cheap carbon substrate. The PPW was pretreated with 1% (v/v) H₂SO₄ under steam sterilization and about 26.4 g/l of total reducing sugar (TRS) in pineapple peel hydrolysate (PPH) was generated and main fermentable sugars were glucose and fructose. A maximum cell growth and PHA concentration of 3.63 ± 0.07 g/l and 1.98 ± 0.09 g/l (about 54.58 ± 2.39%DCW) were received in only 12 h when grown in PPH. Interestingly, PHA productivity and biomass yield (Y_x/s) in PPH was about 4 times and 1.5 times higher than in glucose. To achieve the highest DCW and PHA production, the optimal culture conditions e.g. carbon to nitrogen ratios of 40 mole/mole, incubation temperature at 35℃ and shaking speed of 200 rpm were performed and a maximum DCW up to 4.24 ± 0.04 g/l and PHA concentration of 2.68 ± 0.02 g/l (61% DCW) were obtained. The produced PHA was further examined its monomer composition and found to contain only 3-hydroxybutyrate (3HB). This finding corresponded with the presence of class IV PHA synthase gene. Finally, certain thermal properties of the produced PHA i.e. the melting temperature (T_m) and the glass transition temperature (T_g) were about 176℃ and -4℃, respectively whereas the M_w was about 1.07 KDa ; therefore, the newly isolated B. megaterium PP-10 is a promising bacterial candidate for the efficient conversion of low-cost PPH to PHA.

• 생물환경조절학회지
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• 제18권4호
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• pp.354-362
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• 2009

본 연구는 장미 순환식 수경재배 시 재배시기별로 적합한 배양액을 개발하고자 일본야채다업시험장 표준액을 1/4S, 1/2S, 2/3S, 1S로 하여 펄라이트와 입상 암면을 4:6 부피비로 섞은 고형배지를 이용하여 실험을 실시하였다. 고온기재배의 경우 1/2S 처리구에서, 저온기의 경우 2/3S 처리구에서 광합성 속도, 절화 품질 및 생육이 우수하였다. 이를 토대로 1/2S(고온기), 2/3S(저온기) 처리구의 양수분 흡수율을 기준으로 새로운 배양액을 조성하였다. 이온의 조성은 고온기의 경우 $NO_{3^-}N$ 6.8, $NH_{4^-}N$ 0.7, $PO_{4^-}P$ 2.0, K 3.8, Ca 3.0, Mg 1.2, $SO_{4^-}S$ $1.2me{\cdot}L^{-1}$, 저온기의 경우 $NO_{3^-}N$ 6.8, $NH_{4^-}N$ 0.7, $PO_{4^-}P$ 2.0, K 3.8, Ca 3.0, Mg 1.2, $SO_{4^-}S$ $1.2me{\cdot}L^{-1}$ 이었다. 개발한 배양액의 적합성 평가실험 결과 UOS 배양액은 Ca, P 등의 이온이 장미의 양분흡수율보다 많이 함유된 기존의 배양액과 비교하여 근권 내 EC 변화가 안정적이었다. 또한 절화수량이 재배기간에 관계없이 기존 배양액보다 높은 결과를 나타내었고 특히 저온기 재배시 아이찌현 배양액 처리구에 비하여 수확량이 140% 증가하였다. 따라서 고형배지를 이용한 장미 순환식 수경재배시 새로 개발된 배양액을 사용할 경우 기존 배양액에 비해 비료절감의 효과와 함께 안정적인 생육 및 수량증대를 기대할 수 있다.

• 한국산림과학회지
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• 제41권1호
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• pp.1-6
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• 1979

1. 본시험(本試驗)은 당화기질(糖化基質)로 산오리나무재(材)를 묽은황산(黃酸)으로 전처리(前處理)한 후(後) Trichoderma viride 16374호균(號菌)에서 얻은 cellulase를 작용(作用)시켜 환원당(還元糖)을 생성(生成)할 수 있는 최적조건(最適條件)을 조사(調査)한 것이다. 즉(卽) 산오리나무(10~15년생(年生))의 톱밥을 전처리(前處理)할 때 황산농도(黃酸濃度) 0.3%, 0.6%, 0.9%, 1.2%, 1.5%로 구분(區分)하고 열압시간(熱壓時間)은 $1.5kg/cm^2$에서 15분(分), 30분(分), 45분(分), 60분(分)으로 각각(各各) 구분(區分) 처리(處理)하여 건조(乾燥)시킨것을 다시 $190^{\circ}C$에서 30분간(分間) 열처리후(熱處理後) 60mesh로 분쇄(粉碎)한 것을 당화기질(糖化基質)로 사용(使用)하였다. 그리고 cellulase 반응조건(反應條件)은 0.1M황산완충액(黃酸緩衝液)(pH 5.0) 50ml, 기질량(基質量) 0.5g, 고근효소액(盬析酵素液) 0.5ml를 100ml용(用) 삼각(三角) flask에 넣고 잘 혼합(混合)한 후(後) $40^{\circ}C$에서 96시간(時間) 반응(反應)시켜 이때 생성(生成)된 환원당량(還元糖量)을 조사(調査)하였다. 2. 고근효소액(盬析酵素液)의 조제(調製)는 밀기울액체진탕배양법(液體振盪培養法)에 의(依에) 하여 생성(生成)된 조효소액(粗酵素液)을 유안절포도(硫安飽和度)에 의(依)한 고근효소액(盬析酵素液)을 만들었다. 3. 환원당정량(還元糖定量)은 DNS법(法)에 의(依)하였다. 4. 묽은황산(黃酸)으로 처리(處理)한 톱밥의 환원당(還元糖) 생성량(生成量)은 황산농도(黃酸濃度) 1.5%, $1.5kg/cm^2$에서 열압시간(熱壓時間)은 45분(分)으로 처리(處理)한 것이 16.0%로 최대량(最大量)을 나타냈으나 작용(作用)함으로서 생성(生成)된 환원당량(還元糖量)은 황산농도(黃酸濃度) 0.9%, 열압시간(熱壓時間) 60분(分) 처리(處理)한 것이 23.6%로(본시험(本試驗) 범위(範圍)에서는 47.5% 증가(增加)) 나타났다.

• 생물환경조절학회지
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• 제14권3호
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• pp.203-211
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• 2005

토마토 순환식 고형배지경 시스템에서 생육단계별 작물의 양분 흡수율을 밝히고 최적 배양액을 개발하고자 일본야채시험장 표준액($NO_3-N$ 16.0, $NH_4-N$ 1.3, $PO_4-P$ 4.0, K 8.0, Ca 8.0, Hg 4.0 $me{\cdot}L^{-1}$)을 1/2, 1 및 2배액의 3수준 농도로 조성하여 실험을 수행하였다. 생육초기와 후기 모두 1/2배액 처리에서 생육이 양호하고, 근권 내 pH와 EC의 변화도 적었으며, 식물체내 무기이온 함량도 적정치로 나타났다. 따라서 생육단계별 양${\cdot}$수분 흡수(n/w)율에 의해 개발된 토마토 순환식 배양액 조성은 생육초기에는 $NO_3-N,\;PO_4-p$, K, Ca, Mg 및 $SO_4-S$ 농도가 각각 7.1, 2.1, 4.0, 3.1, 1.2 및 1.2 $me{\cdot}L^{-1}$, 생육후기에는 각각 6.5, 2.3, 3.4, 3.1, 1.1 및 1.1 $me{\cdot}^L^{-1}$로 나타났다. 개발된 배양액의 적합성 검정을 위하여 네델란드 온실작물연구소의 순환식 PTG(Proefstation voor tuinbouw onder glas te Naaldwijk) 배양액으로 비교 실험을 하였다 초기 생육은 PTG와 WU 배양액의 2배액 처리에서, 후기 생육은 WU 2배액 처리에서 가장 좋았다. 정식 후 72일째까지의 수량은 WU 2배액 처리에서 가장 높았으며, 배꼽썩음과는 PTG와 WU 배양액의 모든 농도에서 나타났다. WU 2배액의 EC와 pH변화는 재배되는 동안 안정적이었다. 생육이 높았던 WU 2배액의 식물체내 무기성분 함량 N과 P 함량이 높음에 따라 배양액 조성시 N, P 함량을 낮추어 공급하는 것이 바람직한 것으로 판단된다.

• 한국균학회지
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• 제31권2호
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• pp.59-66
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• 2003

한국산 주걱송편버섯(Pycnoporus cinnabarinus) SCH-3로부터 배지 내로 분비된 laccase를 ultrafiltration과 anion exchange chromatography, adsorption chromatography를 이용하여 분리 정제하고 정제된 호소의 특성을 조사하였다. Laccase는 균주의 일차 대사 과정에서 주로 생산되는 세포외 페놀 산화효소였다. 주걱송편버섯을 기본 배지에서 배양하였을 때 생장은 배양 9일에 최대였고, laccase의 활성은 배양 7일에 최대활성을 나타냈으며 배양액에서 LiP, MnP 그리고 AAO의 활성은 측정되지 않았다. Laccase의 생산에 미치는 유도원의 영향을 조사하기 위하여 배양 중인 주걱 송편버섯에 몇몇 유도원을 첨가한결과, 2,5-xylidine은 대조구에 비하여 laccase의 생산을 25배 증가 시켰다. 정제된 laccase는 SDS 젤 전기영동에서 대략 66 kDa의 분자량을 가지는 단일 폴리펩타이드(single polypeptide)였고, 탄수화물 함량은 9%였다. ABTS [2,2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)]를 기질로 사용하며 정제된 laccase의 $K_{m}}$과$V_{max}$를 조사한 결과 각각 $44.4{\mu}M\;and\;56.0{\mu}mole$로 측정되었다. Laccase 활성의 최적 pH는 3.0이며, 이 효소는 $60{\circ}C$에서 1시간 동안 처리하였을 때 매우 안정적이었고 $80{\circ}C$에서 10분간 처리하였을 때 효소의 활성이 반감되었다. Laccase의 분광학적 특성을 조사한 결과 구리를 포함하는 단백질로 나타났다. 일반적으로 알려진 laccase의 기질들에 대한 특이성을 조사한 결과 5mM ABTS와 5mM hydroquinone에서 높은 활성을 나타내었으며 tyrosine에서는 laccase의 활성이 나타나지 않았다. 저해제의 영향을 조사한 결과, 일반적으로 구리를 포함하는 단백질의 저해제인 $NaN_{3}$, TGA, DDC를 일정농도로 처리한 실험구에서는 효소의 활성이 완전하게 억제되었으며, p-coumaric acid와 EDTA 처리구에서는 효소의 활성이 억제되지 않았다. 한국산 주걱송편버섯 SCH-3 균주로부터 생산되는 laccase의 N-말단의 아미노산의 서열은 P. coccineus의 laccase와 호주에서 분리 동정된 P.cinnabarinus PB의 laccase와 94%가 같았다.